Journal articles on the topic 'LT γδ'
To see the other types of publications on this topic, follow the link: LT γδ.
Create a spot-on reference in APA, MLA, Chicago, Harvard, and other styles
Consult the top 50 journal articles for your research on the topic 'LT γδ.'
Next to every source in the list of references, there is an 'Add to bibliography' button. Press on it, and we will generate automatically the bibliographic reference to the chosen work in the citation style you need: APA, MLA, Harvard, Chicago, Vancouver, etc.
You can also download the full text of the academic publication as pdf and read online its abstract whenever available in the metadata.
Browse journal articles on a wide variety of disciplines and organise your bibliography correctly.
1
Alonso, Sara, Luo Jia, Alyssa Laguerta, and Karen Edelblum. "Expansion of the intraepithelial lymphocyte (IEL) compartment results in an increased bioenergetic profile and reduced IFNγ production in γδ IELs." Journal of Immunology 210, no. 1_Supplement (May 1, 2023): 150.20. http://dx.doi.org/10.4049/jimmunol.210.supp.150.20.
Abstract:
Abstract Intestinal γδ intraepithelial lymphocytes (γδ IEL) serve as the first line of defense against pathogen invasion. We recently identified a transmissible hyperproliferative γδ IEL (γδ HYP) phenotype that protects against enteric infection, and these mice have no signs of overt intestinal pathology. Given that IELs are metabolically constrained to prevent aberrant activation, we hypothesized that the bioenergetics of γδ HYPIELs may regulate their effector function. Electron microscopy revealed a 70% increase in mitochondrial number (p=0.005) and 24% increase in mitochondrial area (p=0.04) in γδ HYPIELs relative to WT. Seahorse mitochondrial stress assays on γδ IELs showed a 50% increase in spare respiratory capacity in γδ HYPIELs compared to WT (p=0.014). Mitochondrial metabolism was shown to influence γδ T cell cytokine production, and similarly, we found that stimulation with PMA and ionomycin reduced the frequency of IFNγ +γδ IELs by 59% in γδ HYPrelative to WT (p<0.0001). We also observed that γδ HYPIELs exhibit higher CD39 expression compared to WT (p<0.0001), and a CD39 hisubset produces less IFNγ compared to CD39 negand CD39 intcells (p<0.0001) The addition of the ATP-synthase inhibitor oligomycin during IEL stimulation increased IFNγ production by both WT and γδ HYPIELs by 66% and 44%, respectively (p<0.0001). Together, our data demonstrate that the shift towards high CD39 expression and elevated mitochondrial metabolism contribute to decreased IFNγ production in γδ HYPIELs. Further understanding of the mechanisms regulating γδ IEL homeostasis and effector function may ultimately allow fine tuning of mucosal surveillance to confer protection against intestinal injury or infection while limiting aberrant cytotoxicity. Supported by NJCCR (COCR23PRF024)
2
Fischer, Matthew, Luo Jia, and Karen Edelblum. "T cell receptor signaling mediates enhanced IFNγ production by γδ intraepithelial lymphocytes in response to type I interferon." Journal of Immunology 210, no. 1_Supplement (May 1, 2023): 150.19. http://dx.doi.org/10.4049/jimmunol.210.supp.150.19.
Abstract:
Abstract Intraepithelial lymphocytes (IEL) expressing the γδ T cell receptor (TCR) provide a rapid response to limit enteric pathogen invasion. Despite constant γδTCR triggering in vivo, these IELs remain immunologically quiescent until their activation threshold is surpassed. Activated γδ IELs limit viral replication by producing type I interferons (IFN), such as IFNα, yet the extent to which IFNα activates γδ IELs remains unknown. To this end, murine small intestinal γδ IELs were stimulated ex vivowith 1 ug/mL αCD3, 10 ng/mL IFNα or both for 5 h. We observed a 26%±10 increase in IFNγ +γδ IELs treated with αCD3 and IFNα compared to αCD3 treatment alone (p<0.0001) and a 2-fold increase in IFNγ MFI (p<0.0001). Using Nur77-GFP mice, we found that only Nur77 +γδ IELs produced IFNγ (p<0.0001), indicating that TCR signaling is necessary for IFNα-induced effector function. Suboptimal αCD3 stimulation (0.1 ug/mL) was sufficient to increase the frequency of IFNγ +γδ IELs following IFNα exposure relative to αCD3 alone (p<0.001). To interrogate the molecular mechanism involved in IFNα co-stimulation, γδ IELs were treated with PMA and/or ionomycin in the presence or absence of IFNα. Ionomycin and IFNα increased the frequency of IFNγ +γδ IELs by 16%±7 compared to ionomycin alone (p<0.0001), whereas PMA had no effect. Pharmacological inhibition of NFAT signaling (10 uM INCA6) abrogated both TCR- or IFNα-induced γδ IEL IFNγ production (p<0.01 or p<0.0001, respectively). Lastly, we observed that the co-stimulatory effect of IFNα was lost in STAT4-deficient γδ IELs compared to WT (p<0.0001). Together, these data indicate that low level TCR signaling through NFAT is required for IFNα to rapidly enhance γδ IEL IFNγ production in a STAT4-dependent manner. This work was supported by the National Institutes of Health R01 DK119349, the New Jersey Commission on Cancer Research, Busch Biomedical Research grant and the RBHS Chancellor Scholar Award (KLE).
3
Hu, Yongxian, Yanjun Gu, Lixia Sheng, Huarui Fu, Kangni Wu, Lifei Zhang, Lizhen Liu, et al. "Decitabine Can Increase the Induction of Regulatory γδ T Cells with Enhanced Immunosuppression on Graft-Versus-Host Disease From Adult Human Peripheral Blood Mononuclear Cells." Blood 118, no. 21 (November 18, 2011): 1901. http://dx.doi.org/10.1182/blood.v118.21.1901.1901.
Abstract:
Abstract Abstract 1901 Regulatory γδ T cells (γδ Tregs) is a novel subset of cells with immunosuppressive function while methods for γδ Treg induction is rarely introduced and its role in graft-versus-host disease (GVHD) prevention remains unkown. Decitabine, a kind of hypomethylating agents, can act synergistically with TGF-β1 to convert a variety of αβ T cells to regulatory αβ T cells with suppressive function but its role in induction and function of γδ Tregs has not been reported. We show here the role of decitabine for the induction of γδ Tregs. Moreover, we provide functional analysis and underlying mechanisms of decitabine-induced γδ Tregs relative to γδ Tregs without decitabine induction as well as in vivo evidences of their preventions on GVHD. Human peripheral blood mononuclear cells (PBMCs) were cultured with IL-2, IL-15, TGF-β1 and zoledronic acid (ZOL). On day 2, 0.5μmol/ml decitabine was added to aliquots of PBMCs. On days 4, 7, and 10, half of the supernatant volume was replaced with media containing cytokines. On day 11, frequencies of γδ Tregs were detected by flow cytometry (FACS). We found the frequency of γδ Tregs was 36.2% in TGF-β1/IL-15/ZOL stimulated group (referred to as common γδ Tregs below) and 59.9% in IL-2/TGF-β1/IL-15/ZOL/decitabine stimulated group (referred to as decitabine-induced γδ Tregs) (p<0.05). In order to compare immunosuppressive function of the two populations, γδ T cells containing γδ Tregs were isolated by magnetic cell sorting system (MACS) and tested for their ability to suppress proliferation of alloreactive PBMCs using CFSE-based assay. After 5 days of in vitro culture, CFSE-labeled PBMCs proliferation was significantly reduced in the presence of enriched γδ Tregs even at 8:1(PBMCs: γδ Tregs) ratio. The inhibition rate was significantly different (decitabine-induced γδ Tregs VS common γδ Tregs at ratio 1:1 is 81.3% VS 68.2%, p<0.05). To clarify the underlying mechanisms we performed ELISA to measure levels of inhibitory cytokines IL-10, IL-4 and TGF-β1 in supernatant of CFSE-based assay. We noted an elevated IL-10 secretion in the decitabine-induced γδ Tregs group compared with common γδ Tregs group (92.7±11pg/ml VS 10.3±2pg/ml at ratio 1:1, p<0.01). We confirmed the result by intracellular IL-10 detection using FACS. Previous reports showed high levels of inducible T-cell costimulator (ICOS) were correlated with IL-10 synthesis. So γδ Tregs were monitored for ICOS expression by FACS. The result revealed that ICOS expression was up-regulated in decitabine-induced γδ Tregs in contrast to common γδ Tregs (MFI: 268 VS 54). Stability of Foxp3 is a critical factor in the immunosuppressive ability of Tregs. Thus we evaluated the frequency of γδ Tregs after 5 days in CFSE-based assay. We observed loss of Foxp3 expression in decitabine-induced γδ Tregs was negligible (<3%) while 15.5% common γδ Tregs lost foxp3 expression. To confirm the results in vitro we tested the functional ability to prevent GVHD in vivo. GVHD was induced in NOD/SCID mice following busulfan and anti-CD122 condition and 1×107 PBMCs transfusion. Animals were co-injected with either decitabine-induced γδ Tregs or common γδ Tregs at a ratio of 1:1. Survival time and GVHD manifestations of the transplanted mice were evaluated. As a result, transplantation of human PBMCs alone induced lethal GVHD with average survival time 25± 8 days while the survival time was 43± 5 days and 58±7 days in mice co-injected with common γδ Tregs and decitabine-induced γδ Tregs, respectively (p<0.05). Clinical manifestations such as hunched back, diarrhea, and body weight loss were statistically different among 3 groups. To investigate the infiltration of human lymphocytes into nonlymphoid tissues in GVHD mice, we performed immunohistochemical analysis of the liver and intestines using anti-human CD45. Remarkably abundant invasion of human CD45+ cells was observed around the veins in the liver and intestines transplanted with PBMCs alone while less invasion in mice co-injected with common γδ Tregs and the lest invasion in mice co-injected with decitabine-induced γδ Tregs. Altogether, our findings reveal that decitabine and the cytokines can efficiently syngenerize to induce γδ Tregs with enhanced immunosuppression on GVHD which are via higher levels of IL-10 production due to ICOS up-regulation as well as stability of Foxp3 expression. Thus γδ Tregs may be potentially exploited therapeutically in a variety of transplantation settings. Disclosures: No relevant conflicts of interest to declare.
4
Fischer, Matthew, Luo Jia, and Karen L. Edelblum. "Type I interferon enhances γδ intraepithelial lymphocyte migratory behavior via CD47 upregulation." Journal of Immunology 206, no. 1_Supplement (May 1, 2021): 17.17. http://dx.doi.org/10.4049/jimmunol.206.supp.17.17.
Abstract:
Abstract γδ intraepithelial lymphocytes (IEL) migrate along the basement membrane and into the lateral intracellular space (LIS) between adjacent intestinal epithelial cells (IEC) and is critical to limit microbial translocation across the barrier. Although activated γδ IELs produce interferon (IFN)-α, whether IFNα directly influences γδ IEL migratory behavior remains unknown. To test this, intravital microscopy was performed on GFP γδ T cell reporter mice treated with PBS or IFNα (1 μg, i.v., 3h). We found that γδ IEL track speed was increased 30% and migration into the LIS was enhanced by 75% in IFNα-treated mice compared to controls. To identify candidate genes involved in regulating γδ IEL motility, we performed RNAseq on γδ IELs isolated from control and IFNα-treated mice. As expected, IFNα induced IFN-stimulated gene expression including CD47 (3.5-fold increase), a transmembrane protein involved in neutrophil transepithelial migration. Stimulation of ex vivo cultured γδ IELs with IFNα resulted in a 3-fold increase in CD47 expression compared to unstimulated controls. Next, to investigate whether IFNα enhances γδ IEL motility in vitro, GFP γδ IELs were co-cultured with enteroids expressing membrane tdTomato and treated with 10ng/ml IFNα for 3h. IFNα increased γδ IEL speed (4.5±0.2 vs 3.4±0.2 μm/min; p<0.05) and displacement (47±3.0 μm vs 34±3.1, p<0.05) relative to untreated controls. Addition of anti-CD47 blocking antibody (MIAP301) abrogated the IFNα-mediated increase in γδ IEL migratory behavior. Taken together, these data demonstrate that IFNα enhances γδ IEL motility via a CD47-mediated mechanism, and suggests that CD47 may represent a conserved mechanism of leukocyte migration within the epithelial barrier.
5
Kimura, Shunsuke, Petri Pölönen, Lindsey Montefiori, Kenneth Caldwell, Ilaria Iacobucci, Chelsey Chen, Anthony Brown, et al. "STAG2/LMO2 Gamma-Delta (γδ) T-ALL: Identification and Characterization of an Extremely High Risk Group of T-ALL in the Very Young." Blood 142, Supplement 1 (November 28, 2023): 845. http://dx.doi.org/10.1182/blood-2023-178688.
Abstract:
Background The prognosis of pediatric T-cell acute lymphoblastic leukemia (T-ALL) has improved with minimal residual disease (MRD)-stratified therapy, however, gamma delta T cell receptor positive (γδ) T-ALL remains a high-risk (HR) group. Limited studies have explored the clinical and genomic characteristics of γδ T-ALL, prompting us to conduct a comprehensive analysis of this entity and to identify determinants of outcome. Methods Through a consortium of 13 groups, we assembled a cohort of 200 patients up to 25 years of age with γδ T-ALL enrolled in clinical trials between 2000 and 2018. Clinical data of patients with non-γδ T-ALL enrolled on the same clinical trials were collected (n = 1,067). Complete remission (CR) was defined when bone marrow (BM) showed M1 cytomorphology and/or MRD <1% without evidence of extramedullary disease at end of induction/consolidation (EOI/EOC) and failure to achieve CR was considered treatment failure. A total of 76 γδ T-ALL samples were analyzed by whole genome (WGS) and/or transcriptome (RNAseq) sequencing. Results The frequency of γδ T-ALL was 8.0% of T-ALL cases. Patients with γδ T-ALL exhibited a higher rate of poor prednisone response ( P<0.01), MRD >1% at day 15 ( P<0.01), at EOI ( P<0.01) and EOC ( P<0.01), compared to non-γδ T-ALL cases. Furthermore, patients with γδ T-ALL showed significantly worse 5-year event free survival (EFS, 65% v. 78%, P<0.01) and overall survival (OS, 77% vs 83%, P=0.048). Almost all relapses of γδ T-ALL were isolated BM, while the central nervous system was the main site of relapse in non-γδ T-ALL, suggesting slow treatment response and chemo-resistance to the current treatment in γδ T-ALL. However, γδ T-ALL showed a higher rate of toxic death during treatment (7.6% vs 4.0%, P<0.01), suggesting the need for different therapeutic strategies and risk-classification, rather than treatment intensification. Strikingly, patients less than 3 years of age with γδ T-ALL exhibited significantly poor EFS (33% v. 70% [3-10 years] and 73% [>10], P<0.01) and OS (49% v. 78% [3-10] and 82% [>10], P< 0.01) ( Fig. A), a difference not observed in non-γδ T-ALL. MRD >1% at EOI showed poor EFS (51% v. 96% [MRD<0.01%] and 91% [1%>MRD>0.01%], P<0.01) and OS (66%). Integrated analysis of WGS and RNAseq identified enrichment of several genomic subtypes in γδ T-ALL, including STAG2/LMO2, hyperdiploidy with recurrent gains of chromosomes 8, 10, 11, 13q and 19, a recently identified “LMO2 γδ-like” subtype with distinct gene expression and LMO2/MYC/MYCN alterations, TLX3-rearranged (-R), and PICALM::MLLT10. No TAL1 nor TLX1-R were detected. STAG2/LMO2 was associated with age at diagnosis before 3 years, and extremely poor outcome, with 4 out of 5 cases dying within three years of diagnosis ( Fig. B). Of 24 STAG2/LMO2 T-ALL (additional 5 non-γδ, 13 TCR unknown cases), 22 of which were diagnosed by age three. All STAG2/LMO2 cases had alterations resulting in LMO2 activation and STAG2 inactivation, most commonly a single rearrangement between these two genes, and upregulation of HBE1, the LIN28-let7 pathway and stem cell proliferation pathways, suggesting a fetal hematopoietic origin. STAG2 has a critical role in the maintenance of enhancer-promoter looping mediated by the cohesin complex. To examine the consequences of STAG2 alterations, we performed integrated genomic/epigenomic analysis of the STAG2/LMO2 (MOLT-14 and PER-117) and STAG2 knockout (KO)/addback T-ALL lines. Chromatin loop sizes defined by H3K27ac HiChIP was highest in STAG2/LMO2 lines compared to other T-ALL. Following restoration of STAG2 expression in MOLT-14, CD34 and ID1/2 were down-regulated and H3K27ac was enriched in pathways related to T-cell differentiation. STAG2 KO in the non- STAG2/LMO2, LMO2-activated line PF382 identified genes also upregulated in STAG2/LMO2 primary samples, including CDK4 and STAG1. STAG2 KO lines exhibited partial compensation of STAG2 binding sites by STAG1 and upregulation of γδ-related genes, RORC and ID1/3. High throughput screening of 2,050 small molecules identified efficacy of HDAC, CDK and PARP inhibitors in STAG2/LMO2 lines. Conclusion Very young onset γδ T-ALL, but not non-γδ T-ALL, is enriched for the STAG2/LMO2 subtype and is a very high risk form of T-ALL. STAG2 loss perturbs chromatin organization and hematopoietic differentiation. Moreover, we demonstrate efficacy of novel therapeutic approaches that are needed to cure this form of leukemia.
6
Liang, Shuang, Jiangying Liu, Haitao Gao, Ruoyang Liu, Ning Wu, Tianhui Dong, and Xiaojun Huang. "Induced CD25+CD127dim Γδ Tregs in Acute Myeloid Leukemia Suppress the Activity of Normal Αβ T Cells." Blood 136, Supplement 1 (November 5, 2020): 27–28. http://dx.doi.org/10.1182/blood-2020-136541.
Abstract:
Acute myeloid leukemia (AML) is an aggressive hematologic malignancy in adults with dismal outcomes. γδ T cells have been reported to exert effective anti-tumor activities on various types of solid tumors and hematologic malignancies, and thus become a potential weapon used in the treatment of cancer. However, compared to the striking results demonstrated in vitro and in mouse models, the clinical efficacy of γδ T cells-based immunotherapy is disappointing. Meanwhile, recent studies identified a pro-tumor role of γδ T cells in some types of solid tumors. Whether γδ T cells could be transformed from warriors to foe in the environment of leukemia is unclear. In the current study, we aim to dissect the abnormal changes and functions of intrinsic γδ T cells in the context of AML. Considering bone marrows (BM) are the primary tumor foci of AML and likely provide more representative immune characteristics during leukemia development, the immunophenotyping analyses of γδ T cells in BM from newly diagnosed AML patients (n = 21) were detected using flow cytometry, compared with healthy individual controls (n = 21). We observed that the proportion of γδ T cells and the composition of Vδ1+ and Vδ2+ subgroups were comparable between AML patients and healthy controls. Interestingly, a dramatically elevated subpopulation in γδ T cells expressing CD25 and dim CD127 was observed in AML patients compared with that in healthy controls (3.2% versus 0.8%, P < 0.001). Since their phenotypes are akin to the conventional CD4+ regulatory T cells (Tregs), these CD25+CD127dim γδ T cells are called γδ Tregs here. Paired comparison was also performed on AML patients at diagnosis (n = 13) and completed remission (CR) after chemotherapy (n = 13). A significant decrease in proportion of γδ Tregs was observed at CR, compared with that at newly diagnoses (3.1% versus 0.4% (P < 0.001). These results suggest that this aberrant γδ T subset, γδ Tregs, is induced in the microenvironment of AML and can be reversed after effective therapy for elimination of leukemia blasts. To further explore the direct effect of AML blasts on γδ T cells, primary malignant cells were sorted from AML patients, and co-cultured with peripheral blood mononuclear cells (PBMCs) isolated from healthy donors. As expected, γδ Tregs was significantly induced and the frequency was continuously elevated from 0.2% to 4.0% following the increased ratios of AML cells (P < 0.001). These results confirmed that AML cells were capable to trigger the transformation of normal γδ T cells into CD25+CD127dim γδ Tregs. To determine the function of this special γδ T subset, CD25+CD127dim γδ Tregs were expanded in vitro and co-cultured with AML cell lines, U937 cells. Unlike pamidronate-expanded cytotoxic Vδ2+ T cells, γδ Tregs lost the ability of killing AML cells. Furthermore, we explored the impact of γδ Tregs on the anti-AML activity of normal αβ T cells. γδ T cell-depleted PBMCs (more than 90% are αβ T cells) of healthy donors were co-cultured with U937 cells, with or without autologous γδ Tregs. Flow cytometry analysis showed that 7-AAD-positive fraction in U937 cells was dramatically increased from 0.3% to 7.6% after co-culture with PBMCs and without γδ Tregs. Whereas the mortality of U937 cells was gradually declined (from 7.6% to 4%) in the presence of γδ Tregs at increasing ratios (P = 0.031). During this process, γδ Tregs remarkably downregulated the expressions of HLA-DR and CD38, and productions of TNF-α and IFN-γ in both CD4+ and CD8+ αβ T cells. These findings indicated that γδ Tregs significantly impaired the cytotoxic ability of effector αβ T cells on AML cells, suggesting γδ Tregs play a suppressive role in the anti-leukemia immunity. In summary, we first reported the significant induction of CD25+CD127dim γδ Tregs in the primary BM of AML patients. These γδ Tregs did not exert anti-leukemia activity and attenuated the cytotoxic effect of normal αβ T cells. Our findings will help advance the current understanding of immune mechanism associated with leukemia cell evasion. It also proposes that boosting the cytotoxic γδ T cells should be coupled with inhibition of the suppressive γδ Tregs, for future improvement of γδ T-cell-based immunotherapy. Disclosures No relevant conflicts of interest to declare.
7
Silva, Polyana Barbosa, Márcia Antoniazi Michelin, Millena Prata Jammal, and Eddie Fernando Cândido Murta. "Immunological Characteristics between αβ TDC and γδ TDC Cells in the Spleen of Breast Cancer-Induced Mice." Revista Brasileira de Ginecologia e Obstetrícia / RBGO Gynecology and Obstetrics 43, no. 05 (May 2021): 368–73. http://dx.doi.org/10.1055/s-0041-1730286.
Abstract:
Abstract Objective To evaluate the antitumoral role of γδ TDC cells and αβ TDC cells in an experimental model of breast cancer. Methods Thirty female Balb/c mice were divided into 2 groups: control group (n = 15) and induced-4T1 group (n = 15), in which the mice received 2 × 105 4T1 mammary tumor cell line. Following the 28-day experimental period, immune cells were collected from the spleen and analyzed by flow cytometry for comparison of αβ TDC (TCRαβ+ CD11c+MHCII+) and γδ TDC (TCRγδ+CD11c+MHCII+) cells regarding surface markers (CD4+ and C8+) and cytokines (IFN-γ, TNF-α, IL-12 and IL-17). Results A total of 26.53% of γδ TDC - control group (p < 0.0001) - the proportion of αβ TDC was lower in splenic cells than γδ TDC; however, these 2 cell types were reduced in tumor conditions (p < 0.0001), and the proportion of IFN-γ, TNF-α, IL-12 and IL-17 cytokines produced by γδ TDC was higher than those produced by αβ TDC, but it decreased under conditions of tumor-related immune system response (p < 0.0001). Conclusion Healthy mice engrafted with malignant cells 4T1 breast tumor presented TDC with γδ TCR repertoire. These cells express cytotoxic molecules of lymphocytes T, producing anti-tumor proinflammatory cytokines.
8
Maeda, Yoshinobu, Pavan Reddy, Kathleen P. Lowler, Chen Liu, Dennis Keith Bishop, and James L. M. Ferrara. "Critical role of host γδ T cells in experimental acute graft-versus-host disease." Blood 106, no. 2 (July 15, 2005): 749–55. http://dx.doi.org/10.1182/blood-2004-10-4087.
Abstract:
Abstract γδ T cells localize to target tissues of graft-versus-host disease (GVHD) and therefore we investigated the role of host γδ T cells in the pathogenesis of acute GVHD in several well-characterized allogeneic bone marrow transplantation (BMT) models. Depletion of host γδ T cells in wild-type (wt) B6 recipients by administration of anti-T-cell receptor (TCR) γδ monoclonal antibody reduced GVHD, and γδ T-cell-deficient (γδ-/-) BM transplant recipients experienced markedly improved survival compared with normal controls (63% vs 10%, P < .001). γδ T cells were responsible for this difference because reconstitution of γδ-/- recipients with γδ T cells restored GVHD mortality. γδ-/- recipients showed decreased serum levels of tumor necrosis factor α (TNF-α), less GVHD histopathologic damage, and reduced donor T-cell expansion. Mechanistic analysis of this phenomenon demonstrated that dendritic cells (DCs) from γδ-/- recipients exhibited less allostimulatory capacity compared to wt DCs after irradiation. Normal DCs derived from BM caused greater allogeneic T-cell proliferation when cocultured with γδ T cells than DCs cocultured with medium alone. This enhancement did not depend on interferon γ (IFN-γ), TNF-α, or CD40 ligand but did depend on cell-to-cell contact. These data demonstrated that the host γδ T cells exacerbate GVHD by enhancing the allostimulatory capacity of host antigen-presenting cells. (Blood. 2005;106:749-755)
9
Boissière-Michot, Florence, Ghita Chabab, Caroline Mollevi, Séverine Guiu, Evelyne Lopez-Crapez, Jeanne Ramos, Nathalie Bonnefoy, Virginie Lafont, and William Jacot. "Clinicopathological Correlates of γδ T Cell Infiltration in Triple-Negative Breast Cancer." Cancers 13, no. 4 (February 12, 2021): 765. http://dx.doi.org/10.3390/cancers13040765.
Abstract:
The prognostic impact of the different tumor-infiltrating lymphocyte (TIL) subpopulations in solid cancers is still debated. Here, we investigated the clinicopathological correlates and prognostic impact of TILs, particularly of γδ T cells, in 162 patients with triple-negative breast cancer (TNBC). A high γδ T cell density (>6.625 γδ T cells/mm2) was associated with younger age (p = 0.008), higher tumor histological grade (p = 0.002), adjuvant chemotherapy (p = 0.010), BRCA1 promoter methylation (p = 0.010), TIL density (p < 0.001), and PD-L1 (p < 0.001) and PD-1 expression (p = 0.040). In multivariate analyses, γδ T cell infiltration (cutoff = 6.625 γδ T cells/mm2) was an independent prognostic factor (5-year relapse-free survival: 63.3% vs. 89.8%, p = 0.027; 5-year overall survival: 73.8% vs. 89.9%, p = 0.031, for low vs. high infiltration). This prognostic impact varied according to the tumor PIK3CA mutational status. High γδ T cell infiltration was associated with better survival in patients with PIK3CA wild-type tumors, but the difference was not significant in the subgroup with PIK3CA-mutated tumors. Altogether, these data suggest that high γδ T cell infiltrate is correlated with immune infiltration and might represent a candidate prognostic tool in patients with TNBC.
10
Tuengel, Jessica, Sanya Ranchal, Alexandra Maslova, Gurpreet Aulakh, Maria Papadopoulou, Sibyl Drissler, Bing Cai, et al. "Characterization of Adaptive-like γδ T Cells in Ugandan Infants during Primary Cytomegalovirus Infection." Viruses 13, no. 10 (October 3, 2021): 1987. http://dx.doi.org/10.3390/v13101987.
Abstract:
Gamma-delta (γδ) T cells are unconventional T cells that help control cytomegalovirus (CMV) infection in adults. γδ T cells develop early in gestation, and a fetal public γδ T cell receptor (TCR) clonotype is detected in congenital CMV infections. However, age-dependent γδ T cell responses to primary CMV infection are not well-understood. Flow cytometry and TCR sequencing was used to comprehensively characterize γδ T cell responses to CMV infection in a cohort of 32 infants followed prospectively from birth. Peripheral blood γδ T cell frequencies increased during infancy, and were higher among CMV-infected infants relative to uninfected. Clustering analyses revealed associations between CMV infection and activation marker expression on adaptive-like Vδ1 and Vδ3, but not innate-like Vγ9Vδ2 γδ T cell subsets. Frequencies of NKG2C+CD57+ γδ T cells were temporally associated with the quantity of CMV shed in saliva by infants with primary infection. The public γδ TCR clonotype was only detected in CMV-infected infants <120 days old and at lower frequencies than previously described in fetal infections. Our findings support the notion that CMV infection drives age-dependent expansions of specific γδ T cell populations, and provide insight for novel strategies to prevent CMV transmission and disease.
11
Golovchenko, Natasha B., Weili Xu, Andrew Fong, Lanjing Zhang, and Karen Edelblum. "Reduced γδ intraepithelial lymphocyte number and motility precede the onset of Crohn’s disease-like ileitis." Journal of Immunology 210, no. 1_Supplement (May 1, 2023): 150.12. http://dx.doi.org/10.4049/jimmunol.210.supp.150.12.
Abstract:
Abstract Crohn’s disease (CD) is a relapsing, remitting disease resulting in chronic intestinal inflammation. Recent studies of resected ileal tissue from CD patients reported a loss of intraepithelial lymphocytes expressing the γδ T cell receptor (γδ IELs). γδ IELs continuously survey the epithelial barrier and protect against experimental colitis; however, the role of γδ IELs in the initiation of ileitis is unknown. To this end, we profiled the IEL compartment in TNF ΔARE/+mice, which develop chronic CD-like ileitis at 8 wks old, prior to and during ileitis onset (4–10 wks). We observed a 2.5-fold reduction in the frequency of ileal γδ IELs in 5-wk-old TNF ΔARE/+mice compared to TNF +/+(WT) littermates (p<0.001). Using intravital microscopy, we observed reduced γδ IEL surveillance behavior (10.6 vs 6.4 mm/min track speed mean, p<0.0001) in TNF ΔARE/+mice compared to WT at 6 wks. Analysis of cleared ileal tissue from 6-wk-old WT and TNF ΔARE/+mice revealed a reduction in the number of γδ IELs in villus tips in TNF ΔARE/+mice relative to WT, thereby increasing the proportion of γδ IELs found in the crypt region. Based on several reported links between γδ IELs and Paneth cells (PC), we quantified PCs by lysozyme immunostaining in ileal tissue. We found an increased proportion of morphologically abnormal PCs in 6-wk-old TNF ΔARE/+mice, reflecting PC dysfunction consistent with CD pathology. These findings are supported by a reduction in ileal Lyz1 expression in TNF ΔARE/+mice compared to WT prior to ileitis onset (p=0.02). Together, these studies demonstrate that γδ IELs exhibit impaired surveillance behavior and altered distribution along the crypt/villus axis prior to ileitis onset, which coincides with PC dysfunction in CD-like ileitis. COCR23PRF017 NJCCR Predoctoral Fellowship Golovchenko (PI) June 1, 2022 – May 31, 2024 γδ IEL surveillance behavior in the onset of chronic ileitis
12
Lu, Hai-Yan, Tian-Shu Peng, Xiang-Dang Hu, Shuai-Jun Li, Min Luo, Yong-Heng He, and Tian Nie. "Quercetin potentiates the effect of γδ T cells via modulating the expressions of Granzyme B, perforin and IFN-γ and also regulates the Wnt/β-catenin signalling pathway in human colon cancer cells." Bangladesh Journal of Pharmacology 10, no. 2 (April 1, 2015): 251. http://dx.doi.org/10.3329/bjp.v10i2.20387.
Abstract:
<p>Cancer accounts as one of the leading causes of morbidity and mortality. Recent studies focus on the efficiency of phytochemicals in cancer therapy. Influence of quercetin, a flavonoid on the effect of γδ T cells and Wnt/β-catenin signalling pathway in human colon cancer cells (HT55 and HCT116) was investigated. Quercetin at 15-120 µM was observed to markedly reduce the viability of HT55 and HCT116 cells. Quercetin exposure significantly increased γδ T cell proliferation and also raised the expressions of granzyme B (Gra B), perforin (PFP), and interferon- γ (IFN-γ) in γδ T cells. Reduced β-catenin expression with increased expressions of phosphorylated- β-catenin, axin1 and 2 were observed in HT55 and HCT116 cells on exposure to quercetin. However β-actin expression was found to be not much altered. The results suggest that quercetin was able to efficiently potentiate the effect of γδ T cells and modulate Wnt/β-catenin signalling pathway.</p><p> </p>
13
Andreu-Ballester, Juan Carlos, Lorena Galindo-Regal, Julia Hidalgo-Coloma, Carmen Cuéllar, Carlos García-Ballesteros, Carolina Hurtado, Natalia Uribe, et al. "Differences in circulating γδ T cells in patients with primary colon cancer and relation with prognostic factors." PLOS ONE 15, no. 12 (December 16, 2020): e0243545. http://dx.doi.org/10.1371/journal.pone.0243545.
Abstract:
Downregulation of the T cell system has been proposed as a mechanism to block immunity in colonic cancer (CC). However, little has been studied about circulating αβ and γδ T cells and their immunological status in newly diagnosed patients. The aim of this study was to characterize the αβ and γδ T cell subsets in peripheral blood of patients with CC matched with healthy volunteers. In this prospective case-control study, blood samples were obtained from 96 patients with newly diagnosed treatment-naïve infiltrating colonic adenocarcinoma and 48 healthy volunteers. Pathological report at surgery was obtained from all CC patients. A significant decrease in CD3+ γδ T cells and CD3+CD8+ γδ T cells (p<0.001) were observed in CC patients. Apoptosis was significantly increased in all conventional and both αβ and γδ T cell subsets in patients with CC vs healthy subjects. γδ T cells were decreased in peripheral blood of patients with microscopic infiltration in tissues, history of cancer and synchronous colon cancer (p < 0.05). IFN-γ was significantly reduced in CC patients compared to controls. Cytotoxic effector γδ T cells TEMRA (CD8 and CD56) are the proportionally most abundant T cells in peripheral blood of CC patients. Patients with CC present a deep downregulation in the systemic T-cell immunity. These variations are evident through all tumor stages and suggest that a deficiency in γδ T cell populations could be preventing control of tumor progression. This fact prove the role of immunomodulation on CC carcinogenesis.
14
Pawlik-Gwozdecka, Dorota, Justyna Sakowska, Maciej Zieliński, Magdalena Górska-Ponikowska, Francesco Cappello, Piotr Trzonkowski, and Maciej Niedźwiecki. "Association between Serum Heat Shock Proteins and Gamma-Delta T Cells—An Outdated Clue or a New Direction in Searching for an Anticancer Strategy? A Short Report." Applied Sciences 11, no. 16 (August 9, 2021): 7325. http://dx.doi.org/10.3390/app11167325.
Abstract:
HSPs demonstrate a strong association with gamma-delta (γδ) T cells. Most of the studies regarding interactions between the parameters were conducted in the 1990s. Despite promising results, the concept of targeting γδ T cells by HSPs seems to be a forgotten direction due to potent non-peptidic phosphoantigens rather than HSPs have been found to be the essential stimulatory components for human γδ cells. Currently, with greater knowledge of lymphocyte diversity, and more accurate diagnostic methods, we decided to study the correlation once again in the neoplastic condition. Twenty-one children with newly diagnosed acute lymphoblastic leukaemia (ALL) were enrolled on the study. Serum HSP90 concentrations were evaluated by an enzyme-linked immunosorbent assay (ELISA), subsets of γδ T cells (CD3+ γδ, CD3+ γδ HLA/DR+, CD4+ γδ and CD8+ γδ) by flow cytometry. We have shown statistically relevant correlations between serum HSP90 and CD3+ HLA/DR+ γδ T cells in paediatric ALL at diagnosis (R = 0.53, p < 0.05), but not after induction chemotherapy. We also have demonstrated decreased levels of both serum HSP90 and CD3+ HLA/DR+ γδ T cells before treatment, which may indirectly indicate dose-dependent unknown interaction between the parameters. The results of our study may be a good introduction to research on the association between HSPs and CD3+ HLA/DR+ γδ T cells, which could be an interesting direction for the development of anti-cancer strategies, not just for childhood ALL.
15
Gaballa, Ahmed, Lucas C. M. Arruda, Emelie Rådestad, and Michael Uhlin. "CD8+γδ T Cells Are More Frequent in CMV Seropositive Bone Marrow Grafts and Display Phenotype of an Adaptive Immune Response." Stem Cells International 2019 (December 6, 2019): 1–13. http://dx.doi.org/10.1155/2019/6348060.
Abstract:
The role of gamma delta (γδ) T cells in human cytomegalovirus (HCMV) immune surveillance has been the focus of research interest for years. Recent reports have shown a substantial clonal proliferation of γδ T cells in response to HCMV, shedding light on the adaptive immune response of γδ T cells. Nevertheless, most efforts have focused on Vδ2negγδ T cell subset while less attention has been given to investigate other less common γδ T cell subsets. In this regard, a distinct subpopulation of γδ T cells that expresses the CD8 coreceptor (CD8+γδ T cells) has not been thoroughly explored. Whether it is implicated in HCMV response and its ability to generate adaptive response has not been thoroughly investigated. In this study, we combined flow cytometry and immune sequencing of the TCR γ-chain (TRG) to analyze in-depth bone marrow (BM) graft γδ T cells from CMV seropositive (CMV+) and CMV seronegative (CMV-) donors. We showed that the frequency of CD8+γδ T cells was significantly higher in CMV+ grafts compared to CMV- grafts (P<0.001). Further characterization revealed that CD8+γδ T cells from CMV+ grafts express Vγ9- and preferentially differentiated from a naive to terminal effector memory phenotype (CD27low/-CD45RO-). In line with these findings, TRG immune sequencing revealed clonal focusing and reduced usage of the Vγ9/JP gene segment in a CMV+ graft. Furthermore, CD8+γδ T cells showed an enhanced response to TCR/CD3 and cytokine stimulation in contrast to CD8-γδ T cells. We conclude that γδ T cells in BM grafts are reshaped by donor CMV serostatus and highlight the potential adaptive role of CD8+γδ T cells in HCMV immune response.
16
Das, Dayasagar, Vivek Anand, Parul Singh Antil, Sujay Khandpur, V. K. Sharma, and Alpana Sharma. "Involvement of γδ T cells and scavenger receptors in the immunopathogenesis of Pemphigus Vulgaris (BA11P.129)." Journal of Immunology 194, no. 1_Supplement (May 1, 2015): 184.11. http://dx.doi.org/10.4049/jimmunol.194.supp.184.11.
Abstract:
Abstract Pemphigus Vulgaris (PV) is an auto-immune blistering disease mostly due to formation of auto-antibodies against desmoglein3 (Dsg3) and aberrant T cell function. γδ T cells play significant role in the immune surveillance at the epithelial surface and modulate other immune cells. Scavenger receptors (CD36 and CD163) are Pattern recognizing receptor (PRR) molecules and key players in several inflammatory diseases. γδ T cells and its signature molecular markers has found to be involved in several autoimmune diseases. However the role of γδ T cells and associated Scavenger receptor molecules are not characterized in the Immunopathogenesis of PV. In our study we observed significantly higher frequency of γδ T cells (6.7% vs. 3.8%) and IL17 producing γδ T cells in circulation of PV patients (n=25) vs. controls (n=15) respectively along with Th1 polarization (p<0.010) suggesting the potential role of γδ T cells and IL17 secreting γδ T cells in the pathogenesis of PV. Cytotoxic activity of γδ T cells from PV patients (26±11%) was observed to be higher as compare to control (12±7%). The molecular expression (mRNA) of scavenger receptors CD36 and CD163 were elevated in patients as compared to controls indicating the possible involvement of these PRR along with γδ T cells. These findings may help to understand the pathogenesis of PV and find better approach for novel therapeutics.
17
Dong, Ruoyu, Yixi Zhang, He Huang, Xun Zeng, and Haowen Xiao. "A Novel Strategy to Produce CAR-Γδ T Cells Via Targeted Knockout By CRISPR/Cas9 and Targeted Knockin By AAV." Blood 142, Supplement 1 (November 28, 2023): 6840. http://dx.doi.org/10.1182/blood-2023-189179.
Abstract:
Chimeric antigen receptor (CAR)-γδ T cells are promising in immunotherapy owing to the dual recognition system of γδ T cells, which is potential to exhibit stronger anti-tumor effect. Up to date, established CAR-γδ T cells exhibit poor anti-tumor efficiency and are commonly generated using lentiviral (LV) or gamma-retroviral (γRV) vectors, which may increase the risk of insertional mutagenesis. Nowadays, clustered regularly interspaced palindromic repeats/CRISPR-associated protein 9 (CRISPR/Cas9) has been widely used to cleave the genomic sequence precisely, as well as Adeno-associated Virus (AAV) being used to deliver DNA templates for target sequence knock-in (KI) mediated by homology-directed repair, which could contribute to achieve the accurate insertion of CAR at a specific site. Furthermore, the endogenous promoter of TCR has been used to express CAR molecules with stable expression and better antitumor function in the production of CAR-αβ T cells. In the present study, we established and optimized the strategy to generate CAR-γδ T cells, combining the site-specific knock-out (KO) targeting the TCR delta chain constant region (TRDC) site of γδ T cells using CRISPR/Cas9 and site-specific CAR knock-in using AAV. Firstly, we screened out a functional guide RNA (gRNA) targeting the γδ TCR TRDC locus by prediction tools and validation in cell line model experiments, and achieved an INDEL frequency of 31.8%, which showed a high activity of gRNA. Then, we optimized the option to increase KO efficiency including exploring appropriate Cas9 amount, ratio of gRNA to Cas9 and cell number titration. Besides, we designed AAV vector carrying CAR sequence targeting the gRNA aiming site selected above (Figure1A). After that, we used CRISPR/Cas9 and AAV to knock out TCR and knock in CAR sequence in γδ T cells. We tried different electroporation procedures (X001, T007, U014, T023) and cell culture conditions (cultured in medium containing zoledronic acid (ZOL)/ IL2 for 5 days or for 10 days) to select out the optimal procedure to acquire high KO/KI efficiency and high cell proliferation rate. When we used PBMC being cultured in medium containing zoledronic acid (ZOL)/ IL2 for 5 days (Day 5) to generate CAR-γδ T cells, cells electroporated using X001 procedure showed the best TCR KO efficiency of ~80%（p<0.05）and CAR expression of ~20% (p< 0.01), as well as keeping the highest cell proliferation rate than other procedures (p<0.0001) (Figure 1B and 1C). If we used PBMC being cultured in medium containing zoledronic acid (ZOL)/ IL2 for 10 days (Day 10) to generate CAR-γδ T cells, U014 program also exhibited a relative high CAR expression and the significant quicker cell growth than others in the later period (p<0.01) (Figure 1B and 1C). However, the later proliferation rate of CAR-γδ T cells generated at Day 5 was superior to that generated at Day10. Thus, we determined that a 5 days' cell culture and the use of X001 program were the optimal condition to generate CAR-γδ T cells. Besides, we found that KI efficiency is positively correlative with Multiplicity of infection (MOI). While MOI comes to 1.2×10 5, the KI efficiency couldn't be increased (Figure 1D). Finally, we evaluated the anti-tumor function of CAR-γδ T cells . Antigen stimulated CAR-γδ T cells significantly released TNFα and INFγ, but failed to produce IL2 (Figure 1E). Besides, CAR-γδ T cells could significantly lysis different tumor cells even at a low effector/target ratio (E/T) (Figure 1G). Interestingly, the proportion of memory phenotype (CD62L + ) in CAR-γδ T cells after being stimulated was comparable to that of CAR-γδ T cells without stimulation, indicating the potential of the great anti-tumor activity of CAR-γδ T cells after being stimulated in tumor environment (Figure 1F). Our new method successfully generated CAR-γδ T cells with a CAR expression of 20% and in a high proliferation rate, which was determined to exert effective anti-tumor effects.
18
Arruda, Lucas C. M., Ahmed Gaballa, and Michael Uhlin. "Impact of γδ T cells on clinical outcome of hematopoietic stem cell transplantation: systematic review and meta-analysis." Blood Advances 3, no. 21 (November 12, 2019): 3436–48. http://dx.doi.org/10.1182/bloodadvances.2019000682.
Abstract:
Abstract Allogeneic hematopoietic stem cell transplantation (HSCT) using αβ T-/B-cell–depleted grafts recently emerged as a transplant strategy and highlighted the potential role of γδ T cells on HSCT outcomes. Our aim was to scrutinize available evidence of γδ T-cell impact on relapse, infections, survival, and acute graft-versus-host disease (aGVHD). We performed a systematic review and meta-analysis of studies assessing γδ T cells in HSCT. We searched PubMed, Web of Science, Scopus, and conference abstracts from inception to March 2019 for relevant studies. We included all studies that assessed γδ T cells associated with HSCT. Data were extracted independently by 2 investigators based on strict selection criteria. A random-effects model was used to pool outcomes across studies. Primary outcome was disease relapse. We also assessed infections, survival, and aGVHD incidence. The review was registered with PROSPERO (CRD42019133344). Our search returned 2412 studies, of which 11 (919 patients) were eligible for meta-analysis. Median follow-up was 30 months (interquartile range, 22-32). High γδ T-cell values after HSCT were associated with less disease relapse (risk ratio [RR], 0.58; 95% confidence interval [95% CI], 0.40-0.84; P = .004; I2 = 0%), fewer viral infections (RR, 0.59; 95% CI, 0.43-0.82; P = .002; I2 = 0%) and higher overall (HR, 0.28; 95% CI, 0.18-0.44; P < .00001; I2 = 0%) and disease-free survivals (HR 0.29; 95% CI, 0.18-0.48; P < .00001; I2 = 0%). We found no association between high γδ T-cell values and aGVHD incidence (RR, 0.72; 95% CI, 0.41-1.27; P = .26; I2 = 0%). In conclusion, high γδ T cells after HSCT is associated with a favorable clinical outcome but not with aGVHD development, suggesting that γδ T cells have a significant effect on the success of HSCT. This study was registered with PROSPERO as #CRD42019133344.
19
Wu, Xiuli, Li Xuan, Xu Wang, Sijian Yu, Zhengshan Yi, Min Dai, Yu Zhang, Zhongxin Zheng, Qifa Liu, and Yangqiu Li. "Enhancement Of Human Regulatory γδ T Cells In Vitro Induced By Granulocyte Colony-Stimulating Factor." Blood 122, no. 21 (November 15, 2013): 5418. http://dx.doi.org/10.1182/blood.v122.21.5418.5418.
Abstract:
Abstract Background and Objective Graft versus host disease (GVHD) is the main complications following allogeneic hematopoietic stem cell transplantation (allo-HSCT). Regulatory γδ T cells (γδ Tregs), which express Foxp3 and primarily belong to CD27+CD25high phenotype, are a novel subset of cells with immunosuppressive function and have the potential application prospect in GVHD therapy. Peripheral blood mononuclear cells (PBMCs) could be induced to generate γδ Tregs in vitro by stimulating with anti-TCR γδ and transforming growth factor-β (TGF-β). Our previous studies demonstrated that granulocyte colony-stimulating factor (G-CSF) had immunomodulatory effect on T cells; G-CSF mobilization influenced the distribution and clonality of TRGV and TRDV repertoire (T cell receptors of γδ T cells), and significant positive correlation was observed between the invariable clonality of TRDV1 gene repertoire after G-CSF mobilization and low incidence of GVHD in recipients. To further characterize the immunoregulatory functions of G-CSF, this study explored the possibility of γδ Tregs induced by G-CSF in vitro. Methods PBMCs of healthy donors were cultured in vitro by stimulating with anti-TCR γδ and different cytokines for 9-12 days. The culture system was grouped based on the difference of added cytokines, including: (1) TGF-β (2) G-CSF (3) TGF-β+G-CSF (4) blank group. The induced cells were used as effector cells in the carboxylfluorescein diacetate succinimidyl ester (CFSE) assays and cell immunophenotyping was analyzed by flow cytometry. Autologous CD4+T cells were purified by microbeads, labeled with CFSE, used as responder cells, and finally co-cultured with effector cells. After 5 days incubation, cells were harvested and analyzed for flow cytometry by gating on the CFSE-labeled cells. The expression levels of Foxp3 and CD25 genes were detected by real-time polymerase chain reaction. Results After 9 days’ culture in vitro, the proportion of Foxp3+ γδ T cells was 7.87% in the blank group, which was significantly lower than that in the TGF-β, G-CSF and TGF-β + G-CSF group (62.3%, 52.9% and 63.5%) (P<0.001). The proportions of CD25+γδ T and CD27+γδ T cells were 3.9% and 3.5% in the blank group, which were also significantly lower than that in the TGF-β, G-CSF and TGF-β + G-CSF group (CD25+γδ T cells: 43.5%, 43.4% and 44.4%; CD27+γδ T cells: 41.5%, 41.9% and 44.0%, respectively) (P<0.001, P<0.001). The effector cells induced by anti-TCR γδ with TGF-β, G-CSF and TGF-β + G-CSF all manifested a more significant suppressive effect on responder cells than cells induced only by anti-TCR γδ. However, there was no significant difference in the effect of cell proliferation suppression in the TGF-β, G-CSF and TGF-β + G-CSF groups. The expression level of Foxp3 gene was 0.384% in the blank group, which was significantly lower than that in the TGF-β, G-CSF and TGF-β + G-CSF group (0.623%, 0.639% and 0.843%, respectively) (P<0.001). The expression level of Foxp3 in TGF-β+ G-CSF group was significantly higher than that in the TGF-β and G-CSF group (P=0.001, P=0.007). The expression level of CD25 gene was similar among the four groups (P=0.457). Conclusions The generation of γδ Tregs could be enhance in vitro induced by G-CSF. This may be one of the reasons that G-CSF play an immunoregulatory role on decreased GVHD onset during allo-HSCT. Disclosures: Wu: National Natural Science Foundation of China (No. 81200388); the Technology Plan of Guangdong Province of China (No. 2012B031800403); the project of the Zhujiang Science & Technology Star of Guangzhou city (No. 2013027).: Research Funding.
20
van der Heyde, Henri C., Joan M. Batchelder, Matyas Sandor, and William P. Weidanz. "Splenic γδ T Cells Regulated by CD4+ T Cells Are Required To Control Chronic Plasmodium chabaudi Malaria in the B-Cell-Deficient Mouse." Infection and Immunity 74, no. 5 (May 2006): 2717–25. http://dx.doi.org/10.1128/iai.74.5.2717-2725.2006.
Abstract:
ABSTRACT Little is known about the function and regulation of splenic γδ T cells during chronic Plasmodium chabaudi malaria. The splenic γδ T-cell population continues to expand, reaching levels equal to 4 times the number of splenocytes in an uninfected mouse. Splenic γδ T cells from JH −/− mice with chronic malaria expressed Vγ1+ or Vδ4+ in the same ratio as uninfected controls with Vγ1 cells dominating, but the Vγ2 ratio declined about twofold. γδ T cells from G8 mice specific for the TL antigen increased only 2-fold in number, compared with 10-fold in BALB/c controls, but G8 γδ T cells failed to express the B220 activation marker. Elimination of the parasite by drug treatment caused a slow depletion in the number of splenic γδ, CD4+, and CD8+ T cells. Following challenge, drug-cured JH −/− mice exhibited nearly identical parasitemia time courses as naïve controls. Depletion of either CD4+ T cells or γδ T cells from chronically infected JH −/− mice by monoclonal antibody treatment resulted in an immediate and significant (P < 0.05) exacerbation of parasitemia coupled with a marked decrease in splenic γδ T-cell numbers. The number of CD4+ T cells, in contrast, did not decrease in mice after anti-T-cell receptor γδ treatment. The results indicate that cell-mediated immunity against blood-stage malarial parasites during chronic malaria (i) requires the continued presence of blood-stage parasites to remain functional, (ii) is dependent upon both γδ T cells and CD4+ T cells, and (iii) lacks immunological memory.
21
Nwaneshiudu, Adaobi, Weon-ju Jung, Alexander Tsygankov, Marina Rayevskaya, Emilia Oleszak, Allen Myers, and Chris Platsoucas. "Gamma-delta TCR+ T-cells in systemic sclerosis (135.37)." Journal of Immunology 184, no. 1_Supplement (April 1, 2010): 135.37. http://dx.doi.org/10.4049/jimmunol.184.supp.135.37.
Abstract:
Abstract T-cell involvement may be central to the pathogenesis of systemic sclerosis (SSc). Like αβ TCR+ T-cells, γδ TCR+ T-cell diversity during T-cell development results in a repertoire of different T-cells each with a unique TCR. Therefore the probability of finding identical TCR on defined T-cell populations is negligible, except in the context of an antigen-driven T-cell response. To determine whether γδ TCR+ T-cells are clonally expanded in skin biopsies and/or peripheral blood of patients with SSc (n=7), γ-chain (VγI & Vγ9) and δ-chain (Vδ1 & Vδ2) TCR transcripts were amplified by gene-specific PCR, followed by cloning and sequencing. We report the presence of substantial proportions of identical VγI-chain transcripts (14.3%-37.2%; p<0.05), and Vγ9 transcripts (24%-83.3%; p<0.05) in skin biopsies and/or PBMC of patients with SSc. Likewise, there were multiple identical Vδ1-chain transcripts (25%-50%; p<0.05), and Vδ2-chain transcripts (18.1%-80%; p<0.05) in the samples analyzed. Extensive clonal expansions of γ- and δ-chain TCR transcripts were identified in skin biopsies and peripheral blood of patients with SSc, demonstrating the presence of oligoclonal populations of γδ TCR+ T-cells in these patients. These γδ TCR+ T-cells may have undergone activation and clonal expansion in vivo in response to as yet unidentified antigens. Future studies to identify the antigens recognized by these clonally expanded γδ TCRs will facilitate better understanding of SSc pathogenesis.
22
Kosub, David A., Ginger Lehrman, Jeffrey M. Milush, Dejiang Zhou, Elizabeth Chacko, Amanda Leone, Shari Gordon, et al. "Gamma/Delta T-Cell Functional Responses Differ after Pathogenic Human Immunodeficiency Virus and Nonpathogenic Simian Immunodeficiency Virus Infections." Journal of Virology 82, no. 3 (November 28, 2007): 1155–65. http://dx.doi.org/10.1128/jvi.01275-07.
Abstract:
ABSTRACT The objective of this study was to functionally assess gamma/delta (γδ) T cells following pathogenic human immunodeficiency virus (HIV) infection of humans and nonpathogenic simian immunodeficiency virus (SIV) infection of sooty mangabeys. γδ T cells were obtained from peripheral blood samples from patients and sooty mangabeys that exhibited either a CD4-healthy (>200 CD4+ T cells/μl blood) or CD4-low (<200 CD4 cells/μl blood) phenotype. Cytokine flow cytometry was utilized to assess production of Th1 cytokines tumor necrosis factor alpha and gamma interferon following ex vivo stimulation with either phorbol myristate acetate/ionomycin or the Vδ2 γδ T-cell receptor agonist isopentenyl pyrophosphate. Sooty mangabeys were observed to have higher percentages of γδ T cells in their peripheral blood than humans did. Following stimulation, γδ T cells from SIV-positive (SIV+) mangabeys maintained or increased their ability to express the Th1 cytokines regardless of CD4+ T-cell levels. In contrast, HIV-positive (HIV+) patients exhibited a decreased percentage of γδ T cells expressing Th1 cytokines following stimulation. This dysfunction is primarily within the Vδ2+ γδ T-cell subset which incurred both a decreased overall level in the blood and a reduced Th1 cytokine production. Patients treated with highly active antiretroviral therapy exhibited a partial restoration in their γδ T-cell Th1 cytokine response that was intermediate between the responses of the uninfected and HIV+ patients. The SIV+ sooty mangabey natural hosts, which do not proceed to clinical AIDS, provide evidence that γδ T-cell dysfunction occurs in HIV+ patients and may contribute to HIV disease progression.
23
Koh, Ki-Ryang, Hirohisa Nakamae, Kensuke Ohta, Hongzhang Li, Hideo Koh, Takahiko Nakane, Yasunobu Takeoka, et al. "Possible Involvement of γδ-T Cells in the Development of Chronic Graft-Versus-Host Disease after Allogeneic Hematopoietic Stem Cell Transplantation." Blood 110, no. 11 (November 16, 2007): 1971. http://dx.doi.org/10.1182/blood.v110.11.1971.1971.
Abstract:
Abstract Background: Chronic graft-versus-host disease (cGVHD) remains the major cause of late morbidity and non relapse mortality after allogeneic hematopoietic stem cell transplantation (allo-HSCT). However, the pathogenesis of cGVHD is poorly understood. This study was designed to explore the immunological mechanism of cGVHD with comprehensive analysis of peripheral T cell subsets (CD4, CD8, Th1, Th2, γδ-T and NKT), dendritic cell (DC) subsets and serum 17 different cytokines. Methods: 20 patients (pts) with complete donor cell engraftment over 100 days after allo-HSCT were enrolled, including 6 pts without cGVHD (control) and 14 pts with cGVHD (5 limited/ 9 extensive). All of the pts with extensive cGVHD were under systemic immunosuppressive therapy, and the pts with limited cGVHD or without cGVHD were free from immunosuppressant. T cell subsets including CD4 (CD3+CD4+CD8−), CD8 (CD3+CD4−CD8+), Th1 (CD4+CXCR3+CCR4−), Th2 (CD4+CXCR3−CCR4+), γδ-T (CD3+TCRVδ2+), and NKT (CD3+CD161+) and DC subsets (CD11c+DC and CD123+DC) were determined with a flow cytometer. Serum cytokines, including IL–1β, IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-10, IL-12, IL-13, IL-17, TNF-α, IFN-γ, G-CSF, GM-CSF, MIP-1β, and MCP-1) were analyzed by Bio-Plex Cytokine Assay. Differences among extensive cGVHD, limited cGVHD and control were evaluated by one-way ANOVA followed by Bonferroni method as a post hoc test. Results: A significant difference was found only in γδ-T cells, but not in any of the other cells, including CD4, CD8, Th1, Th2, NKT, and DC subsets among extensive cGVHD, limited cGVHD and control. There was a significant decrease in the mean absolute number of γδ-T cells in extensive cGVHD (4.0 /μl, n=9, p<0.001) and limited cGVHD (16.0 /μl, n=5, p=0.01), compared to that in control (56.7 /μl, n=5). This marked reduction of γδ-T cells in cGVHD was found also in the mean percentage of γδ-T cells in CD3 T cells (%-γδ-T) as follows: extensive cGVHD (0.53%, n=9, p<0.0001), limited cGVHD (1.00%, n=5, p<0.001) in contrast with control (4.74%, n=5). On the other hand, dramatical increase in IL-6 was observed in extensive cGVHD (18.4pg/ml, n=5), in contrast with limited cGVHD (2.5pg/ml, n=3, p=0.02) or control (0.8pg/ml, n=5, p=0.004), demonstrating IL-6 involvement in the severity of cGVHD. Moreover, IL-10 as a kind of Th2 cytokines was likely to increase in extensive cGVHD (3.8pg/ml, n=5, p=0.02), compared to that in control (0.2pg/ml, n=5), suggesting Th2 involvement in extensive cGVHD. The other cytokines including IL-4, TNF-α, IFN-γ did not show any significant changes between cGVHD and control. Interestingly, a positive correlation was observed between IL-6 and the reciprocal of the percentage of γδ-T cells in CD3 T cells (1/%-γδ-T) (n=12, r=0.79, p=0.002) with all samples put together, suggesting γδ-T cells as a negative regulator of IL-6. Discussion: γδ-T cells have been reported to control the innate immune system and participate in anti-tumor or anti-infection immunity in a major histocompatibility complex-unrestricted manner. Our data suggest the possible involvement of the reduction of γδ-T cells in the pathogenesis of cGVHD, and the usefulness of monitoring γδ-T cells in predicting the development of cGVHD after allo-HSCT.
24
Drobyski, William R., and David Majewski. "Donor γδ T Lymphocytes Promote Allogeneic Engraftment Across the Major Histocompatibility Barrier in Mice." Blood 89, no. 3 (February 1, 1997): 1100–1109. http://dx.doi.org/10.1182/blood.v89.3.1100.
Abstract:
Abstract T cells that express the αβ T-cell receptor are thought to be the T-cell population primarily responsible for facilitating alloengraftment. The role of γδ+ T cells that comprise only a minority of mature T cells in promoting allogeneic engraftment, however, has not been extensively studied. The purpose of this study was to determine whether γδ T cells were capable of facilitating alloengraftment in murine recipients of major histocompatibility complex-mismatched marrow grafts. We developed a model where engraftment of C57BL/6 × 129/F2 (H-2b) marrow in sublethally irradiated (800 cGy) recipients (AKR/J, H-2k) is dependent on the presence of mature donor T cells in the marrow graft. In this model, donor T-cell engraftment was significantly augmented by as few as 1 × 105 αβ T cells. The role of γδ T cells was then investigated using transgenic donors (C57BL/6 × 129 background) in which a portion of the T-cell receptor–β chain gene was deleted by gene targeting so that these mice lack αβ T cells. Addition of 10 × 106 naive γδ T cells to T-cell depleted marrow grafts was required to significantly increase alloengraftment, although donor T cells averaged <50% of total splenic T cells. To determine whether higher doses of γδ T cells would improve donor engraftment and eradicate residual host T cells, γδ T cells were ex vivo expanded with a γδ T-cell–specific monoclonal antibody and interleukin-2 and then transplanted into irradiated recipients. Transplantation of ≥ 160 × 106 activated γδ T cells was necessary to consistently and significantly augment donor cell chimerism and enhance hematopoietic reconstitution when compared to control mice, but host T cells persisted in these chimeras. Addition of 2.5 × 104 mature αβ T cells, which alone were incapable of facilitating engraftment, to T-cell depleted marrow grafts containing 160 × 106 activated γδ T cells resulted in long-term (<100 day) complete donor engraftment, indicating that limiting numbers of αβ T cells were required in the marrow graft for the eradication of residual host T cells. Using serial weight curves and B-cell reconstitution as end points, clinically significant graft-versus-host disease was not observed in these chimeras under these experimental conditions. These data show that, whereas less potent than αβ T cells, γδ T cells are able to promote engraftment and enhance hematopoietic reconstitution in allogeneic marrow transplant recipients.
25
Meraviglia, Serena, Carmela La Mendola, Valentina Orlando, Francesco Scarpa, Giuseppe Cicero, and Francesco Dieli. "Vγ9Vδ2 T cells as a promising innovative tool for immunotherapy of hematologic malignancies." Oncology Reviews 4, no. 4 (December 14, 2011): 211. http://dx.doi.org/10.4081/oncol.2010.211.
Abstract:
The potent anti-tumor activities of γδ T cells, their ability to produce pro-inflammatory cytokines, and their strong cytolytic activity have prompted the development of protocols in which γδ agonists or ex vivo-expanded γδ cells are administered to tumor patients. γδ T cells can be selectively activated by either synthetic phosphoantigens or by drugs that enhance their accumulation into stressed cells as aminobisphosphonates, thus offering new avenues for the development of γδ T cell-based immunotherapies. The recent development of small drugs selectively activating Vγ9Vδ2 T lymphocytes, which upregulate the endogenous phosphoantigens, has enabled the investigators to design the experimental approaches of cancer immunotherapies; several ongoing phase I and II clinical trials are focused on the role of the direct bioactivity of drugs and of adoptive cell therapies involving phosphoantigen- or aminobisphosphonate-activated Vγ9Vδ2 T lymphocytes in humans. In this review, we focus on the recent advances in the activation/expansion of γδ T cells in vitro and in vivo that may represent a promising target for the design of novel and highly innovative immunotherapy in patients with hematologic malignancies.<br />
26
Yao, Yi, Queping Liu, Carly Elizabeth Martin, Li Zhou, and Qing-Sheng Mi. "Embryonic fate mapping uncovers the critic role of miRNAs in skin-resident γδ T cell ontogeny." Journal of Immunology 198, no. 1_Supplement (May 1, 2017): 215.3. http://dx.doi.org/10.4049/jimmunol.198.supp.215.3.
Abstract:
Abstract Epidermis-resident γδ T cells (termed dendritic epidermal T cells, DETCs) are the first T cells developing in embryonic thymus. Given lack of a fate mapping tool, DETC ontogeny remains poorly understood. Recently, colony stimulating factor 1 receptor (CSF1R) Cre reporter was used to fate map tissue resident macrophages. To study the role of miR-155 in embryonic resident macrophages, we generated Csf1rCremiR-155GFP mice in which both miR-155 and GFP expressions are induced by Csf1rCre. As expected, the majority of skin macrophages highly expressed GFP. Surprisingly, the epidermal Vγ3+DETCs that emerged at E16.5 were all GFP+ cells, and nearly all fetal thymic γδ T cells were also GFP+, although overexpression of miR-155 did not disturb embryonic macrophages and γδ T cells. This observation was confirmed in Csf1rCreRosa26EGFP mice, suggesting CSF1R promoter driving both embryonic myeloid and lymphoid cell lineages. To further study the role of miRNAs in γδ T cell ontogeny, we next generated Csf1rCreDicerKO mice in which miRNAs are deficient. DETCs were almost completely eliminated in Dicer. KO epidermis at E18.5. The number of thymic Vγ3+ T cells were severely reduced (P < 0.0001) in KO embryos, and the frequency of annexin V+ apoptotic Vγ3+ T cells was raised (P < 0.001). Moreover, CD122+ or CD45RB+ mature Vγ3+ T cells were decreased (P < 0.01) in KO embryos, while immature CD24+Vγ3+ T cells were increased (P < 0.01). Thus, loss of miRNAs contributes to defects on survival and maturation of fetal thymic Vγ3+T cells, leading to DETC deficiency. Overall, this is the first study to demonstrate that Csf1rCre serves as a new tool for studying resident γδ T cell embryonic development and miRNAs regulate DETC ontogeny.
27
Tordesillas, Leticia, Junior Cianne, Jeremy S. Frieling, Xiomar Bustos, Conor C. Lynch, and Daniel Abate-Daga. "Abstract 1767: Biodistribution of zoledronate and effects on gd PSCA-CAR T cells in a model of bone metastatic prostate cancer." Cancer Research 83, no. 7_Supplement (April 4, 2023): 1767. http://dx.doi.org/10.1158/1538-7445.am2023-1767.
Abstract:
Abstract Background: Metastatic castrate resistant prostate cancer (mCRPC) is frequently manifested in the bone, leading to increased morbidity and mortality. We have previously demonstrated that γδ Chimeric Antigen Receptor (CAR) T cells targeting prostate stem cell antigen (PSCA) led to significant regression of established prostate cancer cells in the bone. Regression was further increased by combination with the bisphosphonate zoledronate (ZOL), routinely administered to mCRPC patients to prevent bone loss. To further optimize the use of γδ CAR T cells for mCRPC, we aimed to determine the kinetics of γδ CAR T cell in a mouse model of bone metastatic prostate cancer, either as single treatment or in combination with ZOL. In addition, we identified the sites of systemic ZOL accumulation in an immune competent model that could potentially induce off-target effects of γδ CAR T cells. Methods: Male NSG mice were injected with C4-2B prostate cancer cells expressing PSCA and luciferase in the left tibia, while the right tibia received PBS. ZOL (25 μg/kg) was injected every other day subcutaneously in half of the mice and was discontinued one day prior to administering T cells. When tumors were established, mice received γδ PSCA-CAR T cells, γδ untransduced (UT) T cells or were left untreated. Bone marrow from tumor-bearing tibia, tumor-naive tibia, femur, spleen and blood were recovered at multiple time points, and the number of γδ T cells was analyzed by flow cytometry. To determine systemic ZOL uptake, C57BL/6 mice were injected with PTE-82 prostate cell line in both tibias. One week after injection, mice received ZOL-AF647 every other day for 2 weeks. Bone marrow, liver, kidney, peritoneal lavage, skin-draining lymph nodes, Peyer’s patches and spleen were recovered and analyzed by flow cytometry. Results: γδ CAR T cells showed a rapid accumulation in the bone marrow recovered from tumor-bearing tibias, with 3 times more cells than those from mice treated with γδ UT cells (p=0.0002). The number of γδ T cells peaked at 5 days post infusion and were still detected after 21 days. Increased γδ CAR T cell accumulation was not observed in tumor-free bones or in spleen or blood, suggesting a preferential localization of γδ CAR T cells at tumor sites. Treatment with ZOL did not significantly affect the number or phenotype of γδ T cells accumulated in bone. Analysis of ZOL distribution showed significant uptake by macrophages, specially from liver and peritoneal lavage (p<0.0001), with an average of 30% and 70% of macrophages showing positive staining for ZOL-AF647, respectively (p<0.0001). Conclusion: γδ PSCA-CAR T cells accumulate and get activated at tumor sites with limited distribution outside the bone, while their kinetics is not affected by ZOL treatment in the NSG xenograft model. Additional studies will be necessary to determine the impact of ZOL uptake by myeloid cells on γδ CAR T cells migration. Citation Format: Leticia Tordesillas, Junior Cianne, Jeremy S. Frieling, Xiomar Bustos, Conor C. Lynch, Daniel Abate-Daga. Biodistribution of zoledronate and effects on gd PSCA-CAR T cells in a model of bone metastatic prostate cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 1767.
28
Ho, Andrew, Hunter C. Jonus, and Kelly C. Goldsmith. "Abstract 911: Butyrophilin 3A2 expression plays a critical role in phosphoantigen-mediated γδ T cell cytotoxicity of neuroblastoma cells." Cancer Research 83, no. 7_Supplement (April 4, 2023): 911. http://dx.doi.org/10.1158/1538-7445.am2023-911.
Abstract:
Abstract The clinical use of antibodies targeting the diaslganglioside GD2 improved survival and clearly identified a role for immunotherapy in the aggressive pediatric solid tumor neuroblastoma (NB). Even so, the 5-year survival rate for high risk patients remains 50%, warranting novel immunotherapeutic approaches. Given their MHC-independent cytotoxic activity towards transformed cells, γδ T cells are attractive tools for adoptive cellular immunotherapy. γδ T cell cytotoxicity is mediated through the expression of various activating receptors that contribute to multiple mechanisms of anti-tumor activity. Specifically, their expression of the γδ-TCR works in concert with tumor expression of members of the butyrophilin-3 (BTN3A) subfamily for cytotoxicity in response to phosphoantigens (pAg). In other cancers, stimulation of pAg production through zoledronate (ZOL) supplementation promotes γδ T cell recognition/killing. We have identified that γδ T cells are differentially cytotoxic towards NB cell lines, suggesting variations in expression of key players responsible for γδ T cell susceptibility. This work serves to understand the significance of BTN3A members in NB, and their impact on the susceptibility of NB to γδ-TCR mediated killing. It is hypothesized that the pAg-mediated response can be augmented by increasing intracellular pAg via use of ZOL or by manipulating BTN3A members. To test this, NB cells were pre-treated with either vehicle or ZOL for 24 hours and then co-incubated with expanded γδ T cells for 4 hours at various effector:target (E:T) ratios. Apoptotic death was then determined for NB cells using Annexin-V/7-AAD staining. Across 8 tested NB cell lines, Kelly, NB-1643, NGP, SK-N-AS, SH-SY5Y, NLF, and SMS-SAN cells were resistant to γδ T cells at a 10:1 E:T ratio when untreated with ZOL. IMR-5 demonstrated 12-18% lysis at a 5:1 E:T. When NB cells were pre-treated with ZOL, γδ T cell cytotoxicity was enhanced towards all models except SK-N-AS. We identified three NB cell lines with differential susceptibility to γδ T cell killing, SK-N-AS<NLF<IMR5. Using a high-risk NB gene expression dataset, expression of important BTN members was queried, revealing IMR5 to have BT3.2low expression, NLF to have BT3.2mod expression, and SK-N-AS to have BT3.2high expression. BT3.2 expression levels was confirmed via western blotting. It has been postulated that BT3.2 may be a decoy receptor that interacts with γδ T cells without intracellular signaling, given it lacks the intracellular pAg sensor. Due to its high expression in SK-N-AS, a lentiviral system was used to knockdown BT3.2. SK-N-A-S shBT3.2 models had 48-55% lysis at a 5:1 E:T when untreated and 65-72% lysis when pre-treated with ZOL. In summary, the anti-NB γδ T cell response can be augmented with ZOL pre-treatment. Additionally, the expression of BT3.2 may be responsible for differential sensitivity of NB cell lines to γδ T cells. Citation Format: Andrew Ho, Hunter C. Jonus, Kelly C. Goldsmith. Butyrophilin 3A2 expression plays a critical role in phosphoantigen-mediated γδ T cell cytotoxicity of neuroblastoma cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 911.
29
de Guibert, Sophie, Jean-Baptiste Thibert, Céline Bonnaventure, Patricia Ame-Thomas, Céline Pangault, Thierry Fest, Thierry Lamy, and Karin Tarte. "Quantitative and Functional Alterations of the Gamma Delta T-Cell Subset within Follicular Lymphoma Microenvironment." Blood 110, no. 11 (November 16, 2007): 2601. http://dx.doi.org/10.1182/blood.v110.11.2601.2601.
Abstract:
Abstract T cells carrying a γδ TCR account for less than 5% of CD3pos T cells in healthy individuals but are key effectors of innate immunity through the recognition of some unprocessed nonpeptide antigens of both self and foreign origin. Whereas the Vδ2 subpopulation represents more than 70% of peripheral blood γδ T cells, the Vδ1 subset is mainly located in the mucosal tissue. Increasing evidence suggest that γδ T cells have potent antitumor activity and are implicated in the defense against some haematological and epithelial malignancies. Moreover, Vδ2 T cells constitute an attractive immunotherapy strategy since they could be expanded and activated both in vivo and in vitro using synthetic phosphoantigens and aminobiphosphonates. Such strategies are currently tested in preliminary clinical trials, notably in follicular lymphoma (FL). However, an exhaustive phenotypic and functional characterisation of γδ T cells in this disease, including tumor infiltration, is still lacking. We first explored the composition of FL microenvironment using a multicolour flow cytometry analysis. We observed a significant decrease in the percentage of myeloid (LinnegCD11cposHLADRpos) and plasmacytoid (LinnegCD123posHLADRpos) dendritic cells (P = .0011 and P < .0001, respectively) in FL compared to normal secondary lymphoid organs. In addition, among CD3pos T cells, the proportion of follicular helper T cells (CD4posCXCR5posICOShi) was increased (P = .001) whereas regulatory T-cell (CD4posCD25posfoxp3pos) frequency was not altered. When considering the γδ T-cell compartment, we first highlighted a reduction of the Vδ2 subset in normal tonsils (Vδ2 = 23.48 ± 0.15% of γδ T cells, n = 11) when compared with peripheral blood. Remaining non-δ2 γδT cells were predominantly δ1 T cells. More importantly, infiltrating γδ T cells were significantly decreased in lymph node biopsies from FL patients (mean = 0.48 ± 0.4% of CD3pos T cells; n = 27) when compared both to normal tonsils (mean = 2.49 ± 1.6% of CD3pos T cells; n = 33) (P < .0001) and reactive lymph nodes (mean = 2.64 ± 2.6% of CD3pos T cells; n = 9) (P = .0009). This reduction affected both the Vδ1 and Vδ2 T-cell subsets. The functionality of γδ T cells was then assessed by the measurement of cell expansion and production of IFN-γ upon stimulation with the isopentenyl pyrophosphate (IPP) phosphoantigen. Amplification rate in vitro reached 14.6 ± 4.6 fold in tonsils (n = 10) but only 4.36 ± 1.9 fold in FL samples (n = 7) (P < .002) after 5 days of culture in the presence of IPP + IL-2 + IL-15. When focusing on the δ2 subset, this difference was further increased with a 40-fold amplification in tonsil and a 3-fold amplification in FL samples (P = .0004). Evaluation of IFN-γ production using ELISPOT assay revealed a high heterogeneity among tumor samples since 1 to 40% of δ2 T cells were able to respond to IPP stimulation (n = 7). Preliminary data argued for an association between the quantity and the functionality of γδ T cells in FL tumors. In conclusion, we reported an alteration of γδ T cell frequency and functionality within FL tumor niche. The next purpose will be to correlate these in vitro defects with in vivo clinical responses to immunotherapy strategies targeting γδ T cells.
30
Jimenez-Martinez, Maria C., Miguel Alonso, Iris Estrada-Garcia I, Sergio Estrada-Parra, Mayra Perez-Tapia, and Yonathan Garfias. "CXCL9 and γδ T cells are increased in patients with allergic conjunctivitis (P3159)." Journal of Immunology 190, no. 1_Supplement (May 1, 2013): 43.41. http://dx.doi.org/10.4049/jimmunol.190.supp.43.41.
Abstract:
Abstract Allergic conjunctivitis (AC) is a frequent disease that affects the ocular surface, The study of immune physiopathology of allergic conjunctivitis is focused on T helper cells. Recently it has been suggested that γδ T lymphocytes are necessary for the maintenance of the ocular allergic response, however there are not studies in humans about the role of γδ T lymphocytes in human AC. The aim of this study was to evaluate the frequency of γδ T cells and chemokines in tears of AC patients. Patients: 31 patients were included (16 men and 15 women), AC diagnosis was based on clinical history, physical and eye examination. PBMC were obtained from patients and healthy controls and frequency of γδ T cells was evaluated by flow cytometry; tear samples were also collected to determine chemokine concentrations by CBA. Results: γδ T lymphocytes were 11.9 times increased in AC-patients (22.63 ± 2.9%) compared to healthy donors (1.9 ± 1.9%) (p=0.002), chemokine determination in tears showed increased MIG/CXCL9 in AC patients (1604 ± 1404 pg/ml) when compared with healthy subjects (568 ±841pg/ml) p< 0.02. Conclusion: γδ T cells could be also involved in human allergic conjunctivitis as is has been demonstrated in animal models, it is possible that γδ T cells could migrate to conjunctiva due to several chemokines such as CXCL9, significant increased in AC patients, sustaining the local inflammation in conjunctiva
31
Weidanz, William P., GayeLyn LaFleur, Andrew Brown, James M. Burns, Irene Gramaglia, and Henri C. van der Heyde. "γδ T Cells but Not NK Cells Are Essential for Cell-Mediated Immunity against Plasmodium chabaudi Malaria." Infection and Immunity 78, no. 10 (July 26, 2010): 4331–40. http://dx.doi.org/10.1128/iai.00539-10.
Abstract:
ABSTRACT Blood-stage Plasmodium chabaudi infections are suppressed by antibody-mediated immunity and/or cell-mediated immunity (CMI). To determine the contributions of NK cells and γδ T cells to protective immunity, C57BL/6 (wild-type [WT]) mice and B-cell-deficient (JH−/− ) mice were infected with P. chabaudi and depleted of NK cells or γδ T cells with monoclonal antibody. The time courses of parasitemia in NK-cell-depleted WT mice and JH−/− mice were similar to those of control mice, indicating that deficiencies in NK cells, NKT cells, or CD8+ T cells had little effect on parasitemia. In contrast, high levels of noncuring parasitemia occurred in JH−/− mice depleted of γδ T cells. Depletion of γδ T cells during chronic parasitemia in B-cell-deficient JH−/− mice resulted in an immediate and marked exacerbation of parasitemia, suggesting that γδ T cells have a direct killing effect in vivo on blood-stage parasites. Cytokine analyses revealed that levels of interleukin-10, gamma interferon (IFN-γ), and macrophage chemoattractant protein 1 (MCP-1) in the sera of γδ T-cell-depleted mice were significantly (P < 0.05) decreased compared to hamster immunoglobulin-injected controls, but these cytokine levels were similar in NK-cell-depleted mice and their controls. The time courses of parasitemia in CCR2−/− and JH−/− × CCR2−/− mice and in their controls were nearly identical, indicating that MCP-1 is not required for the control of parasitemia. Collectively, these data indicate that the suppression of acute P. chabaudi infection by CMI is γδ T cell dependent, is independent of NK cells, and may be attributed to the deficient IFN-γ response seen early in γδ T-cell-depleted mice.
32
Yu, Xiaoqing, Song Li, Ling Cen, and Xuefeng Wang. "Abstract 3557: A pancancer gamma delta T cell repertoire atlas." Cancer Research 84, no. 6_Supplement (March 22, 2024): 3557. http://dx.doi.org/10.1158/1538-7445.am2024-3557.
Abstract:
Abstract While representing only a small portion of T cell population, Gamma delta (γδ) T cells have unique contributions to both innate and adaptive immunity. Compared to αβ T, γδ T cells have a more diverse TCR repertoire, allowing them to recognize a broader range of tumor antigens. Recent breakthrough discoveries indicate that the antitumor efficacy of γδ T cells can be potentiated or rescued through targeted therapeutic approaches. Despite their functional and translational significance, a comprehensive understanding of the γδ T cell repertoire within tumor tissues has yet to be fully achieved.In this work, we generated a pan-cancer γδ T cell repertoire Atlas through reanalyzing RNA-seq data from more than 11,000 tumors in TCGA utilizing TRUST4, an advanced computational method for reconstruction of TCR repertoires. This allowed us to offer the most accurate and comprehensive examination of the γδ TCR landscape spanning 33 cancer types, marking it the largest collection of γδ clones on human cancer to date. We processed 660 billion RNAseq reads, identified ~3 million CDR3 reads, applied a series of stringent filtering criteria to remove biologically implausible, ambiguous sequences, and out-of-frame TCRs, which resulted in 22,205 unique γδ TCR clones from 6,751 tumors. TCR gene enrichment scores were calculated to represent overall expression of γδ in each tumor. The γδ enrichment is highly variable between and within cancer types. As expected, tumors traditionally classified as immune-cold (UVM and GBM) show the lowest scores. Cancers intrinsically related to lymphatic system (LAML, THYM and DLBC), exhibit the highest scores. Beyond THYM, the top five solid-tumor cancers carrying the highest enrichment are PRAD, KIRC, TGCT, LUAD and KICH. We further profiled the clone counts, spectrum of clone sizes, CDR3 length, V-J pair usage, and clonality in 33 cancers. Overall, a dominant presence of large clone segment is not observed in most cancers. The most prevalent V-J pairs are TRGV10/9/2-TRGJ1 for γ, and TRDV1-TRDJ1 for δ chain. Cancers including LAML, KIRC, LUAD, SKCM, CESC, LUSC, STAD, display a relatively higher proportion of intermediate to larger clones. THYM exists the highest γδ diversity, with the lowest clonality and most diverse V-J pairs. In addition, we evaluated the prognostics value of γδ genes by multivariate Cox regression, adjusting for age, sex, stage, and tumor purity, and identified 38 γδ genes associated with patient survival with p<0.05 in 25 cancer types. Finally, within HNSC, we observed higher γδ gene enrichment (p=1.6e-9) and clone diversity (p=0.014) in HPV+ vs. HPV- tumors. The γδ gene enrichment is also found associated with patients’ survival (p=0.002) in HPV+, but not in HPV- HNSC. Similarly, COAD tumors with high Microsatellite Instability (MSI) shows higher γδ gene enrichment and clone diversity compared to MSI low COAD. Collectively, our findings serve as a foundational resource for γδ T cell research in oncology. . Citation Format: Xiaoqing Yu, Song Li, Ling Cen, Xuefeng Wang. A pancancer gamma delta T cell repertoire atlas [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 3557.
33
Jia, Luo, Sara Alonso, Guojun Wu, Yan Lam, Liping Zhao, and Karen L. Edelblum. "A transmissible γδ intraepithelial lymphocyte hyperproliferative phenotype is associated with the intestinal microbiota." Journal of Immunology 206, no. 1_Supplement (May 1, 2021): 17.07. http://dx.doi.org/10.4049/jimmunol.206.supp.17.07.
Abstract:
Abstract Intraepithelial lymphocytes (IEL) expressing the γδ T cell receptor provide a first line of defense at the intestinal barrier, yet how the intestinal microbiota influences this sentinel population remains unclear. Since commensal-induced tonic type I interferon (IFN) signaling promotes lamina propria lymphocyte homeostasis, we hypothesized that IFNα/β receptor (IFNAR) signaling may contribute to maintenance of the γδ IEL compartment. Morphometric analysis revealed a 2-fold increase in the number of GFP+ γδ IELs in IFNAR KO mice compared to WT housed in a standard barrier facility (SBF). γδ IEL proliferation was enhanced 50% in SBF IFNAR KO mice relative to SBF WT. This increase in γδ IEL number was not observed in IFNAR KO mice rederived into a cleaner, enhanced barrier facility (EBF) indicating that the γδ IEL hyperproliferative (γδHYP) phenotype occurs independently of IFNAR signaling. The transfer of dirty bedding from SBF IFNAR KO mice to EBF IFNAR KO breeding cages was sufficient to induce the γδHYP phenotype in both the breeders and their offspring, and antibiotic treatment prevented this transmission. SBF WT and IFNAR KO mice were crossed to generate F2 littermates, which all exhibited the γδHYP phenotype regardless of genotype. Evidence of horizontal and vertical transmission of the γδHYP phenotype led us to analyze the fecal microbiota. Greater microbial diversity was observed in mice with the γδHYP phenotype relative to those that did not (Shannon Index, p<0.01). Moreover, 6 amplicon sequence variants (ASV) were strongly associated with the γδHYP phenotype (P=1.075e-05). Thus, we have serendipitously discovered a specific microbiota that is both necessary and sufficient to promote the expansion of the γδ IEL compartment.
34
Perko, Ross W., Paul Thomas, and Mari Hashitate Dallas. "Elevated Gamma Delta T Cell Recovery Following Hematopoietic Stem Cell Transplantation Associated with Improved Long Term Overall Survival in Pediatric Patients with Acute Leukemia." Blood 120, no. 21 (November 16, 2012): 227. http://dx.doi.org/10.1182/blood.v120.21.227.227.
Abstract:
Abstract Abstract 227 Recent studies show that accelerated γΔ T cell reconstitution after hematopoietic stem cell transplantation (HSCT) is associated with improved overall survival (OS) though the mechanisms have not been elucidated. Here, we confirm that early γΔ T cell recovery after HSCT is an independent predictor for improving OS and event free survival (EFS). Moreover, patients with robust γΔ T cell recovery after HSCT appear to be protected from increased risk of serious life threatening infections, and acute gut and chronic graft versus host disease (GVHD). More importantly, we show that γΔ T cell recovery dynamics are independent from those of classical αβ T cells. We evaluated 102 consecutive pediatric patients with acute leukemia undergoing HSCT at St. Jude Children's Research Hospital from 1996–2011. The median age of the patients was 10.5 ± 5.9 yrs. (range, 0.6–25.2) and median follow up was 2.7±1.8 yrs. (range 0.2–6.0). There were 57% males, 43% females and 59% with ALL and 41% with AML. There were 14 patients with elevated γΔ T cells (≥1.75×105 cells/ml) and 88 with low/normal γΔ T cells (<1.75×105 cells/ml). There were no significant differences between the two groups with respect to age, sex, disease or donor source, p=0.7, 0.5, 1 and 0.07 respectively. Fours years after HSCT, OS was significantly higher for patients in the elevated group compared to the patients in the low/normal group, 93% and 60%, respectively, p=0.0173. Survival without relapse or graft failure (EFS) was significantly higher in the elevated group compared to the low/normal group, 85.7% and 58.0%, respectively. Since T cell reconstitution following HSCT is age dependent, we determined if γΔ T cell recovery correlated with age and/or CD3+ cells. Multivariate analysis showed no correlation between the number of CD3+ and γΔ T cells. In fact, 13 of 14 patients that recovered with increased number of γΔ T cells had normal or low numbers of CD3+ cells. Thus, γΔ T cell recovery is not a simple correlate of T cell reconstitution. Because γΔ T cells play a central role in maintaining intestinal epithelium integrity, we evaluated the incidence of gut GVHD. We found a significant lower rate of gut GVHD in the elevated group compared to the low/normal group, 0% and 17% respectively. Furthermore, the number of γΔ T cells in patients with cGVHD (2.3 x105 cells/ml) was significantly lower compared to patients without cGVHD (6.2 x105 cells/ml), p=0.01. This suggests that γΔ T cell may protect against gut and cGVHD. Since accumulating evidence suggests that γΔ T cells contribute to both innate and adaptive immune responses during infections, we evaluated the rate and types of infection between the two groups. We found a significant lower incidence of infection in the elevated group compared to the low/normal group, 21% and 54% respectively p=0.02. Furthermore, the elevated group had only viral infections while the low/normal group had viral, bacterial and fungal infections. Recent studies suggested that γΔ T cells could contribute to surveillance of CMV reactivation after HSCT through cooperation with anti-CMV IgG. Evaluation of CMV infections found no significant decrease in the incidence of CMV infection in the elevated group compared to the low/normal group, 14% and 2%. In summary, this is the first reported study of γΔ T cell recovery after HSCT in pediatric patients and adds new insights into the role γΔ T cells by evaluating the relationship of the most common complications such as relapse, GVHD and infections. Disclosures: No relevant conflicts of interest to declare.
35
Beck, Benjamin, Richard D. Lopez, G. Yancey Gillespie, Hyung Kim, Gretchen Cloud, Larisa Pereboeva, and Lawrence S. Lamb. "Peripheral Blood γδ T Cell Response to High-Grade Glioma: Implications for Localized Adoptive Immunotherapy." Blood 120, no. 21 (November 16, 2012): 4114. http://dx.doi.org/10.1182/blood.v120.21.4114.4114.
Abstract:
Abstract Abstract 4114 Glioblastoma multiforme (GBM) remains impervious to existing therapy. We have recently established that GBM is vulnerable to recognition and lysis by γδ T cells using in vitro cytotoxicity testing and in a treatment model based on human γδ T cell therapy of intracranial xenografted human glioma cell line tumors in immunodeficient mice. We have also shown that circulating γδ T cell counts and function are significantly reduced in GBM patients at the time of diagnosis. In this study, we sought to resolve the paradox between these findings with a working hypothesis that γδ T cells are depleted by activation-induced cell death following an unsuccessful immune response to GBM. We monitored peripheral and tumor immune responses over time in a C57BL/6 mouse model after intracranial placement of 1 × 105 syngeneic GL261 glioma cells, methylcellulose carrier alone, or no treatment (n=10/group). Tumor-bearing mice uniformly develop aggressive infiltrating gliomas with pseudopalisading morphology and necrotic centers that are uniformly fatal within 30 days. At 11 days following tumor placement, mice were asymptomatic. Brain sections revealed the presence of small infiltrating gliomas in 100% of mice. Peripheral blood expansion of γδ T cells was significantly increased compared to mice that received only the methylcellulose vehicle (p=0.0003). T cell proliferation was specific only to the γδ T cell compartment and did not involve other T cell subtypes, A significant proportion of γδ T cells expressed Annexin-V (20%-45%) in contrast to 2–5% in controls (p<0.0001) indicative of progression to apoptotic death. Mice that received intracranial injection of methylcellulose alone showed no changess relative to untreated mice. Although GL261 gliomas expressed γδ T cell-recognized NKG2D ligands H60, MULT-1 and RAE-1, peripheral γδ T cells did not infiltrate the intracranial tumors nor confer a survival advantage when compared to identical experiments conducted in tumor-bearing γδ T cell-deficient (C57BL/6δ−/−) mice. Placement of extra-cranial GL261 flank tumors resulted in accumulation of T cells around the periphery of the tumor but no significant T cell invasion into the tumor parenchyma. However, when mice were treated with adoptive transfer of 1.5 × 106ex vivo expanded/activated syngeneic γδ T cells by stereotactic intracranial injection on the same day as GL261 tumor placement, local tumor development was inhibited. Mice that were injected only with saline following tumor placement developed large tumors at the injection site. These data suggest that depletion of γδ T cells previously seen in GBM patients is likely due to activation-induced cell death. Although glioma-reactive γδ T cell effectors are stimulated in the early phase of tumor formation, they are unable to successfully invade and thereby impede tumor progression. Finally, these findings suggest that treatment strategies directed toward both reduction of tumor-derived immunosuppression and localized post-resection adoptive γδ T cell-based therapy may provide an approach to minimal residual GBM. Disclosures: No relevant conflicts of interest to declare.
36
Harada, Takeshi, Qu Cui, Shingen Nakamura, Hirokazu Miki, Asuka Oda, Ryota Amachi, Masami Iwasa, et al. "Robust Induction Of Th1-Like γδ T Cells With Anti-Myeloma Activity By Lenalidomide In Combination With HMB-PP As Well As Zoledronic Acid." Blood 122, no. 21 (November 15, 2013): 129. http://dx.doi.org/10.1182/blood.v122.21.129.129.
Abstract:
Abstract Multiple myeloma (MM) still remains incurable even with the implementation of novel therapeutic modalities, leading to the idea to develop various forms of immunotherapies. In this regard, γδ T cells bearing Vγ9Vδ2 TCR expanded from peripheral blood mononuclear cells (PBMCs) have attracted attention as potent effectors available in a novel immunotherapy against MM. Human Vγ9Vδ2 γδ T cells can be expanded ex vivo by aminobisphosphonates in combination with IL-2, and effectively target and impair MM cells. However, MM cells appear to protect themselves from external insults by immune cells in a unique bone marrow microenvironment created by the accumulation of mesenchymal stem cells/bone marrow stromal cells (BMSCs) with defective osteoblastic differentiation and acid-producing osteoclasts. To improve the therapeutic efficacy of γδ T cells, therefore, we need to develop a maneuver to effectively enhance the expansion and activity of γδ T cells while disrupting the MM cell-bone marrow interaction. Lenalidomide (Len), a novel immunomodulatory anti-MM agent, shows pivotal anti-MM activity by targeting immune cells as well as the interaction of MM cells and their surrounding cells in the bone marrow. The present study was undertaken to explore the efficacy of Len in combination with zoledronic acid (Zol) or a precursor of isopentenyl pyrophosphate (IPP) (E)-4 hydroxy-3-methyl-but-2-enyl pyrophosphate (HMB-PP), a microbial antigen for Vγ9Vδ2 TCR, on the induction and expansion of Th1-like γδ T cells with enhanced cytotoxic activity against MM cells in the skewed bone marrow microenvironment in MM. When combined with Zol (1μM), clinically relevant doses of Len (around 1 μM) substantially expanded γδ T cells from PBMCs to the levels similar to IL-2 (100 U/ml). Len was able to expand γδ T cells more robustly in combination with HMB-PP (1 μM) than Zol from PBMCs from the majority of normal donors. However, Len alone did not show any significant effects on γδ T cell expansion and activation, suggesting a costimulatory role of Len on Zol or HMB-PP-primed γδ T cells. The surface expression of LFA-1, and the cytotoxicity-associated molecules NKG2D, DNAX accessory molecule-1 (DNAM-1; CD226) and TRAIL were up-regulated in the expanded γδ T cells. Although functional diversity has been demonstrated in γδ T cells expanded by various stimuli, Len in combination with either Zol or HMB-PP enhanced intracellular IFN-γ along with the surface NKG2D but not Foxp3 in γδ T cells at higher levels than IL-2, suggesting robust induction of Th1-like γδ T cells by Len. Importantly, γδ T cells expanded with the combinatory treatments with Len and Zol or HMB-PP exerted potent cytotoxic activity against MM cells but not normal cells surrounding MM cells in bone marrow samples from patients with MM. Such treatments with Len was able to maintain the cytotoxic activity of the γδ T cells against MM cells in acidic conditions with lactic acid, and restored their anti-MM activity blunted in the presence of BMSCs. Interestingly, the expanded γδ T cells markedly suppressed the colony formation in semi-solid methylcellulose assays of RPMI8226 and KMS-11 cells [81±1 (mean ± SD) vs. 0±0 and 40±1 vs. 16±4 colonies/dish, respectively, p<0.01], and decreased in size their side populations, suggesting targeting a drug-resistant clonogenic MM cells. These results collectively demonstrate that Len and HMB-PP as well as Zol are an effective combination for ex vivo expansion of Th1-like γδ T cells with potent anti-MM activity, and suggest that Len in combination with Zol may maintain their in vivo anti-MM activity in the bone marrow where MM cells reside. The present results warrant further study on Len-based immunotherapy with γδ T cells. Disclosures: No relevant conflicts of interest to declare.
37
Minculescu, Lia, Hanne Vibeke Marquart, Lars Peter Ryder, Ida Schjødt, Lone Smidstrup Friis, Brian Thomas Kornblit, Søren Lykke Petersen, et al. "Improved Relapse-Free Survival and Overall Survival in Patients with High Immune Reconstitution of Gamma Delta T Cells 2 Months after Allogeneic Hematopoietic Stem Cell Transplantation." Blood 132, Supplement 1 (November 29, 2018): 3396. http://dx.doi.org/10.1182/blood-2018-99-111777.
Abstract:
Abstract Introduction: The role of T cell receptor (TCR) γδ cells in allogeneic hematopoietic stem cell transplantation (HSCT) is becoming of increasing interest1,2. In contrast to conventional alloreactive TCR αβ cells, TCR γδ cells are believed to have anti-tumor effects without causing graft-versus-host disease (GVHD). We conducted a single-center, prospective study to assess the impact of early TCR γδ cell immune reconstitution on overall survival, relapse and acute GVHD after HSCT. Methods: From October 2015 to March 2017, 108 consecutive patients transplanted for malignant diseases at the Bone Marrow Transplant Unit, Department of Hematology, Copenhagen University Hospital, Rigshospitalet, were included in the study, table 1. Fresh blood samples days 28, 56, 91, 180 and 360 after transplantation were analyzed for absolute concentrations of CD3-, CD4- and CD8 positive T cells together with a multi-color flow cytometry panel with staining for TCRαβ, TCRγδ, Vδ1, Vδ2, CD3, CD4, CD8, HLA-DR, CD196, CD45RO, CD45RA, CD16, CD56, CD314 (NKG2D) and CD337 (NKp30) for immune phenotyping. Results: After a median of 673 (386-913) days, 28 (26%) patients had died from relapse (n=14) and from transplant-related-mortality(n=14), respectively. A total of 24 (22%) patients experienced relapse during the observation time with median time to relapse of 177 (56-778) days. Acute GVHD grade 2-4 was diagnosed in 38 (35%) of patients. Patients were divided into two groups by dichotomization at the median value of TCR γδ cell concentrations for Kaplan-Meier analyses of overall survival (OS), relapse-free survival (RFS) and cumulative incidence analyses (Gray's test for competing risks) of relapse and acute GVHD. Patients with high concentrations of TCR γδ cells 56 days after transplantation had significantly higher OS and RFS compared with patients with low concentrations, p<0.001 and p=0.005, respectively, figure 1. In Cox regression analyses with TCR γδ cell concentrations included as (log2-transformed) continuous variables, increasing concentrations of TCR γδ cells were significantly associated with increased OS and RFS, 0.72 (95% CI 0.58-0.87), p=0.001 and 0.82 (95% CI 0.68-0.98), p=0.03, respectively. High concentrations of TCR γδ cells remained significantly associated with increased OS and RFS in Cox regression multivariate analysis adjusted for disease stage at transplantation and conditioning regimen, 0.66 (95% CI 0.53-0.683), p<0.001 (OS) and 0.79 (CI 95% 0.65-0.95), p=0.01 (RFS), respectively. In cumulative incidence analyses of death from relapse with death from transplant-related-mortality as a competing event, patients with high concentrations of TCR γδ cells 56 days after transplantation had a significantly lower risk of dying from relapse compared to patients with low concentrations, p=0.003, figure 2a. Cumulative incidence of acute GVHD with death as a competing event furthermore showed that patients with high concentrations of TCR γδ cells 28 days after transplantation had significantly less acute GVHD compared with patients with low concentrations, p=0.02, figure 2b. All associations between TCR γδ cell concentrations and outcomes were independent of the total CD3 T cell concentrations. Conclusion: The results of this prospective study suggest a protective effect of early robust TCR γδ cell immune reconstitution on relapse and acute GVHD resulting in increased OS after HSCT, and support further research in adoptive TCR γδ cell therapy in transplant patients. Handgretinger, R. & Schilbach, K. The potential role of gd T cells after allogeneic HCT for leukemia. Blood131, 1063-1072 (2018). Scheper, W., Grunder, C., Straetemans, T., Sebestyen, Z. & Kuball, J. Hunting for clinical translation with innate-like immune cells and their receptors. Leukemia28, 1181-1190 (2014). Disclosures No relevant conflicts of interest to declare.
38
Reyes, Ryan Michael, Yilun Deng, Deyi Zhang, Niannian Ji, Neelam Mukherjee, Karen Wheeler, Harshita B. Gupta, et al. "CD122-directed interleukin-2 treatment mechanisms in bladder cancer differ from αPD-L1 and include tissue-selective γδ T cell activation." Journal for ImmunoTherapy of Cancer 9, no. 4 (April 2021): e002051. http://dx.doi.org/10.1136/jitc-2020-002051.
Abstract:
BackgroundAnti-programmed death-ligand 1 (αPD-L1) immunotherapy is approved to treat bladder cancer (BC) but is effective in <30% of patients. Interleukin (IL)-2/αIL-2 complexes (IL-2c) that preferentially target IL-2 receptor β (CD122) augment CD8+ antitumor T cells known to improve αPD-L1 efficacy. We hypothesized that the tumor microenvironment, including local immune cells in primary versus metastatic BC, differentially affects immunotherapy responses and that IL-2c effects could differ from, and thus complement αPD-L1.MethodsWe studied mechanisms of IL-2c and αPD-L1 efficacy using PD-L1+ mouse BC cell lines MB49 and MBT-2 in orthotopic (bladder) and metastatic (lung) sites.ResultsIL-2c reduced orthotopic tumor burden and extended survival in MB49 and MBT-2 BC models, similar to αPD-L1. Using antibody-mediated cell depletions and genetically T cell-deficient mice, we unexpectedly found that CD8+ T cells were not necessary for IL-2c efficacy against tumors in bladder, whereas γδ T cells, not reported to contribute to αPD-L1 efficacy, were indispensable for IL-2c efficacy there. αPD-L1 responsiveness in bladder required conventional T cells as expected, but not γδ T cells, altogether defining distinct mechanisms for IL-2c and αPD-L1 efficacy. γδ T cells did not improve IL-2c treatment of subcutaneously challenged BC or orthotopic (peritoneal) ovarian cancer, consistent with tissue-specific and/or tumor-specific γδ T cell contributions to IL-2c efficacy. IL-2c significantly altered bladder intratumoral γδ T cell content, activation status, and specific γδ T cell subsets with antitumor or protumor effector functions. Neither IL-2c nor αPD-L1 alone treated lung metastatic MB49 or MBT-2 BC, but their combination improved survival in both models. Combination treatment efficacy in lungs required CD8+ T cells but not γδ T cells.ConclusionsMechanistic insights into differential IL-2c and αPD-L1 treatment and tissue-dependent effects could help develop rational combination treatment strategies to improve treatment efficacy in distinct cancers. These studies also provide insights into γδ T cell contributions to immunotherapy in bladder and engagement of adaptive immunity by IL-2c plus αPD-L1 to treat refractory lung metastases.
39
Robak, Ewa, Jerzy Z. Błoński, Jacek Bartkowiak, Hanna Niewiadomska, Anna Sysa-Jędrzejowska, and Tadeusz Robak. "Circulating TCR γδ Cells in the Patients with Systemic Lupus Erythematosus." Mediators of Inflammation 8, no. 6 (1999): 305–12. http://dx.doi.org/10.1080/09629359990315.
Abstract:
Systemic lupus erythematosus (SLE) is a disorder with a wide range of immunological abnormalities. The results of the studies undertaken in the last decade indicated that SLE pathogenesis was mainly connected with the breakdown of the activation control of B and T cells, generating humoral or cell-mediated responses against several self-antigens of affected cells. The last studies demonstrate that the role of γδ T lymphocytes in autoimmune diseases can be especially important. Flow cytometry techniques were used to investigate the number and percentage of TCR γδ T cells and their most frequent subtypes in peripheral blood of 32 patients with SLE and 16 healthy volunteers. We also correlated TCR γδ cells number with the level of T CD3+, T CD4+, T CD8+, and NK (CD16) cells (cytometric measurements) and SLE activity (on the basis of clinical investigations). Our studies were preliminary attempts to evaluate the role of that minor T cell subpopulation in SLE. Absolute numbers of cells expressing γδ TCR in most SLE blood specimens were significantly lower than in the control group (P<0.006). However, since the level of total T cell population was also decreased in the case of SLE, the mean values of the percentage γδ T cells of pan T lymphocytes were almost the same in both analysed populations (7.1% vs 6.3%, respectively). In contrast to Vδ2+ and Vγ9+ subtypes of pan γδ T cells, Vδ3+ T cells number was higher in SLE patients (20×10 cells/μl) than in healthy control group (2×2 cells/μl) (P=0.001). However, we found no differences between the numbers of pan γδ T lymphocytes and studied their subtypes in the patients with active and inactive disease. These cell subpopulations were doubled in the treated patients with immunosuppressive agents in comparison with untreated ones; however, data were not statistically significant. Our study indicated that Vδ3+ subtype of γδ T cells seems to be involved in SLE pathogenesis; however, we accept the idea that the autoimmunity does not develop from a single abnormality, but rather from a number of different events.
40
Dutt, Shelley, Anne Costanzo, Peggy Han, Kristen Taylor, Ken Fujioka, and Julie Jameson. "The impact of obesity on human γδ T cell function and homeostasis (HUM4P.271)." Journal of Immunology 194, no. 1_Supplement (May 1, 2015): 122.2. http://dx.doi.org/10.4049/jimmunol.194.supp.122.2.
Abstract:
Abstract CDC estimate the rate of obesity is on the rise, currently impacting one-third of the U.S. population. Obese patients exhibit side effects that include type 2 diabetes, chronic wounds and an increased vulnerability to infection. It is critical to understand how obesity impacts immune responses that affect these processes and create effective vaccines or antiviral treatments for obesity. We examined PBMC from donors with a lean BMI <25 or obese BMI >30. T cells were studied for responses to influenza virus A/PR/9/38 infection by monitoring upregulation activation markers and cytokine production, using flow cytometry. From these studies we have determined that obese donors have quantifiably fewer γδ T cells, which has implications for rapid immune responses to infection. γδ T cells are normally activated upon contact with infected influenza virus cells, however our findings show that IFN-γ production by γδ T cells is reduced in obese patients. In contrast there was no significant difference in the expression of cytotoxic markers such as granzyme B and NKG2D between lean and obese patients. Due to our finding, IL-2Ra is reduced on the cell surface and IL-2 is able to restore γδ T cell antiviral function,we hypothesize that γδ T cells in obese patients are lacking key T cell specific growth factor signals. Our findings show that gd homeostasis and antiviral function becomes impaired with obesity contributing to the increased susceptibility to infection found in obese subjects.
41
Liao, Winston W. P., K. S. Clifford Chao, Tom K. Hei, and Simon Cheng. "Association of IL17-expressing γδ t cells with acute radiation-induced pneumonitis." Journal of Clinical Oncology 30, no. 15_suppl (May 20, 2012): e21097-e21097. http://dx.doi.org/10.1200/jco.2012.30.15_suppl.e21097.
Abstract:
e21097 Background: Radiation pneumonitis is a substantial cause of morbidity and mortality with thoracic radiation for lung cancer. Little is known about the crucial mechanisms of the inflammatory response. We seek to determine if a key mediator of organ-specific inflammatory disorders and innate immune response, IL-17+ γδ T cells, is associated with radiation pneumonitis. Methods: C3HBe/FeJ mice (7 mice/group) were sham-irradiated as controls or exposed to a single dose of 15 Gy thoracic X-ray to develop pneumonitis. We have previously shown that TGFβ has an immunosuppressive activity in radiation pneumonitis. To potentiate the radiation pneumonitis, one group of mice was administered anti-TGFβ therapy with inhibitory TGFβ mAb (1D11, i.p.10 mg/kg/wk). Bronchoalveolar lavage fluid was assessed for cytology and inflammatory cytokine level. Lung tissues were examined for cell infiltration and histopathological changes. Cell surface marker and intracellular cytokine staining were performed on lymphocytes from the digested lungs by flow cytometry. Results: At 10 weeks post-irradiation, the lungs of the irradiated mice showed substantially more alveolar wall edema and increased infiltration of inflammatory cells compared with sham controls. Pneumonitis-involved lungs contained more IL-17+ γδ T cells (0.85% ± 0.00%) compared with sham controls (0.33% ± 0.02%), p<0.001. Furthermore increased IL-17+ γδ T cells were associated with potentiated radiation pneumonitis with anti-TGFβ therapy. There was a significant increased alveolar inflammation in irradiated mice injected with anti-TGFβ mAb. Anti-TGFβ irradiated lungs also contained significantly more IL-17+ γδ T cells (1.17% ± 0.13%) compared with irradiated controls (0.72% ± 0.13%), p<0.001. There was no increase of other TGFβ-dependent T cell subtypes such as IFNγ+ αβ T cells (Th1), IL-17+ αβ T cells (Th17), CD25+ Foxp3+ Tregs, nor activated macrophages in the potentiated pneumonitis lungs. Conclusions: Our findings implicate a novel role for IL17-expressing γδ T cells in radiation pneumonitis. This study reveals this innate immune response pathway as a potential target for therapeutic intervention in radiation lung injury
42
Roelandt, Philip R., Johan Maertens, Peter Vandenberghe, Chris Verslype, Tania Roskams, Raymond Aerts, Frederik Nevens, and Daan Dierickx. "Hepatosplenic γδ T-cell lymphoma after liver transplantation: Report of the first 2 cases and review of the literature." Liver Transplantation 15, no. 7 (June 26, 2009): 686–92. http://dx.doi.org/10.1002/lt.21748.
43
Cheung, Alice MS, Kar Wai Tan, Dian Yan Guo, Su-Ann Goh, Amanda SY Lau, Yeh Ching Linn, William YK Hwang, Yeow Tee Goh, and Shang Li. "In Vitro Expanded γδ T Cells Derived from Cord Blood Induce Potent and Specific Cytotoxicity Against Human Acute Myeloid Leukemia Cells." Blood 128, no. 22 (December 2, 2016): 2168. http://dx.doi.org/10.1182/blood.v128.22.2168.2168.
Abstract:
Abstract Conventional chemotherapeutic regimens for acute myeloid leukemia (AML) patients have demonstrated unsurpassed efficacy in the past decades, but are far from optimal with many patients experiencing multiple disease recurrence or intolerant of the intensive chemotoxicity. Variations in the treatment scheme as well as the use of alternative, targeted agents have been pursued with limited success. This is partly ascribed to the highly heterogeneous nature of the disease comprising a dynamic repertoire of evolving leukemic clones that are both molecularly and biologically diverse, making it difficult to achieve complete disease eradication without inducing adverse off-target effects. In this regard, cellular immunotherapy has emerged as a plausible alternative, leveraging on the diversity and degeneracy of the tumor antigen-recognizing receptor complex expressed by immune cells. In particular, there is a growing interest in the specific anti-leukemia efficacy of the innate-like γδ T cells, prompted by the association of an increased number of donor derived γδ T cells (specifically the Vδ1+ subtype) in allogeneic hematopoietic stem cell transplant (HSCT) patients with improved disease control in the absence of significant graft-versus-host disease (GvHD). We therefore hypothesize that these allogeneic γδ T cells exhibit potent leukemia specific cytotoxicity and serve as an effective treatment for AML. Given the rapid availability and widespread use of cord blood (CB) as an alternative for allogeneic HSCT, we first characterized and explored the potential of expanding CB-derived γδ T cells in vitro. Compared to mobilized peripheral blood (mPB), there is a significantly lower level of γδ T cell within CB mononuclear cells (MCs) (0.61% ± 0.36% in CB vs 4.95% ± 3.83% in mPB, p<0.001). However, the fraction of Vδ1+ subset within the γδ T cells in CB is >3.5-fold higher than that in mPB (56.05% ± 9.49% in CB vs 14.54% ± 12.2% in mPB, p<0.001). Importantly, while >90% of the Vδ1+ T cells in CB are of naive or central memory phenotype, more than 40% of these cells in mPB show effector memory expression. We established that optimal in vitro expansion of CB-derived γδ T cells requires direct contact to a mixture of irradiated PBMCs and Epstein-Barr virus-transformed lymphoblastoid cell line (EBV-LCL) at a fixed ratio in the gas-permeable G-Rex culture flask. Under these conditions, we were able to achieve up to 5,200-fold expansion of the starting γδ T cells over a period of 21 days. These cells exhibit potent in vitro cytotoxicity against a range of human AML cell lines, including K562, MOLM-14, MV4-11 and NOMO-1, as well as primary patient samples in a dose dependent manner. In contrast, there is minimal in vitro cytotoxicity against CD34+ cells isolated from allogeneic CB samples even at the highest effector-to-target cell (E:T) ratio tested. Infusion of the expanded γδ T cells into NOD/SCID/IL2Rγ-/- (NSG) mice at 3 weeks post-transplantation of a FLT3-ITD+ AML patient sample (P1) resulted in a significant decrease in leukemic cell engraftment in 40% of the γδ T cells-treated mice (87.46 ± 2.25% in non-treated vs 74.85 ± 1.55% in γδ T cells-treated mice, p=0.022). In a separate experiment, infusion into NSG mouse that was engrafted with low level (0.1%) of a different FLT3-ITD+ AML patient sample (P2) maintained the leukemic cell level low at 0.1% at 4 weeks post-infusion, as opposed to the >15-fold increase in leukemic burden (1.76%) seen in the untreated mouse. Consistent with our in vitro finding, infusion of up to 5 x 108 expanded CB derived γδ T cells/kg failed to induce severe GvHD symptoms in NSG mice engrafted with allogeneic human CB cells up to 8 weeks post-infusion, with no significant effect on the level of in vivo regenerated human myeloid and lymphoid cells, as well as colony-forming cells (CFCs). In summary, our data demonstrates that in vitro expanded CB derived γδ T cells show potent AML-specific cytotoxicity both in vitro and in vivo, making it a promising alternative cell source for immunotherapy. Further investigations to enhance the mechanistic understanding would be needed to seed for future clinical translation. Disclosures Hwang: Pfizer: Honoraria, Other: Travel support; MSD: Honoraria, Other: Travel support; BMS: Honoraria, Other: Travel support; Novartis: Honoraria, Other: Travel support; Celgene: Honoraria, Other: Travel support; Roche: Honoraria, Other: Travel support; Janssen: Honoraria, Other: Travel support; Sanofi: Honoraria, Other: Travel support.
44
Maggioli, Mayara Fernanda, Joyce Rodrigues Lobo, Maria Clorinda Soares Fioravanti, André Kipnis, and Ana Paula Junqueira-Kipnis. "Cellular immune response of Curraleiro Pé-duro and Nellore calves following Mycobacterium bovis-BCG vaccination." Pesquisa Veterinária Brasileira 33, no. 12 (December 2013): 1403–8. http://dx.doi.org/10.1590/s0100-736x2013001200002.
Abstract:
The present study aimed to assess the CD4, CD8 and γδ blood levels for Curraleiro Pé-duro, as well as the specific IFN-γ response after BCG vaccination using flow cytometry. The specific immune response against BCG was also evaluated by tuberculin skin test, performed before and 45 days after the vaccination. For comparison purposes, the same parameters were investigated on Nellore calves, an exotic bovine with resistance previously demonstrated. Naturally, Curraleiro Pé-duro animals had greater levels of CD4, CD8 and γδ lymphocytes (p<0.05). In response to vaccine, Curraleiro Pé-duro showed greater ability to respond specifically to BCG, generating resistance profile (Th1), evidenced by greater number of antigen specific CD4+ cells producing IFN-γ (p<0.05) and also higher tuberculin skin test reaction (p<0.05). Additionally, vaccinated Curraleiro Pé-duro calves had higher CD4 cells numbers than both Nellore control (p<0.05) and vaccinated groups (p<0.05). Curraleiro Pé-duro calves' higher basal lymphocytes blood level and stronger response in both IFN-γ and tuberculin skin test parameters probably play a positive role on protection/resistance to Mycobacterium bovis.
45
Toro, Jorge R., David J. Liewehr, Nina Pabby, Lynn Sorbara, Mark Raffeld, Seth M. Steinberg, and Elaine S. Jaffe. "Gamma-delta T-cell phenotype is associated with significantly decreased survival in cutaneous T-cell lymphoma." Blood 101, no. 9 (May 1, 2003): 3407–12. http://dx.doi.org/10.1182/blood-2002-05-1597.
Abstract:
The importance of αβ versus γδ T-cell subset antigen expression in the classification of peripheral T-cell lymphomas is still unclear. The objective of this study was to investigate the prognostic value of T-cell receptor–δ1 (TCRδ1) expression in primary cutaneous T-cell lymphomas. TCRδ1 cellular expression was assessed in skin biopsy specimens of 104 individuals with cutaneous T-cell lymphoma by immunohistochemistry. Both univariate (Kaplan-Meier) and multivariate (Cox regression) analyses were conducted to determine which variables (T-cell subtype, hemophagocytosis, histologic profile, age, sex, and adenopathy) were significantly associated with survival. Univariate analysis indicated that there was a statistically significant difference in survival between the patients with αβ cutaneous T-cell lymphoma and patients with γδ cutaneous T-cell lymphoma (P < .0001). There was also a statistically significant decrease in survival among patients who had subcutaneous involvement compared with patients who had epidermotropic and/or dermal involvement (P < .0001). Cox model analysis indicated that TCRδ1 expression was the factor that was most closely associated with decreased survival (P < .0001). Among those patients with cutaneous γδ T-cell lymphoma (n = 33), there was a trend for decreased survival for patients who had histologic evidence of subcutaneous fat involvement in comparison with patients who had epidermotropic or dermal patterns of infiltration (P = .067). No other prognostic factors were identified as having a notable association with outcome in this subgroup. TCRδ1 expression in primary cutaneous lymphomas is an independent prognostic factor associated with decreased survival.
46
Reyes, Ryan Michael, Yilun Deng, Deyi Zhang, Neelam Mukherjee, Niannian Ji, Karen Wheeler, Harshita B. Gupta, et al. "CD122-selective IL-2 complexes target γδ T and NK cells to reduce tumor-promoting Th17 effects and synergize with αPD-L1 to treat primary and metastatic bladder cancer." Journal of Immunology 204, no. 1_Supplement (May 1, 2020): 88.14. http://dx.doi.org/10.4049/jimmunol.204.supp.88.14.
Abstract:
Abstract αPD-L1 bladder cancer (BC) immunotherapy is effective in <30% of cases. To address the large αPD-L1-unresponsive subset of patients, we tested αIL-2/IL-2 complexes (IL-2c) that block IL-2 from binding high-affinity IL-2Rα (CD25) for preferential IL-2Rβ (CD122) binding. Regulatory T cells (Tregs) capture IL-2 by CD25 whereas CD8+T, γδ T, and NK cells use CD122. We hypothesized that the tumor microenvironment, including local immune cells in primary versus metastatic BC, differentially affects immunotherapy responses. We used PD-L1+ mouse BC cell lines MB49 and MBT-2, for intravesical ([IVe] in bladder) and intravenous (IV) challenge studies of local versus metastatic BC. αPD-L1 or IL-2c alone reduced tumor burden and extended survival in IVe MB49 and MBT-2. Treg depletion using FOXP3DTR mice further enhanced IVe IL-2c effects, consistent with the known tumor-promoting role of Tregs in human BC. Using in vivo cell depletion approaches, we found that γδ T cells and NK cells, but not CD8+ T cells, were necessary for IL-2c efficacy in bladder. γδ T cells also reduced intratumoral Th17 cells that promote MB49 growth and are elevated in human BC. We confirmed γδ T cell effects in δ TCR KO mice, which abrogated IL-2c efficacy but not αPD-L1 efficacy. Neither αPD-L1 nor IL-2c alone treated metastatic MB49 and MBT-2 BC but the combination improved survival in both. These data are consistent with our recent findings in human BC patients in whom γδ T cell and NK cell cytotoxicity improved BCG immunotherapy. Thus, IL-2c is a promising novel BC immunotherapy that can improve bladder-specific immunity in primary BC. In metastatic BC, combination with αPD-L1 may also be a successful BC treatment strategy due to engagement of innate and adaptive immune responses.
47
McKenna, Kyle C., Moriya Tsuji, Marcella Sarzotti, John B. Sacci, Adam A. Witney, and Abdu F. Azad. "γδ T Cells Are a Component of Early Immunity against Preerythrocytic Malaria Parasites." Infection and Immunity 68, no. 4 (April 1, 2000): 2224–30. http://dx.doi.org/10.1128/iai.68.4.2224-2230.2000.
Abstract:
ABSTRACT We tested the hypothesis that γδ T cells are a component of an early immune response directed against preerythrocytic malaria parasites that are required for the induction of an effector αβ T-cell immune response generated by irradiated-sporozoite (irr-spz) immunization. γδ T-cell-deficient (TCRδ−/−) mice on a C57BL/6 background were challenged with Plasmodium yoelii(17XNL strain) sporozoites, and then liver parasite burden was measured at 42 h postchallenge. Liver parasite burden was measured by quantification of parasite-specific 18S rRNA in total liver RNA by quantitative-competitive reverse transcription-PCR and by an automated 5′ exonuclease PCR. Sporozoite-challenged TCRδ−/− mice showed a significant (P < 0.01) increase in liver parasite burden compared to similarly challenged immunocompetent mice. In support of this result, TCRδ−/− mice were also found to be more susceptible than immunocompetent mice to a sporozoite challenge when blood-stage parasitemia was used as a readout. A greater percentage of TCRδ−/− mice than of immunocompetent mice progressed to a blood-stage infection when challenged with five or fewer sporozoites (odds ratio = 2.35, P = 0.06). TCRδ−/− mice receiving a single irr-spz immunization showed percent inhibition of liver parasites comparable to that of immunized immunocompetent mice following a sporozoite challenge. These data support the hypothesis that γδ T cells are a component of early immunity directed against malaria preerythrocytic parasites and suggest that γδ T cells are not required for the induction of an effector αβ T-cell immune response generated by irr-spz immunization.
48
Zorzeto, Tatiane Queiroz, Hisako Gondo Higashi, Marcos Tadeu Nolasco da Silva, Emilia de Faria Carniel, Waldely Oliveira Dias, Vanessa Domingues Ramalho, Taís Nitsch Mazzola, et al. "Immunogenicity of a Whole-Cell Pertussis Vaccine with Low Lipopolysaccharide Content in Infants." Clinical and Vaccine Immunology 16, no. 4 (March 4, 2009): 544–50. http://dx.doi.org/10.1128/cvi.00339-08.
Abstract:
ABSTRACT The lack of a clear correlation between the levels of antibody to pertussis antigens and protection against disease lends credence to the possibility that cell-mediated immunity provides primary protection against disease. This phase I comparative trial had the aim of comparing the in vitro cellular immune response and anti-pertussis toxin (anti-PT) immunoglobulin G (IgG) titers induced by a cellular pertussis vaccine with low lipopolysaccharide (LPS) content (wPlow vaccine) with those induced by the conventional whole-cell pertussis (wP) vaccine. A total of 234 infants were vaccinated at 2, 4, and 6 months with the conventional wP vaccine or the wPlow vaccine. Proliferation of CD3+ T cells was evaluated by flow cytometry after 6 days of peripheral blood mononuclear cell culture with stimulation with heat-killed Bordetella pertussis or phytohemagglutinin (PHA). CD3+, CD4+, CD8+, and T-cell receptor γδ-positive (γδ+) cells were identified in the gate of blast lymphocytes. Gamma interferon, tumor necrosis factor alpha, interleukin-4 (IL-4), and IL-10 levels in supernatants and serum anti-PT IgG levels were determined using enzyme-linked immunosorbent assay (ELISA). The net percentage of CD3+ blasts in cultures with B. pertussis in the group vaccinated with wP was higher than that in the group vaccinated with the wPlow vaccine (medians of 6.2% for the wP vaccine and 3.9% for the wPlow vaccine; P = 0.029). The frequencies of proliferating CD4+, CD8+, and γδ+ cells, cytokine concentrations in supernatants, and the geometric mean titers of anti-PT IgG were similar for the two vaccination groups. There was a significant difference between the T-cell subpopulations for B. pertussis and PHA cultures, with a higher percentage of γδ+ cells in the B. pertussis cultures (P < 0.001). The overall data did suggest that wP vaccination resulted in modestly better specific CD3+ cell proliferation, and γδ+ cell expansions were similar with the two vaccines.
49
Wan, Suigui, Chengcheng Zheng, Yang Lin, Hong Zhao, Li Su, and Changqing Xia. "<i>γδ</i>-T Large Granular Lymphocyte Leukemia Associated Hemaphagocytic Syndrome Complicated with Multiple Organ Dysfunction." Case Reports in Clinical Medicine 03, no. 04 (2014): 211–15. http://dx.doi.org/10.4236/crcm.2014.34049.
50
Li, Hua, Yubin Wang, and FuXiang Zhou. "Effect of ex vivo-expanded γδ-T cells combined with galectin-1 antibody on the growth of human cervical cancer xenografts in SCID mice." Clinical & Investigative Medicine 33, no. 5 (October 1, 2010): 280. http://dx.doi.org/10.25011/cim.v33i5.14353.
Abstract:
Objective: To investigate the antitumor activity of ex vivo-expanded γδ-T cells derived from tumor-infiltrating lymphocytes(γδTILs) from cervical cancer patients when combined with galectin-1 antibody and studied both in vitro and in vivo. Methods: The presence of γδTILs in cervical cancer specimens was detected by immunohistochemistry and γδTILs were expanded using the solid-phase antibody method. The expression of galectin-1 by the human cervical cancer cell line, SiHa, was measured by Western blot and ELISA. In vitro cytotoxic activities of expanded γδTILs, with or without galectin-1 inhibitor, were determined using the LDH-release test. In vivo antitumor activity of γδTILs, combined with galectin-1 antibody, was evaluated using the SCID mouse model. Results: γδTILs existed in the cervical cancer and the percentage of TCRγδ+ cells in γδTILs after ex vivo expansion was 91.2±1.2% detected by flow cytometry. SiHa cell expressed and secreted galectin-1 as measured by Western blot and ELISA. Expanded γδTILs from human cervical cancer demonstrated marked cytotoxicity to SiHa or Hela cells. In comparison with non-treated group, the cytotoxicity of γδ TILs towards SiHa or Hela cell was significantly increased when effector and target cells were incubated with either lactose or galectin-1 antibody at E/T ratio of 1:1 (p < 0.05). γδTILs, in combination with galectin-1 antibody treatment, significantly suppressed the growth of xenografts in SCID mice, in comparison with all other groups (p < 0.05). γδTILs alone also showed the ability to inhibit tumour growth in vivo, but were more efficient when combined with specific antibody (p < 0.05). Conclusion: Taken together, our results suggest that γδ-T cells, combined with galectin-1 antibody treatment, could be a more effective adoptive immunotherapy for patients with cervical cancer than traditional adoptive immunotherapy methods.