Journal articles on the topic 'ST2 antibodies'
To see the other types of publications on this topic, follow the link: ST2 antibodies.
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 'ST2 antibodies.'
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
Miller, Ashley M., Damo Xu, Darren L. Asquith, Laura Denby, Yubin Li, Naveed Sattar, Andrew H. Baker, Iain B. McInnes, and Foo Y. Liew. "IL-33 reduces the development of atherosclerosis." Journal of Experimental Medicine 205, no. 2 (February 11, 2008): 339–46. http://dx.doi.org/10.1084/jem.20071868.
Full textAbstract:
Atherosclerosis is a chronic inflammatory disease of the vasculature commonly leading to myocardial infarction and stroke. We show that IL-33, which is a novel IL-1–like cytokine that signals via ST2, can reduce atherosclerosis development in ApoE−/− mice on a high-fat diet. IL-33 and ST2 are present in the normal and atherosclerotic vasculature of mice and humans. Although control PBS-treated mice developed severe and inflamed atherosclerotic plaques in the aortic sinus, lesion development was profoundly reduced in IL-33–treated animals. IL-33 also markedly increased levels of IL-4, -5, and -13, but decreased levels of IFNγ in serum and lymph node cells. IL-33 treatment also elevated levels of total serum IgA, IgE, and IgG1, but decreased IgG2a, which is consistent with a Th1-to-Th2 switch. IL-33–treated mice also produced significantly elevated antioxidized low-density lipoprotein (ox-LDL) antibodies. Conversely, mice treated with soluble ST2, a decoy receptor that neutralizes IL-33, developed significantly larger atherosclerotic plaques in the aortic sinus of the ApoE−/− mice compared with control IgG-treated mice. Furthermore, coadministration of an anti–IL-5 mAb with IL-33 prevented the reduction in plaque size and reduced the amount of ox-LDL antibodies induced by IL-33. In conclusion, IL-33 may play a protective role in the development of atherosclerosis via the induction of IL-5 and ox-LDL antibodies.
2
Liu, Boyi, Yan Tai, Satyanarayana Achanta, Melanie M. Kaelberer, Ana I. Caceres, Xiaomei Shao, Jianqiao Fang, and Sven-Eric Jordt. "IL-33/ST2 signaling excites sensory neurons and mediates itch response in a mouse model of poison ivy contact allergy." Proceedings of the National Academy of Sciences 113, no. 47 (November 7, 2016): E7572—E7579. http://dx.doi.org/10.1073/pnas.1606608113.
Full textAbstract:
Poison ivy-induced allergic contact dermatitis (ACD) is the most common environmental allergic condition in the United States. Case numbers of poison ivy ACD are increasing due to growing biomass and geographical expansion of poison ivy and increasing content of the allergen, urushiol, likely attributable to rising atmospheric CO2. Severe and treatment-resistant itch is the major complaint of affected patients. However, because of limited clinical data and poorly characterized models, the pruritic mechanisms in poison ivy ACD remain unknown. Here, we aim to identify the mechanisms of itch in a mouse model of poison ivy ACD by transcriptomics, neuronal imaging, and behavioral analysis. Using transcriptome microarray analysis, we identified IL-33 as a key cytokine up-regulated in the inflamed skin of urushiol-challenged mice. We further found that the IL-33 receptor, ST2, is expressed in small to medium-sized dorsal root ganglion (DRG) neurons, including neurons that innervate the skin. IL-33 induces Ca2+ influx into a subset of DRG neurons through neuronal ST2. Neutralizing antibodies against IL-33 or ST2 reduced scratching behavior and skin inflammation in urushiol-challenged mice. Injection of IL-33 into urushiol-challenged skin rapidly exacerbated itch-related scratching via ST2, in a histamine-independent manner. Targeted silencing of neuronal ST2 expression by intrathecal ST2 siRNA delivery significantly attenuated pruritic responses caused by urushiol-induced ACD. These results indicate that IL-33/ST2 signaling is functionally present in primary sensory neurons and contributes to pruritus in poison ivy ACD. Blocking IL-33/ST2 signaling may represent a therapeutic approach to ameliorate itch and skin inflammation related to poison ivy ACD.
3
YOSHIDA, KAZUKO, TAKAO ARAI, TAKASHI YOKOTA, NORIO KOMATSU, YASUSADA MIURA, KEN YANAGISAWA, TSUNAO TETSUKA, and SHIN-ICHI TOMINAGA. "Studies on Natural ST2 Gene Products in the Human Leukemic Cell Line UT-7 Using Monoclonal Antihuman ST2 Antibodies." Hybridoma 14, no. 5 (October 1995): 419–27. http://dx.doi.org/10.1089/hyb.1995.14.419.
Full text
4
Chen, Mengjiao, Peijun Ding, Lili Yang, Xufeng He, Chunjie Gao, Guoxun Yang, and Huimin Zhang. "Evaluation of Anti-Inflammatory Activities of Qingre-Qushi Recipe (QRQS) against Atopic Dermatitis: Potential Mechanism of Inhibition of IL-33/ST2 Signal Transduction." Evidence-Based Complementary and Alternative Medicine 2017 (2017): 1–11. http://dx.doi.org/10.1155/2017/2489842.
Full textAbstract:
To evaluate the anti-inflammatory activities of QRQS against AD and the inhibitory molecular mechanisms of IL-33/ST2 signal transduction, BALB/c mice were divided into six groups (normal control, OVA control, low-dose of QRQS, middle-dose of QRQS, high-dose of QRQS, and cetirizine) and epicutaneously exposed to ovalbumin or PBS for 3 weeks and treated with QRQS for 2 weeks. Skin biopsies and blood samples were obtained for histological study, antibody analysis, and RNA isolation. HaCaT cells, stimulated by TNF-α and IFN-γ, were treated with QRQS to evaluate mRNA and protein expression by RT-PCR and ELISA. QRQS decreased both epidermal and dermal thickness, alleviated dermatitis, and reduced IL-33 and ST2 positive cell numbers. The concentration of specific IgE, IgG, IgG1, and IgG2a antibodies in serum and the expression of IL-33, ST2, IL-1RAcP, IL-4, and IL-13 mRNA in the skin were suppressed. No significant difference exists in TNF-α or IFN-γ. QRQS decreased IL-33 mRNA and protein secretion in HaCaT cells exposed to TNF-α and IFN-γ in a time- and concentration-dependent manner. QRQS regulates related molecule expression of ovalbumin-induced dermatitis involved in the IL-33/ST2 signaling axis in the treatment of acute AD.
5
Martin, Rebecca, Joseph Cornelius Lownik, Jared Farrar, Grayson Way, Matthew Zellner, Bin Ni, Francesco Celi, and Daniel H. Conrad. "Loss of ADAM17 from macrophages induces ST2+ T regulatory cells that limit obesity-induced metabolic inflammation in mice through changes in both membrane and soluble TNF." Journal of Immunology 208, no. 1_Supplement (May 1, 2022): 46.14. http://dx.doi.org/10.4049/jimmunol.208.supp.46.14.
Full textAbstract:
Abstract Obesity is known to contribute to changes in immune cells, cytokines and metabolism. Secreted TNF (sTNF) levels are directly correlated in obesity with weight gain and morbidity, as well as metabolic dysregulation. ADAM17 cleaves transmembrane TNF (mTNF) into its soluble form. This is well defined, but the mechanism behind the regulation of TNF, the source of TNF, as well as the downstream action of TNF is less clear. Our data showed that mice with ADAM17 deleted from the myeloid lineage (ADAM17LysM) had a significant metabolic advantage as illustrated by an increase in glucose tolerance and decrease in body weight. Immunophenotyping of the fatpad of these mice using high dimensional flow cytometry after high fat diet (HFD) feeding showed an increase in activated ST2+ T regulatory cells (Tregs). Based on these findings, we hypothesized that modulation of TNF on macrophages through loss of ADAM17, helps maintain a healthy metabolic state through recruitment and activation of ST2+ Treg cells that limit metabolic dysregulation. Next, we showed that the accumulation and activation of these Tregs occurred through two separate mechanistic actions of TNF. First, the loss of sTNF production from macrophages decreased soluble ST2 (sST2) production by adipocytes. This loss of sST2 augments ST2+ Treg recruitment and expansion in the fatpad due to increased bioavailable IL-33. These experiments were controlled using a myeloid-specific knockout of TNF. Second, utilizing TNFR blocking antibodies, we demonstrated that ST2+ Treg activation was dependent on macrophage mTNF acting through TNFR2. Supported by grants from NIH (R56 AI139658 and R01 AI018697)
6
Fu, Denggang, Hua Jiang, Alan Long, Hong fen Guo, Maegan L. Capitano, Baskar Ramdas, Reuben Kapur, Nai-Kong V. Cheung, and Sophie Paczesny. "Immunotherapy Targeting ST2/IL-33 Signaling in Myeloid Leukemia Stem Cells." Blood 138, Supplement 1 (November 5, 2021): 23. http://dx.doi.org/10.1182/blood-2021-149672.
Full textAbstract:
Abstract Therapies for acute myeloid leukemia (AML) has barely changed over 30 years, while treatment for other blood cancers have made remarkable leaps forward. Recent advances in genomics have allowed molecular targeted therapies (i.e. FLT3-ITD, IDH, c-KIT inhibitors) extending survival, but most patients still succumb (Burd et al. Nat Med 2020). Therefore, developing more efficient, less toxic, and immune-based therapies for AML is an urgent unmet need. Previous studies showed that stromal cell-derived IL-33 stimulates myeloproliferative neoplasms (Mager et al. J Clin Invest 2015). Stimulation-2 (ST2), IL-33 receptor, contributes to leukemia stem cells (LSCs) survival in Cbfb-MYH11 mice (Wang et al . Sci Rep 2019). We showed that ST2 blockade enhanced graft versus leukemia activity against MLL-AF9 egfp AML after hematopoietic cell transplantation (Zhang et al . Sci Transl Med 2015). We and others, also, found that ST2 is expressed on normal murine and human hematopoietic stem cells (HSCs), respectively (Capitano et al. Blood Cells Mol Dis 2020; Alt et al. Biol Blood Marrow Transplant 2019). These data suggest a leukemia-promoting role of ST2/IL-33 signaling. To determine clinical relevance of ST2 in AML, we generated Kaplan-Meier curves using TCGA (n=173) and TARGET AML (n=187) databases. Decreased survival was observed in patients with high IL1RL1 (ST2 gene) which was validated in an independent database (AMLCG 1999 trial, n=417) (Fig. 1A). Since ST2 is expressed on HSCs, we interrogated if ST2 is expressed on LSCs defined as CD34 +CD38 - in the Princess Margaret Leukemia biobank (n=192), and found ST2 is higher on LSCs vs CD34 -CD38 +/- cells (Fig. 1B). We then sought to analyze ST2 on bone marrow samples comparing complete responders vs refractory patients to note that ST2 expression was increased in refractory patients' LSCs (Fig. 1C). To scrutinize the role of ST2 in initiating leukemogenesis, we performed limiting dilution transplantation using 500, 200, and 50 Lin -Sca-1 +c-KIT +-sorted LSCs from WT vs ST2 -/- MLL-AF9 egfp transduced cells. Frequency of LSCs in ST2 -/- cells was decreased by ~15-fold as opposed to WT cells [1:2141 (1:546-1:8405) vs 1:145 (1:75-1:283), p=3.37e-05] (Fig. 2A). We also tested leukemia maintenance, secondary transplantations from the primary recipients resulted in leukemia growth delay in ST2 -/- vs WT cells which was confirmed in tertiary transplantations (Fig. 2B-E). Self-renewal ability of LSCs is correlated to reactive oxygen species (ROS) (Testa et al. Exp Hematol 2016), and we found that ROS levels in ST2 -/- leukemic cells are markedly diminished in contrast to WT leukemic cells (Fig. 2F). ST2 deficiency in leukemic cells arrests G2/S/M cell cycle progression in LSCs (Fig. 2G-J). These data indicated that ST2 is indispensable for initiating and maintaining LSCs in MLL-AF9 AML. We next developed murine and human Fc-silenced-bispecific antibodies engaging mouse or human ST2 and CD3 (BsAb) built on the IgG[L]-scFv platform with proven ability to drive T cells into human tumors for effective tumor ablation (Santich et al. Sci Transl Med 2020; Park et al. J Immunother Cancer 2021) (Fig. 3A, 3E). Both BsAbs showed >90% purity by HPLC, stability under heat stress and low endotoxin. Animals did not exhibit any in vivo toxicity at BsAb doses of 0.4, 2, 5, 10 μg ip q 3 days x 6 doses (not shown). In the immunocompetent MLL-AF9 mice, murine anti-ST2 BsAb (BC281) treatment (10 μg i.p, 4 days post-AML challenge and given every three days for a total of 6 injections) resulted in extended survival compared to isotype control mice (Fig. 3B). Leukemic cells and LSCs were accordingly decreased in treated vs control group (Fig. 3C-D). We modeled humanized leukemic mice with MOLM-14 egfp cells and weekly injection of human CD8 + T cells in NOD.Cg-Prkdc scid Il2rg tm1Wjl/SzJ (NSG) mice (Fig. 3F). Animals treated with human anti-ST2 BsAb (BC282), using a similar regimen as for the immunocompetent model, led to better survival when compared to animals treated with mutated non-functional anti-ST2 BsAb (BC283) (Fig. 3G). Frequency of MOLM-14 egfp cells was lower in the BC282 vs BC283 group (Fig. 3H). These results suggested that anti-ST2 BsAbs can inhibit AML growth to improve survival. We concluded that ST2 is a potential therapeutic target, and ST2-specific T cell engaging BsAbs represent promising immunotherapeutics for AML. Figure 1 Figure 1. Disclosures Cheung: Medical University of South Carolina: Patents & Royalties: inventor on the ST2 bispecific antibody patent application; Y-mabs Therapeutics and Abpro-Labs Inc: Patents & Royalties: inventor on multiple patents filed by MSK, including those licensed to Ymabs Therapeutics, Biotec Pharmacon, and Abpro-labs; Eureka Therapeutics: Membership on an entity's Board of Directors or advisory committees. Paczesny: Medical University of South Carolina: Patents & Royalties: inventor on the ST2 bispecific antibody patent application.
7
Huang, Qiong, Chen Duo Li, Yi Ran Yang, Xiao Feng Qin, Jing Jing Wang, Xin Zhang, Xiao Nan Du, et al. "Role of the IL-33/ST2 axis in cigarette smoke-induced airways remodelling in chronic obstructive pulmonary disease." Thorax 76, no. 8 (February 15, 2021): 750–62. http://dx.doi.org/10.1136/thoraxjnl-2020-214712.
Full textAbstract:
BackgroundEfficient therapy and potential prophylaxis are confounded by current ignorance of the pathogenesis of airway remodelling and blockade in COPD.ObjectiveTo explore the role of the IL-33/ST2 axis in cigarette smoke (CS) exposure-induced airways remodelling.MethodsC57BL/6, BALB/c and IL-1RL1-/- mice exposed to CS were used to establish an animal surrogate of COPD (air-exposed=5~8, CS-exposed=6~12). Hallmarks of remodelling were measured in mice. Cigarette smoke extract (CSE)-induced proliferation and protein production in vitro by fibroblasts in the presence of anti-interleukin-33 (anti-IL-33) or hST2 antibodies were measured. Expression of IL-33 and ST2 and other remodelling hallmarks were measured, respectively, in bronchoalveolar lavage fluid (BALF) (controls=20, COPD=20), serum (controls=59, COPD=90) and lung tissue sections (controls=11, COPD=7) from patients with COPD and controls.ResultsWild-type mice exposed to CS elevated expression of hallmarks of tissue remodelling in the lungs and also in the heart, spleen and kidneys, which were significantly abrogated in the IL-1RL1-/- mice. Fibroblasts exposed to CSE, compared with control, exhibited early cellular translocation of IL-33, accompanied by proliferation and elevated protein synthesis, all inhabitable by blockade of IL-33/ST2 signalling. Expression of IL-33 and ST2 and hallmarks of tissue remodelling were significantly and proportionally elevated in BALF, serum and tissue samples from patients with COPD.ConclusionsExposure to CS induces remodelling changes in multiple organs. The data support the hypothesis that CS-induced lung collagen deposition is at least partly a result of CS-induced IL-33 translocation and release from local fibroblasts.
8
Mohd Jaya, Fatin Nurizzati, Zhongyi Liu, and Godfrey Chi-Fung Chan. "Early Treatment of Interleukin-33 can Attenuate Lupus Development in Young NZB/W F1 Mice." Cells 9, no. 11 (November 10, 2020): 2448. http://dx.doi.org/10.3390/cells9112448.
Full textAbstract:
Interleukin-33 (IL-33), a member of the IL-1 cytokine family, has been recently associated with the development of autoimmune diseases, including systemic lupus erythematosus (SLE). IL-33 is an alarmin and a pleiotropic cytokine that affects various types of immune cells via binding to its receptor, ST2. In this study, we determine the impact of intraperitoneal IL-33 treatments in young lupus, NZB/W F1 mice. Mice were treated from the age of 6 to 11 weeks. We then assessed the proteinuria level, renal damage, survival rate, and anti-dsDNA antibodies. The induction of regulatory B (Breg) cells, changes in the level of autoantibodies, and gene expression were also examined. In comparison to the control group, young NZB/W F1 mice administered with IL-33 had a better survival rate as well as reduced proteinuria level and lupus nephritis. IL-33 treatments significantly increased the level of IgM anti-dsDNA antibodies, IL-10 expressing Breg cells, and alternatively-induced M2 macrophage gene signatures. These results imply that IL-33 exhibits a regulatory role during lupus onset via the expansion of protective IgM anti-dsDNA as well as regulatory cells such as Breg cells and M2 macrophages.
9
Michigami, Toshimi, Nobuaki Shimizu, Paul J. Williams, Maria Niewolna, Sarah L. Dallas, Gregory R. Mundy, and Toshiyuki Yoneda. "Cell–cell contact between marrow stromal cells and myeloma cells via VCAM-1 and α4β1-integrin enhances production of osteoclast-stimulating activity." Blood 96, no. 5 (September 1, 2000): 1953–60. http://dx.doi.org/10.1182/blood.v96.5.1953.
Full textAbstract:
Abstract Myeloma is a unique hematologic malignancy that exclusively homes in the bone marrow and induces massive osteoclastic bone destruction presumably by producing cytokines that promote the differentiation of the hematopoietic progenitors to osteoclasts (osteoclastogenesis). It is recognized that neighboring bone marrow stromal cells influence the expression of the malignant phenotype in myeloma cells. This study examined the role of the interactions between myeloma cells and neighboring stromal cells in the production of osteoclastogenic factors to elucidate the mechanism underlying extensive osteoclastic bone destruction. A murine myeloma cell line 5TGM1, which causes severe osteolysis, expresses α4β1-integrin and tightly adheres to the mouse marrow stromal cell line ST2, which expresses the vascular cell adhesion molecule-1 (VCAM-1), a ligand for α4β1-integrin. Co-cultures of 5TGM1 with primary bone marrow cells generated tartrate-resistant acid phosphatase-positive multinucleated bone-resorbing osteoclasts. Co-cultures of 5TGM1 with ST2 showed increased production of bone-resorbing activity and neutralizing antibodies against VCAM-1 or α4β1-integrin inhibited this. The 5TGM1 cells contacting recombinant VCAM-1 produced increased osteoclastogenic and bone-resorbing activity. The activity was not blocked by the neutralizing antibody to known osteoclastogenic cytokines including interleukin (IL)-1, IL-6, tumor necrosis factor, or parathyroid hormone-related peptide. These data suggest that myeloma cells are responsible for producing osteoclastogenic activity and that establishment of direct contact with marrow stromal cells via α4β1-integrin/VCAM-1 increases the production of this activity by myeloma cells. They also suggest that the presence of stromal cells may provide a microenvironment that allows exclusive colonization of myeloma cells in the bone marrow.
10
Michigami, Toshimi, Nobuaki Shimizu, Paul J. Williams, Maria Niewolna, Sarah L. Dallas, Gregory R. Mundy, and Toshiyuki Yoneda. "Cell–cell contact between marrow stromal cells and myeloma cells via VCAM-1 and α4β1-integrin enhances production of osteoclast-stimulating activity." Blood 96, no. 5 (September 1, 2000): 1953–60. http://dx.doi.org/10.1182/blood.v96.5.1953.h8001953_1953_1960.
Full textAbstract:
Myeloma is a unique hematologic malignancy that exclusively homes in the bone marrow and induces massive osteoclastic bone destruction presumably by producing cytokines that promote the differentiation of the hematopoietic progenitors to osteoclasts (osteoclastogenesis). It is recognized that neighboring bone marrow stromal cells influence the expression of the malignant phenotype in myeloma cells. This study examined the role of the interactions between myeloma cells and neighboring stromal cells in the production of osteoclastogenic factors to elucidate the mechanism underlying extensive osteoclastic bone destruction. A murine myeloma cell line 5TGM1, which causes severe osteolysis, expresses α4β1-integrin and tightly adheres to the mouse marrow stromal cell line ST2, which expresses the vascular cell adhesion molecule-1 (VCAM-1), a ligand for α4β1-integrin. Co-cultures of 5TGM1 with primary bone marrow cells generated tartrate-resistant acid phosphatase-positive multinucleated bone-resorbing osteoclasts. Co-cultures of 5TGM1 with ST2 showed increased production of bone-resorbing activity and neutralizing antibodies against VCAM-1 or α4β1-integrin inhibited this. The 5TGM1 cells contacting recombinant VCAM-1 produced increased osteoclastogenic and bone-resorbing activity. The activity was not blocked by the neutralizing antibody to known osteoclastogenic cytokines including interleukin (IL)-1, IL-6, tumor necrosis factor, or parathyroid hormone-related peptide. These data suggest that myeloma cells are responsible for producing osteoclastogenic activity and that establishment of direct contact with marrow stromal cells via α4β1-integrin/VCAM-1 increases the production of this activity by myeloma cells. They also suggest that the presence of stromal cells may provide a microenvironment that allows exclusive colonization of myeloma cells in the bone marrow.
11
Burt, Philipp, Rebecca Cornelis, Gustav Geißler, Stefanie Hahne, Andreas Radbruch, Hyun-Dong Chang, and Kevin Thurley. "Data-Driven Mathematical Model of Apoptosis Regulation in Memory Plasma Cells." Cells 11, no. 9 (May 5, 2022): 1547. http://dx.doi.org/10.3390/cells11091547.
Full textAbstract:
Memory plasma cells constitutively produce copious amounts of antibodies, imposing a critical risk factor for autoimmune disease. We previously found that plasma cell survival requires secreted factors such as APRIL and direct contact to stromal cells, which act in concert to activate NF-κB- and PI3K-dependent signaling pathways to prevent cell death. However, the regulatory properties of the underlying biochemical network are confounded by the complexity of potential interaction and cross-regulation pathways. Here, based on flow-cytometric quantification of key signaling proteins in the presence or absence of the survival signals APRIL and contact to the stromal cell line ST2, we generated a quantitative model of plasma cell survival. Our model emphasizes the non-redundant nature of the two plasma cell survival signals APRIL and stromal cell contact, and highlights a requirement for differential regulation of individual caspases. The modeling approach allowed us to unify distinct data sets and derive a consistent picture of the intertwined signaling and apoptosis pathways regulating plasma cell survival.
12
Mathias, Clinton B., Jeffrey Rovatti, and Stephanie Polukort. "IL-10 enhances IgE-independent IL-33-mediated mast cell cytokine production." Journal of Immunology 198, no. 1_Supplement (May 1, 2017): 145.8. http://dx.doi.org/10.4049/jimmunol.198.supp.145.8.
Full textAbstract:
Abstract IgE antibodies play a critical role in antigen-mediated mast cell activation. However, mast cells can be activated via IgE-independent mechanisms including signaling via the IL-33 receptor ST2. We have previously shown that the Th2-derived cytokine IL-10 can enhance IgE-dependent mucosal mast cell activation and promote mast cell function and survival. However, whether the effects of IL-10 on mast cells are global in nature and not limited to IgE-mediated signaling is not clear. To determine whether IL-10 can prime the activation of mast cells mediated by IgE-independent signals such as IL-33, we assessed the effects of rIL-10 exposure on IL-33-stimulated mast cells and their subsequent activation via IgE. Bone marrow-derived mast cells (BMMCs) from wild-type (WT) and IL-10−/− mice were cultured with rIL-33 with or without rIL-10 and activated with DNP-IgE and antigen. Exposure to IL-10 enhanced cytokine production in both WT and IL-10−/− BMMCs, and this was significantly increased after activation via IgE. IL-10−/−BMMCs exhibited decreased cytokine production, but this was restored upon culture with rIL-10. Interestingly, IL-33 stimulation (and subsequent activation with IgE) induced comparably enhanced levels of cytokine production, specifically IL-13 and IL-6, in both WT and IL-10−/−cells, suggesting that ST2-mediated signals can overcome the lack of IL-10. However, the addition of exogenous IL-10 to IL-33-stimulated cultures further enhanced the levels of these cytokines not only in IgE-activated cells, but also in cells stimulated with IL-33 alone. Our observations therefore suggest that IL-10 can play critical roles in enhancing mast cell function both via IgE-dependent and independent mechanisms.
13
Morales, Crystal, Joseph Maxwell, Adam Adler, and Anthony Vella. "Utilizing dual-costimulation through CD134 and CD137 with varying levels of antigen presentation to elicit heterogeneous CD4+ T cell responses (IRC9P.703)." Journal of Immunology 192, no. 1_Supplement (May 1, 2014): 191.4. http://dx.doi.org/10.4049/jimmunol.192.supp.191.4.
Full textAbstract:
Abstract Many cancer vaccines target CD8 T cells, but cytotoxic CD4+ T cells are emerging as important players. Our lab utilized a dual-costimulation immunotherapy approach through CD134 (OX40) and CD127 (4-1BB) agonist antibodies together with T cells specific to a widespread antigen which induced cytotoxic CD4+ T cell differentiation. Drawing upon a novel transgenic mouse model where CD11c+ cells present a specific antigen, we now demonstrate that antigen-specific dual-costimulated CD4+ T cells can be heterogeneous with differential expression of Eomes, KLRG1, GzmB, and the IL-33R (ST2). Specifically, in one transgenic mouse line with lower levels of CD11c antigen presentation, CD4+ T cells developed into Eomes+KLRG1+GzmB- cells. Despite comparable costimulation however, CD4+ T cells developed into Eomes-KLRG1-GzmB+ cells in a mouse line with greater levels of CD11c antigen presentation. Analysis of known helper T cell transcription factors and cytokines also showed expression of other helper T cell subset markers in these cells. Further, the capacity of these cells to become activated and kill target cells is currently being investigated with an in vitro assay. These findings are novel, demonstrating that cytotoxic CD4+ T cells may in fact be a heterogeneous population influenced by antigen presentation levels. Thus, this data should greatly impact immunotherapeutic targeting of cytotoxic CD4+ T cells.
14
Tkachev, Victor, Scott N. Furlan, E. Lake Potter, Hengqi Zheng, Daniel J. Hunt, Lucrezia Colonna, Agne Taraseviciute, et al. "Uncovering the Molecular Signature of Pathogenic Tissue-Infiltrating T Cells during Acute Graft-Versus-Host Disease." Blood 132, Supplement 1 (November 29, 2018): 805. http://dx.doi.org/10.1182/blood-2018-99-113652.
Full textAbstract:
Abstract One of the major barriers to developing targeted therapies for aGVHD control is the difficulty in identifying T cell signatures specific for GVHD pathology while distinguishing these from the pathways essential for tissue-specific T cell immune reconstitution. To address this need we have interrogated the migration patterns, as well as the phenotypic and transcriptomic characteristics of allogeneic T cells infiltrating aGVHD target organs in non-human primates. To rigorously study T cell migration during aGVHD we tracked T cells labeled following in vivo infusion with fluorescently tagged anti-CD45 antibodies given to NHP transplant recipients with aGVHD on day 8 post-HCT, during active disease. Anti-CD45 antibodies with distinct fluorescent tags were given at 6 hours before necropsy (anti-CD45-AlexaFluor647) and 5 minutes before necropsy (anti-CD45-AlexaFluor488), in order to measure T cells that were in the circulation and those migrating to GVHD target tissues, based on their labeling with one or both fluorescently-tagged antibodies. These experiments identified increased migration of both allogeneic CD8 T cells (Figure 1A) and CD4 T cells (not shown) during aGVHD, with trafficking into secondary lymphoid organs as well as non-lymphoid GVHD target organs (intestine and kidney). While migration was increased during aGVHD, these T cells, which demonstrated some phenotypic similarities to CD8 T cells in the peripheral blood (Figure 1B), also adopted tissue-specific phenotypes as measured by flow cytometry (Figure 1C), including the expression of canonical markers of resident-memory T cells (CD69+CD103-/+). However, unlike the tissue-resident T cells in healthy controls during homeostasis, tissue-infiltrating T cells during aGVHD expressed multiple markers of activation, including Ki67 and Granzyme B (Figure 1D). These flow cytometric characteristics suggested that the phenotype of organ-infiltrating T cells during aGVHD included attributes of both tissue-residency and of pathogenic alloreactivity. To further identify aGVHD-specific signatures, we performed transcriptomic analysis of tissue-infiltrating T cells during aGVHD. Using an unsupervised weighted gene correlation network analysis (WGCNA) we characterized the gene sets associated with individual GVHD target organs (Figure 2). We found that tissue-infiltrating T cells during aGVHD could be characterized by divergent features: First, they maintained a core tissue localization signature, which included genes previously linked to tissue-resident T cells (e.g. RUNX3, IFNG, CXCR6). Importantly, however, they also acquired an aGVHD-specific transcriptional signature including expression of IL1RL1 (encoding ST2), ICOS, TNFRSF9 (CD137) and TNFRSF4 (OX40). This signature also included enrichment for transcripts encoding the cytotoxic mediators GRMB and GRMA, the proliferation markers MKI67 and AURKA, as well as cytokines and cytokine receptors (IL18, IL18R, IL21, IL21R). Proteins encoded by each of these transcripts have been linked to aGVHD-causing T cells, strengthening the inference that these constitute a robust transcriptomic signature of aGVHD pathogenesis. Thus, for the first time in a large-animal model, we have been able to directly measure both the kinetics and the protein and RNA expression signatures of T cells during their migration into aGVHD target organs, This study provides new evidence for the evolution of a phenotypic and transcriptomic dichotomy during aGVHD-mediated tissue infiltration, in which T cells take on both tissue- and aGVHD-specific characteristics. These data provide novel insights into the spatial organization of systemic alloimmunity during aGVHD, which should enable more precise targeting of pathogenic T cell populations while preserving normal tissue immune reconstitution after transplantation. Disclosures Tkachev: Regeneron Pharmaceuticals, Inc.: Research Funding. Blazar:Kadmon Corporation, LLC: Consultancy, Research Funding. Kean:Regeneron Pharmaceuticals, Inc.: Research Funding.
15
Human, Christin, Sylvia Borchers, Michael Stadler, Helmut Diedrich, Jürgen Krauter, Christine Falk, Arnold Ganser, and Eva M. Mischak-Weissinger. "Investigation of Urine/Plasma Biomarkers to Predict Tolerance and Graft-Versus-Host Disease Post Allogeneic Stem Cell Transplantation." Blood 120, no. 21 (November 16, 2012): 3049. http://dx.doi.org/10.1182/blood.v120.21.3049.3049.
Full textAbstract:
Abstract Abstract 3049 Introduction Graft-versus-host-disease (GvHD) remains one of the major complications post allogeneic hematopoietic stem cell transplantation (HSCT) which is still the only curative treatment for hematologic malignancies and non-malignant hematopoietic disorders. Early detection of developing GvHD prior to clinical manifestation is in the focus of research in order to optimize therapeutic approaches and possibly lower transplant related morbidity and mortality. We previously published proteomic patterns in urine generated by capillary electrophoresis coupled on-line to mass spectrometry (CE-MS) allowing early detection of acute GvHD onset (Weissinger et al., 2007; Weissinger et al., 2009). In an effort to compare our CE-MS results to other proteomic approaches and to contribute to harmonization of results generated by different laboratories, we analyze biomarkers detected in urine as well as those described by others using proteomic approaches such as ELISA and Bio-Luminex in plasma collected from the same patients analyzed by CE-MS. We aim to (1) compare the predictive value of the proteomic pattern with other methods and (2) evaluate these methods for early diagnosis of GvHD. Methods We established an urine/plasma sample bank from patients transplanted since 2006 at MHH; of this four patient groups were chosen to set up the methods for monitoring emerging GvHD on the basis of plasma-biomarkers: patients at onset of aGvHD (aGvHD, n=20), patients at similar time-points early post HSCT without aGvHD (con_aGvHD, n=14), patients at diagnosis of cGvHD (cGvHD, n=13) and patients without GvHD (>day +100; con_cGvHD, n=33). For Bio-Luminex analysis, the angiogenesis marker panel was used (Luft et al., 2011), including: G-CSF, PECAM-1, HGF, VEGF, Leptin, PDGF-BB, Angiopoetin, Follistatin and IL-8. ELISAs were performed for the following previously identified markers: IL-2Rα, sTNFR1, ST2/IL-1R4, Reg3α, Semaphorin 5a, Elafin (Levine et al., 2012), CD99 and β2-microglobulin (CE-MS-analysis, Weissinger et al., submitted). Results & Discussion Within the training set analysis by Bio-Luminex no significant differences between aGvHD, cGvHD and respective controls was found for G-CSF, VEGF and PDGF, implying vascular processes due to immunosuppressive antibodies or toxicities early post-HSCT. Changes observed in the aGvHD group were more prominent when compared to the con_cGvHD group. However, this might reflect the completion of immunosuppression in most patients in the con_cGvHD-group. Interestingly, Leptin was significantly higher in patients w/o aGvHD shortly after HSCT compared to patients with aGvHD, but was also low in the cGvHD and con_cGvHD groups. For angiopoetin significantly different levels were found comparing aGvHD vs. con_aGvHD, as well as for cGvHD vs. con_cGvHD. Analysis of the training set showed a trend for angiopoetin distinguishing between patients with aGvHD and tolerant patients. Within the investigated markers of the ELISA analysis, CD99, TNFR1, IL-2Rα, and ST2/IL-1R4 yielded significant differences between the aGvHD and the control groups. First results for patients in the training set are shown in Figure 1. As the detected plasma concentration for these markers did not differ significantly in con_aGvHD and con_cGvHD, the control groups were merged. Preliminary analyses suggest that these markers might be suitable to distinguish patients with aGvHD from tolerant patients, as confirmed by receiver operated characteristics (ROC)-curves. Conclusions and future perspectives According to our preliminary results, the most promising markers to detect aGvHD in plasma samples are Leptin, Angiopoetin, TNFR1, β2-microglobulin, and IL-2Rα. In order to improve the predictive power of the ELISA and Bio-Luminex testing, a multiparametric model (support vector machines) will be applied to the training set data. Based on the training set, prospective screening of a validation set of patients will take place soon. Furthermore, by sequencing further relevant peptides from the urine proteomic pattern, we aim to identify additional biomarkers for GvHD. Disclosures: No relevant conflicts of interest to declare.
16
Lia, Giuseppe, Clara Di Vito, Marta Tapparo, Stefania Bruno, Elisa Zaghi, Michela Calvi, Lucia Brunello, et al. "Plasmatic Extracellular Vesicles in Acute Graft-Versus-Host Disease after Haplo-Identical Allografting with Post-Transplant Cyclophosphamide." Blood 134, Supplement_1 (November 13, 2019): 598. http://dx.doi.org/10.1182/blood-2019-126521.
Full textAbstract:
INTRODUCTION: Acute Graft-versus-Host-Disease (aGVHD) is a frequent complication where the endothelium may play a pivotal role. We recently investigated the potential role of extracellular vesicles (EVs) as novel biomarkers of aGVHD (Lia G. et al. Leukemia 2018). In this study we further investigated the correlation of plasma EVs and their content in miRNAs with the risk of developing aGVHD in the setting of post-transplant cyclophosphamide (PTCY) haploidentical-stem cell transplantation (Haplo-SCT). METHODS: Thirty-two patients who underwent a Haplo-SCT were included. Plasma samples were collected from peripheral blood at given time-points (pre-transplant, on day 0, 3, 7, 14, 21, 28, 35, 45, 60, 75 and 90 after transplant). EVs were extracted by a protamine-based precipitation method and were characterized by Nano-tracking Particle Analysis (Nanosight). EVs were then analyzed by flow-cytometry (Guava EasyCyte Flow Cytometer) with a panel of 14 antibodies (CD44, CD138, CD146, KRT18, CD120a, CD8, CD30, CD106, CD25, CD26, CD31, CD144, CD86, and CD140a). MiRNAs were extracted from EVs by miRNeasy Mini Kit (Qiagen) and retrotranscribed by miScript II RT Kit (Qiagen). Three miRNAs (miR100, miR194, miR155) were studied and quantified by qRT-PCR using the miScript SYBR Green PCR Kit (Qiagen). Concomitant plasma concentrations of human Tumor Necrosis Factor Receptor I (TNFR1) and human ST2 were also evaluated using a commercially available sandwich enzyme-linked immunosorbent assay (DualSET® ELISA R&D Systems). The risk of aGVHD was evaluated by logistic regression models and Odds Ratios (ORs) were estimated as absolute levels and as proportional changes compared with pre-transplant baseline levels of each marker. Moreover, among biomarkers significantly associated with a higher risk of aGVHD, a multivariable logistic regression model using Akaike's information criteria (AIC) was estimated to define a biomarker combination. Ors were reported for 1-unit increase of standardized variables. RESULTS: AGVHD (grade II-IV) was observed in 7/32 patients (22%) at a median of 41 (range 33-90) days after transplant. Logistic regression models showed that CD146 fluorescence was associated with a significantly increased risk of acute GVHD (OR 2.93 p<0.001) as well as expression changes in miR100, miR155 and miR194 (OR 3.90 p<0.001; OR 1.84 p=0.008; OR 2.68 p<0.001, respectively). Concentrations of plasmatic hTNFR1 and ST2 were also confirmed to be associated with increased risk of aGVHD (OR 1.47 p=0.04; OR 1.55 p=0.05, respectively) as previously described. Of note, all biomarkers associated with risk of aGVHD showed a consensual change in signal levels before the onset of aGVHD (Figure 1). By applying a backward selection on a multivariable logistic model using the AICapproach, we found that the combination of CD146-miR100-TNFR1 with an individual AUROC of 0.858, 0.923, and 0.794, respectively, increased their discrimination ability to predict aGVHD (multivariable AUROC = 0.987). CONCLUSIONS: This study, in the setting of haplo-transplant, confirms the association of CD146, a cell adhesion molecule, and the risk of aGVHD suggesting an important role of endothelium damage in the pathogenesis of aGVHD. The association of miRNA100, miRNA155 and miRNA194, carried by EVs, and aGVHD was also significant. Interestingly, MiRNA100 was reported to regulate inflammatory neovascularization during GvHD while miR-155 plays a role in donor T cell expansion. We have also found that using three markers in combination (CD146-miR100-TNFR1) could greatly improve aGVHD predictivity. To translate our results into an in vivo model, we have recently designed preclinical mouse models to evaluate if a) antagomir (against miRNA100 and/or miRNA155) injections or b) pre-emptive treatments with endothelium protective agents such as defibrinotide or OMS721 (Anti-Masp2) may reduce the risk of aGVHD. Figure1 a) Signal variation from baseline level (preTx) of CD146 fluorescence, miR100 expression, and TNFR1 concentration before aGVHD onset. Disclosures Boccadoro: Sanofi: Honoraria, Research Funding; Celgene: Honoraria, Research Funding; Amgen: Honoraria, Research Funding; Janssen: Honoraria, Research Funding; Novartis: Honoraria, Research Funding; AbbVie: Honoraria; Mundipharma: Research Funding; Bristol-Myers Squibb: Honoraria, Research Funding.
17
Neubauer, Heidi A., and Stuart M. Pitson. "Validation of commercially available sphingosine kinase 2 antibodies for use in immunoblotting, immunoprecipitation and immunofluorescence." F1000Research 5 (December 6, 2016): 2825. http://dx.doi.org/10.12688/f1000research.10336.1.
Full textAbstract:
Sphingosine kinase 2 (SK2) is a ubiquitously expressed lipid kinase that has important, albeit complex and poorly understood, roles in regulating cell survival and cell death. In addition to being able to promote cell cycle arrest and apoptosis under certain conditions, it has recently been shown that SK2 can promote neoplastic transformation and tumorigenesis in vivo. Therefore, well validated and reliable tools are required to study and better understand the true functions of SK2. Here, we compare two commercially available SK2 antibodies: a rabbit polyclonal antibody from Proteintech that recognizes amino acids 266-618 of human SK2a, and a rabbit polyclonal antibody from ECM Biosciences that recognizes amino acids 36-52 of human SK2a. We examine the performance of these antibodies for use in immunoblotting, immunoprecipitation and immunofluorescence staining of endogenous SK2, using human HEK293 and HeLa cell lines, as well as mouse embryonic fibroblasts (MEFs). Furthermore, we assess the specificity of these antibodies to the target protein through the use of siRNA-mediated SK2 knockdown and SK2 knockout (Sphk2-/-) MEFs. Our results demonstrate that the Proteintech anti-SK2 antibody reproducibly displayed superior sensitivity and selectivity towards SK2 in immunoblot analyses, while the ECM Biosciences anti-SK2 antibody was reproducibly superior for SK2 immunoprecipitation and detection by immunofluorescence staining. Notably, both antibodies produced non-specific bands and staining in the MEFs, which was not observed with the human cell lines. Therefore, we conclude that the Proteintech SK2 antibody is a valuable reagent for use in immunoblot analyses, and the ECM Biosciences SK2 antibody is a useful tool for SK2 immunoprecipitation and immunofluorescence staining, at least in the human cell lines employed in this study.
18
Neubauer, Heidi A., and Stuart M. Pitson. "Validation of commercially available sphingosine kinase 2 antibodies for use in immunoblotting, immunoprecipitation and immunofluorescence." F1000Research 5 (March 23, 2017): 2825. http://dx.doi.org/10.12688/f1000research.10336.2.
Full textAbstract:
Sphingosine kinase 2 (SK2) is a ubiquitously expressed lipid kinase that has important, albeit complex and poorly understood, roles in regulating cell survival and cell death. In addition to being able to promote cell cycle arrest and apoptosis under certain conditions, it has recently been shown that SK2 can promote neoplastic transformation and tumorigenesis in vivo. Therefore, well validated and reliable tools are required to study and better understand the true functions of SK2. Here, we compare two commercially available SK2 antibodies: a rabbit polyclonal antibody from Proteintech that recognizes amino acids 266-618 of human SK2a, and a rabbit polyclonal antibody from ECM Biosciences that recognizes amino acids 36-52 of human SK2a. We examine the performance of these antibodies for use in immunoblotting, immunoprecipitation and immunofluorescence staining of endogenous SK2, using human HEK293 and HeLa cell lines, as well as mouse embryonic fibroblasts (MEFs). Furthermore, we assess the specificity of these antibodies to the target protein through the use of siRNA-mediated SK2 knockdown and SK2 knockout (Sphk2-/-) MEFs. Our results demonstrate that the Proteintech anti-SK2 antibody reproducibly displayed superior sensitivity and selectivity towards SK2 in immunoblot analyses, while the ECM Biosciences anti-SK2 antibody was reproducibly superior for SK2 immunoprecipitation and detection by immunofluorescence staining. Notably, both antibodies produced non-specific bands and staining in the MEFs, which was not observed with the human cell lines. Therefore, we conclude that the Proteintech SK2 antibody is a valuable reagent for use in immunoblot analyses, and the ECM Biosciences SK2 antibody is a useful tool for SK2 immunoprecipitation and immunofluorescence staining, at least in the human cell lines employed in this study.
19
Ogega, Clinton O., Nicole E. Skinner, Andrew I. Flyak, Kaitlyn E. Clark, Nathan L. Board, Pamela J. Bjorkman, James E. Crowe, Andrea L. Cox, Stuart C. Ray, and Justin R. Bailey. "B cell overexpression of FCRL5 and PD-1 is associated with low antibody titers in HCV infection." PLOS Pathogens 18, no. 1 (January 6, 2022): e1010179. http://dx.doi.org/10.1371/journal.ppat.1010179.
Full textAbstract:
Antibodies targeting the hepatitis C virus (HCV) envelope glycoprotein E2 are associated with delayed disease progression, and these antibodies can also facilitate spontaneous clearance of infection in some individuals. However, many infected people demonstrate low titer and delayed anti-E2 antibody responses. Since a goal of HCV vaccine development is induction of high titers of anti-E2 antibodies, it is important to define the mechanisms underlying these suboptimal antibody responses. By staining lymphocytes with a cocktail of soluble E2 (sE2) glycoproteins, we detected HCV E2-specific (sE2+) B cells directly ex vivo at multiple acute infection timepoints in 29 HCV-infected subjects with a wide range of anti-E2 IgG titers, including 17 persistently infected subjects and 12 subjects with spontaneous clearance of infection. We performed multi-dimensional flow cytometric analysis of sE2+ and E2-nonspecific (sE2-) class-switched B cells (csBC). In sE2+ csBC from both persistence and clearance subjects, frequencies of resting memory B cells (rMBC) were reduced, frequencies of activated MBC (actMBC) and tissue-like MBC (tlMBC) were increased, and expression of FCRL5, an IgG receptor, was significantly upregulated. Across all subjects, plasma anti-E2 IgG levels were positively correlated with frequencies of sE2+ rMBC and sE2+ actMBC, while anti-E2 IgG levels were negatively correlated with levels of FCRL5 expression on sE2+ rMBC and PD-1 expression on sE2+ actMBC. Upregulation of FCRL5 on sE2+ rMBC and upregulation of PD-1 on sE2+ actMBC may limit anti-E2 antibody production in vivo. Strategies that limit upregulation of these molecules could potentially generate higher titers of protective antibodies against HCV or other pathogens.
20
González-Rubio, Gema, Ángela Sellers-Moya, Humberto Martín, and María Molina. "Differential Role of Threonine and Tyrosine Phosphorylation in the Activation and Activity of the Yeast MAPK Slt2." International Journal of Molecular Sciences 22, no. 3 (January 23, 2021): 1110. http://dx.doi.org/10.3390/ijms22031110.
Full textAbstract:
The Mitogen-Activated Protein Kinase (MAPK) Slt2 is central to signaling through the yeast Cell Wall Integrity (CWI) pathway. MAPKs are regulated by phosphorylation at both the threonine and tyrosine of the conserved TXY motif within the activation loop (T190/Y192 in Slt2). Since phosphorylation at both sites results in the full activation of MAPKs, signaling through MAPK pathways is monitored with antibodies that detect dually phosphorylated forms. However, most of these antibodies also recognize monophosphorylated species, whose relative abundance and functionality are diverse. By using different phosphospecific antibodies and phosphate-affinity (Phos-tag) analysis on distinct Slt2 mutants, we determined that Y192- and T190-monophosphorylated species coexist with biphosphorylated Slt2, although most of the Slt2 pool remains unphosphorylated following stress. Among the monophosphorylated forms, only T190 exhibited biological activity. Upon stimulation, Slt2 is first phosphorylated at Y192, mainly by the MAPKK Mkk1, and this phosphorylation is important for the subsequent T190 phosphorylation. Similarly, dephosphorylation of Slt2 by the Dual Specificity Phosphatase (DSP) Msg5 is ordered, with dephosphorylation of T190 depending on previous Y192 dephosphorylation. Whereas Y192 phosphorylation enhances the Slt2 catalytic activity, T190 is essential for this activity. The conserved T195 residue is also critical for Slt2 functionality. Mutations that abolish the activity of Slt2 result in a high increase in inactive Y192-monophosphorylated Slt2. The coexistence of different Slt2 phosphoforms with diverse biological significance highlights the importance of the precise detection of the Slt2 phosphorylation status.
21
Siddiqui, Asim A., Jia Xainli, Jesse Schloegel, Lenore Carias, Francis Ntumngia, Menachem Shoham, Joanne L. Casey, Michael Foley, John H. Adams, and Christopher L. King. "Fine Specificity of Plasmodium vivax Duffy Binding Protein Binding Engagement of the Duffy Antigen on Human Erythrocytes." Infection and Immunity 80, no. 8 (May 21, 2012): 2920–28. http://dx.doi.org/10.1128/iai.00206-12.
Full textAbstract:
ABSTRACTPlasmodium vivaxinvasion of human erythrocytes requires interaction of theP. vivaxDuffy binding protein (PvDBP) with its host receptor, the Duffy antigen (Fy) on the erythrocyte surface. Consequently, PvDBP is a leading vaccine candidate. The binding domain of PvDBP lies in a cysteine-rich portion of the molecule called region II (PvDBPII). PvDBPII contains three distinct subdomains based upon intramolecular disulfide bonding patterns. Subdomain 2 (SD2) is highly polymorphic and is thought to contain many key residues for binding to Fy, while SD1 and SD3 are comparatively conserved and their role in Fy binding is not well understood. To examine the relative contributions of the different subdomains to binding to Fy and their abilities to elicit strain-transcending binding-inhibitory antibodies, we evaluated recombinant proteins from SD1+2, SD2, SD3, and SD3+, which includes 24 residues of SD2. All of the recombinant subdomains, except for SD2, bound variably to human erythrocytes, with constructs containing SD3 showing the best binding. Antisera raised in laboratory animals against SD3, SD3+, and SD2+3 inhibited the binding of full-length PvDBPII, which is strain transcending, whereas antisera generated to SD1+2 and SD2 failed to generate blocking antibodies. All of the murine monoclonal antibodies generated to full-length PvDBPII that had significant binding-inhibitory activity recognized only SD3. Thus, SD3 binds Fy and elicits blocking antibodies, indicating that it contains residues critical to Fy binding that could be the basis of a strain-transcending candidate vaccine againstP. vivax.
22
Tatzber, Franz, Edith Pursch, Ulrike Resch, Roswitha Pfragner, Sandra Holasek, Meinrad Lindschinger, Gerhard Cvirn, and Willibald Wonisch. "Cultivation and Immortalization of Human B-Cells Producing a Human Monoclonal IgM Antibody Binding to MDA-LDL: Further Evidence for Formation of Atherogenic MDA-LDL Adducts in HumansIn Vivo." Oxidative Medicine and Cellular Longevity 2017 (2017): 1–7. http://dx.doi.org/10.1155/2017/6047142.
Full textAbstract:
Oxidatively modified low-density lipoprotein (oLDL) is firmly believed to play an important role in the initiation and development of atherosclerosis, and malonic dialdehyde (MDA) is one of the major lipid peroxidation breakdown products involved in this process. In recent decades, antibodies against MDA-LDL have been detected in human and animal sera. In our study, human B-cells from the peripheral blood of a healthy female donor were fused with the SP2/0 mouse myeloma cell line. Antibody-producing hybridomas were detected by MDA-LDL-IgG/IgM enzyme-linked immunosorbent assays (ELISA) and Cu++-oxidized LDL IgG/IgM (oLAb) ELISA. Cells with supernatants emitting positive signals for antibodies were then cloned and after sufficient multiplication frozen and stored under liquid nitrogen. Due to the loss of antibody-producing ability, we established an MDA-LDL-IgM-producing cell line by recloning. This allowed isolation and immortalization of several human B-cells. The human donor had not been immunized with MDA-modified proteins, thus obviously producing MDA-LDL antibodiesin vivo. Furthermore, using these antibodies forin vitroexperiments, we were able to demonstrate that MDA epitopes are among the epitopes generated during Cu++-LDL oxidation as well. Finally, these antibodies compete in ELISA and cell culture experiments with MDA as a challenging toxin or ligand.
23
Forsey, T. "Antibodies to Chlamydia trachomatis." Sexually Transmitted Infections 63, no. 4 (August 1, 1987): 279. http://dx.doi.org/10.1136/sti.63.4.279.
Full text
24
Bhattacharya-Chatterjee, M., M. W. Pride, B. K. Seon, and H. Kohler. "Idiotype vaccines against human T cell acute lymphoblastic leukemia. I. Generation and characterization of biologically active monoclonal anti-idiotopes." Journal of Immunology 139, no. 4 (August 15, 1987): 1354–60. http://dx.doi.org/10.4049/jimmunol.139.4.1354.
Full textAbstract:
Abstract A murine monoclonal anti-tumor antibody termed SN2 (Ab1), isotype IgG1-kappa, that defines a unique human T cell leukemia-associated cell-surface glycoprotein, gp37 (m.w. 37,000), was used to generate monoclonal anti-idiotype antibodies (Ab2) in syngeneic BALB/c mice. The Ab2 were screened on the basis of their binding to the F(ab')2 fragments of SN2 and not to the F(ab')2 of pooled normal BALB/c mice sera IgG1 or to an unrelated BALB/c monoclonal antibody of the same isotype. Fifteen Ab2, obtained from two fusions, were specific for the SN2 idiotope and not against isotype or allotype determinants. To find out whether these Ab2 are directed against the paratope of SN2, the binding of radiolabeled SN2 to leukemic MOLT-4 and JM cells which contain gp37 as a surface constituent was studied in the presence of these anti-idiotopes. Clone 4EA2 inhibited the binding 100% at a concentration of 50 ng and 4DC6 inhibited 90% at a concentration of 250 ng. A third clone 4DD6 gave about 50% inhibition. Similar was the inhibition of SN2 binding to insolubilized MOLT-4 antigen or cell membrane preparation. The binding of SN2 (Ab1) to 4EA2 and 4DC6 was also inhibited by semipurified preparation of gp37 antigen. These results demonstrate that at least two of the anti-idiotope antibodies are binding either at or near the binding site idiotope of SN2. Next, the purified Ab2 was used to immunize syngeneic mice to induce antibody binding to MOLT-4 cells or gp37. Sera from mice immunized with 4EA2 and 4DC6 coupled to keyhole limpet hemocyanin contained antibodies which bind to semipurified gp37 antigen and MOLT-4 cells. Immune sera inhibited the binding of iodinated Ab2 and Ab1 indicating that an anti-anti-idiotopic antibody (Ab3) in mice shares idiotopes with Ab1 (SN2). Also, the binding of iodinated Ab2 to Ab1 was inhibited by rabbit antisera specific for gp37. Collectively, these data suggest that anti-idiotype antibodies 4EA2 and 4DC6 may be useful in the generation of idiotype vaccines against human T cell leukemia.
25
Angelelli, C., K. Stefanantoni, M. Cadar, G. Pellegrino, F. Conti, and V. Riccieri. "POS0433 CAN INTERLEUKIN 33 (IL-33) BE CONSIDERED A VALUABLE BIOMARKER IN THE EARLY STAGES OF SYSTEMIC SCLEROSIS? ANALYSIS OF A MONOCENTRIC COHORT." Annals of the Rheumatic Diseases 80, Suppl 1 (May 19, 2021): 445.2–445. http://dx.doi.org/10.1136/annrheumdis-2021-eular.4196.
Full textAbstract:
Background:The ScS approach has changed considerably in recent years especially concerning the very early diagnosis of the disease (VEDOSS) at the time when the patient is still in an undifferentiated form (UCTD) at risk of developing SSc (1, 2). Of great value are different clinical, instrumental and laboratory findings, such as specific autoantibodies and Nailfold VideoCapillaroscopy (NVC), able to identify those cases progressing into overt SSc. IL-33 cytokine is known to exert pro-fibrotic effects through its membrane receptor ST2 on immune cells and myofibroblasts and recent studies suggest that it can be released following endothelial cell activation at the onset of SSc (3, 4).Objectives:Our aim has been to evaluate IL-33 serum levels in a monocentric cohort of VEDOSS patients, looking for the possible association with clinical phenotype and disease progression, focusing on the microvascular capillaroscopic changes.Methods:Fourty-seven VEDOSS patients underwent a complete clinical, instrumental and laboratory evaluation, including NVC and specific SSc autoantibodies. At baseline serum IL-33 levels were measured using an ELISA assay. In 32 of them we also had a second serum sample at a follow-up time of at least 24 months (range 24 to 96 months).Results:During the follow-up time, 17 patients were subsequently reclassified as having ScS whereas 30 remained VEDOSS. The “progressor” subjects positively correlated with the presence of anti-Topoisomerase I antibodies (p>0,004). IL-33 concentrations had a median value of 427.2 pg/ml (IQR 967.9 pg/ml) at baseline and of 130.4 pg/ml (IQR 399 pg/ml) at the follow up, showing a statistically significant difference independently from the progression of the disease (p=0.03). Besides significantly higher levels were detected in those patients with more severe NVC changes, defined as “active” pattern (p<0.05). Among the 47 VEDOSS patients, 12 started some kind of vascular therapy. In these patients serum IL-33 concentrations significantly lowered during the follow-up respect to those without any treatment (p<0.03)Conclusion:The analysis of our data confirms previous report (5) on higher IL-33 serum levels in the very early stages of UCTD patients at risk for SSc, regardless of their progression in established SSc, although related to more severe microvascular NVC involvement. The lowering of IL-33 serum levels that we detected in the follow up of our patients, may be linked to the well-known endothelial changes during the progression of the SSc and seems also to be partially affected by treatments. Investigation on a greater number of patients are needed to better understand our findings.References:[1]J. Avouac et al. Ann Rheum Dis 2011[2]G Valentini et al. Clin Exp Med 2017[3]Manetti M, et al. Ann Rheum Dis 2010[4]Terras S et al. Ann Rheum Dis 2013[5]Vettori S, et al. J Clin Immunol 2014Disclosure of Interests:None declared
26
Montalbán, J., J. Ordi, J. Barquinero, and M. Vilardell. "Sneddon's syndrome and anticardiolipin antibodies." Stroke 19, no. 6 (June 1988): 785–86. http://dx.doi.org/10.1161/str.19.6.3376175.
Full text
27
Ghosh, S. K., and T. N. Naik. "Evidence for a new rotavirus subgroup in India." Epidemiology and Infection 102, no. 3 (June 1989): 523–30. http://dx.doi.org/10.1017/s0950268800030235.
Full textAbstract:
SUMMARYMonoclonal antibodies specific for rotavirus subgroup 1 (SG1) and subgroup 2 (SG2) were used to analyse by enzyme immunoassay (EIA) the subgroups of human rotavirus isolates obtained from three different parts of India during the period September 1985 to July 1987. We identified one isolate which failed to react with either SG1 or SG2 specific monoclonal antibodies, although it reacted well with a monoclonal antibody specific for group A rotaviruses. This finding suggests that it belongs to a new rotavirus subgroup. Further, another isolate was found to belong to SG1 although it had a ‘long’ electropherotype.
28
Martinez-Menendez, B., A. Pérez-Sempere, M. Gonzalez-Rubio, F. J. Villaverde-Amundarain, and F. Bermejo-Pareja. "Sneddon's syndrome with negative antiphospholipid antibodies." Stroke 21, no. 10 (October 1990): 1510–11. http://dx.doi.org/10.1161/str.21.10.1510b.
Full text
29
Hering, R., E. G. Couturier, and T. J. Steiner. "Vascular headaches and the anticardiolipin antibodies." Stroke 22, no. 3 (March 1991): 414–15. http://dx.doi.org/10.1161/str.22.3.414b.
Full text
30
Li, Dapeng, Markus von Schaewen, Xuesong Wang, Wanyin Tao, Yunfang Zhang, Li Li, Brigitte Heller, et al. "Altered Glycosylation Patterns Increase Immunogenicity of a Subunit Hepatitis C Virus Vaccine, Inducing Neutralizing Antibodies Which Confer Protection in Mice." Journal of Virology 90, no. 23 (September 14, 2016): 10486–98. http://dx.doi.org/10.1128/jvi.01462-16.
Full textAbstract:
ABSTRACT Hepatitis C virus (HCV) infection is a global health problem for which no vaccine is available. HCV has a highly heterogeneous RNA genome and can be classified into seven genotypes. Due to the high genetic and resultant antigenic variation among the genotypes, inducing antibodies capable of neutralizing most of the HCV genotypes by experimental vaccination has been challenging. Previous efforts focused on priming humoral immune responses with recombinant HCV envelope E2 protein produced in mammalian cells. Here, we report that a soluble form of HCV E2 (sE2) produced in insect cells possesses different glycosylation patterns and is more immunogenic, as evidenced by the induction of higher titers of broadly neutralizing antibodies (bNAbs) against cell culture-derived HCV (HCVcc) harboring structural proteins from a diverse array of HCV genotypes. We affirm that continuous and discontinuous epitopes of well-characterized bNAbs are conserved, suggesting that sE2 produced in insect cells is properly folded. In a genetically humanized mouse model, active immunization with sE2 efficiently protected against challenge with a heterologous HCV genotype. These data not only demonstrate that sE2 is a promising HCV vaccine candidate, but also highlight the importance of glycosylation patterns in developing subunit viral vaccines. IMPORTANCE A prophylactic vaccine with high efficacy and low cost is urgently needed for global control of HCV infection. Induction of broadly neutralizing antibodies against most HCV genotypes has been challenging due to the antigenic diversity of the HCV genome. Here, we refined a high-yield subunit HCV vaccine that elicited broadly neutralizing antibody responses in preclinical trials. We found that soluble HCV E2 protein (sE2) produced in insect cells is distinctly glycosylated and is more immunogenic than sE2 produced in mammalian cells, suggesting that glycosylation patterns should be taken into consideration in efforts to generate antibody-based recombinant vaccines against HCV. We further showed that sE2 vaccination confers protection against HCV infection in a genetically humanized mouse model. Thus, our work identified a promising broadly protective HCV vaccine candidate that should be considered for further preclinical and clinical development.
31
Kaul, DK, RL Nagel, D. Chen, and HM Tsai. "Sickle erythrocyte-endothelial interactions in microcirculation: the role of von Willebrand factor and implications for vasoocclusion." Blood 81, no. 9 (May 1, 1993): 2429–38. http://dx.doi.org/10.1182/blood.v81.9.2429.2429.
Full textAbstract:
Abstract To determine the role of von Willebrand factor (vWF) in adhesion of sickle (SS) erythrocytes in microvascular flow conditions, we have perfused the ex vivo mesocecum vasculature of the rat with desmopressin, an analogue of vasopressin that causes the release of endothelial vWF. Analysis of vWF in the venous effluent of the isolated vasculature showed mainly the presence of extra-large molecular weight forms characteristic of endothelial vWF, which in the presence of desmopressin showed an average increase of 54%. Also, desmopressin induced a significant increase in adhesion of washed oxygenated (oxy) unseparated SS erythrocytes, accompanied by a persistent microvascular obstruction and a pronounced increase in the peripheral resistance (PRU). In contrast, infusion of SS deformable discocytes (SS2) in desmopressin-perfused vasculature resulted in a significant adhesion but not in persistent vasoocclusion, showing that SS2 discocytes alone are not sufficient for microvascular obstruction. Furthermore, SS4 erythrocytes (dense discocytes and irreversibly sickled erythrocytes) caused a persistent microvascular blockage and a significantly higher PRU than SS2 discocytes. However, the increase in PRU for SS4 erythrocytes following desmopressin treatment was 50% less compared with a corresponding increase for SS2 discocytes over the control values, which showed a smaller effect of desmopressin on the hemodynamic behavior of SS4 dense erythrocytes. Incubation of desmopressin-treated vasculature with anti-vWF antibodies resulted in a pronounced decrease in adhesion and significantly improved hemodynamic behavior of SS cells. Also, in untreated vasculature, similarly incubated with anti-vWF antibodies, there was almost complete inhibition of adhesion. Under the described perfusion conditions, antibodies to fibronectin and thrombospondin, as well as incubation of SS erythrocytes with anti-vWF antibodies did not affect adhesion. These results are compatible with a model for SS vasoocclusion in which extra- large vWF-mediated adhesion of deformable SS erythrocytes is the first step followed by an accelerated entrapment of dense SS erythrocytes.
32
Kaul, DK, RL Nagel, D. Chen, and HM Tsai. "Sickle erythrocyte-endothelial interactions in microcirculation: the role of von Willebrand factor and implications for vasoocclusion." Blood 81, no. 9 (May 1, 1993): 2429–38. http://dx.doi.org/10.1182/blood.v81.9.2429.bloodjournal8192429.
Full textAbstract:
To determine the role of von Willebrand factor (vWF) in adhesion of sickle (SS) erythrocytes in microvascular flow conditions, we have perfused the ex vivo mesocecum vasculature of the rat with desmopressin, an analogue of vasopressin that causes the release of endothelial vWF. Analysis of vWF in the venous effluent of the isolated vasculature showed mainly the presence of extra-large molecular weight forms characteristic of endothelial vWF, which in the presence of desmopressin showed an average increase of 54%. Also, desmopressin induced a significant increase in adhesion of washed oxygenated (oxy) unseparated SS erythrocytes, accompanied by a persistent microvascular obstruction and a pronounced increase in the peripheral resistance (PRU). In contrast, infusion of SS deformable discocytes (SS2) in desmopressin-perfused vasculature resulted in a significant adhesion but not in persistent vasoocclusion, showing that SS2 discocytes alone are not sufficient for microvascular obstruction. Furthermore, SS4 erythrocytes (dense discocytes and irreversibly sickled erythrocytes) caused a persistent microvascular blockage and a significantly higher PRU than SS2 discocytes. However, the increase in PRU for SS4 erythrocytes following desmopressin treatment was 50% less compared with a corresponding increase for SS2 discocytes over the control values, which showed a smaller effect of desmopressin on the hemodynamic behavior of SS4 dense erythrocytes. Incubation of desmopressin-treated vasculature with anti-vWF antibodies resulted in a pronounced decrease in adhesion and significantly improved hemodynamic behavior of SS cells. Also, in untreated vasculature, similarly incubated with anti-vWF antibodies, there was almost complete inhibition of adhesion. Under the described perfusion conditions, antibodies to fibronectin and thrombospondin, as well as incubation of SS erythrocytes with anti-vWF antibodies did not affect adhesion. These results are compatible with a model for SS vasoocclusion in which extra- large vWF-mediated adhesion of deformable SS erythrocytes is the first step followed by an accelerated entrapment of dense SS erythrocytes.
33
Estevez Garcia, Andrea Isabel, David J. Lefebvre, Lionel Nyabongo, Andy Haegeman, Canesius Nkundwanayo, Annebel De Vleeschauwer, Désiré Ntakirutimana, et al. "Outbreaks of Foot-and-Mouth Disease in Burundi, East Africa, in 2016, Caused by Different Serotypes." Viruses 14, no. 5 (May 17, 2022): 1077. http://dx.doi.org/10.3390/v14051077.
Full textAbstract:
Burundi is a small, densely populated country in the African Great Lakes region. In March 2016, several hundreds of cattle were reported with vesicular lesions, suggesting foot-and-mouth disease (FMD). Epithelial samples, saliva, and blood were collected in six of the affected provinces spread over the country. The overall seroprevalence of FMD virus (FMDV) in the affected herds, as determined by antibodies against FMDV non-structural proteins, was estimated at 87%. Antibodies against FMDV serotypes O (52%), A (44%), C (19%), SAT1 (36%), SAT2 (58%), and SAT3 (23%) were detected across the provinces. FMDV genome was detected in samples from five of the six provinces using rRT-PCR. FMDV was isolated from samples from three provinces: in Cibitoke province, serotypes A and SAT2 were isolated, while in Mwaro and Rutana provinces, only serotype SAT2 was isolated. In Bururi and Cankuzo provinces, the serological profile suggested a recent incursion with serotype SAT2, while in Bubanza province, the serological profile suggested past incursions with serotype O and possibly serotype SAT1. The phylogenetic assessments showed the presence of topotypes A/Africa/G-I and SAT2/IV, similarly to previously characterized virus strains from other countries in the region, suggesting a transboundary origin and necessitating a regional approach for vaccination and control of FMD.
34
Montalbán, J., A. Codina, J. Ordi, M. Vilardell, M. A. Khamashta, and G. R. Hughes. "Antiphospholipid antibodies in cerebral ischemia." Stroke 22, no. 6 (June 1991): 750–53. http://dx.doi.org/10.1161/01.str.22.6.750.
Full text
35
Schmidt, R., P. Auer-Grumbach, F. Fazekas, H. Offenbacher, and P. Kapeller. "Anticardiolipin Antibodies in Normal Subjects." Stroke 26, no. 5 (May 1995): 749–54. http://dx.doi.org/10.1161/01.str.26.5.749.
Full text
36
Luong, Thanh-Ha, Jacob H. Rand, Xiao-Xuan Wu, James H. Godbold, Mayra Gascon-Lema, and Stanley Tuhrim. "Seasonal Distribution of Antiphospholipid Antibodies." Stroke 32, no. 8 (August 2001): 1707–11. http://dx.doi.org/10.1161/01.str.32.8.1707.
Full text
37
Benhamou, Nicole, G. B. Ouellette, J. G. Lafontaine, and J. R. Joly. "Use of monoclonal antibodies to detect a phytotoxic glycopeptide produced by Ophiostoma ulmi, the Dutch elm disease pathogen." Canadian Journal of Botany 63, no. 7 (July 1, 1985): 1177–84. http://dx.doi.org/10.1139/b85-163.
Full textAbstract:
Two hybridomas that secrete antibodies specific for a phytotoxic glycopeptide from Ophiostoma ulmi (Buism.) Nannf. were produced by fusing spleen cells of mice immunized with the purified toxin and the Sp2-0/Ag14 mouse myeloma cell line. Specificity of these antibodies was first demonstrated by an enzyme-linked immunosorbent assay (ELISA), then by immunoblotting on nitrocellulose membrane after sodium dodecyl sulfate – polyacrylamide gel electrophoresis of the glycopeptide. Both clones produced antibodies of IgM class as determined by immunodiffusion. These monoclonal antibodies were utilized to detect and localize the toxic glycopeptide in pathogen cells and infected host tissues by immunohistochemical and immunocytochemical techniques.
38
Kuklina, G. V., S. S. Ipatov, D. V. Pechenkin, A. V. Eremkin, A. A. Kytmanov, M. Yu Dubrovin, and A. S. Gorshkov. "Obtaining and Characterization of Hybridomas Producing Monoclonal Antibodies to Shiga-Like Toxins of I and II Types." Problems of Particularly Dangerous Infections, no. 3 (October 23, 2021): 83–88. http://dx.doi.org/10.21055/0370-1069-2021-3-83-88.
Full textAbstract:
Objective – obtaining and characterization of hybrid cell lines producing monoclonal antibodies against I and II types of shiga-like toxins.Materials and methods. Shiga-like toxins obtained in “48thCentral Research Institute” of Ministry of Defense of Russian Federation (Kirov), BALB/c mice, myeloma cells SP2/0-Ag14 were used in research. Immune splenocytes and SP2/0-Ag14 myeloma cells were fused according to G. Kohler and C. Milstein method in De St. Fazekas and D. Scheidegger modifcation using 50 % polyethylene glycol. Hybrid cell lines producing specifc monoclonal antibodies were cloned by limited dilutions. Hybridomas growth and producing properties were studied in vitro and in vivo. Specifc activity of immune sera, culture and ascitic fluids were studied by indirect ELISA. Monoclonal antibodies from ascitic fluids were precipitated by saturated ammonium sulfate, followed by ion exchange chromatographyResults and discussion. 8 hybridomas producing monoclonal antibodies against I and II types shiga-like toxins were obtained. Hybridomas are characterized by stable proliferation and antibody-producing activity during 10 passages in vitro and 3 passages in vivo (observation period). Obtained monoclonal antibodies can be used for ELISA detection of I and II types shiga-like toxins. Minimum detectable concentration of shiga-like toxins in sandwich ELISA is 1 ng/ml. The possibility of detecting shiga-like toxins without typical differentiation was shown when using in the enzyme immunoassay a polyreceptor mixture of monoclonal antibodies for sensitizing the plate and a polyspecifc mixture of immunoperoxidase conjugates.
39
Beidler, C. B., J. R. Ludwig, J. Cardenas, J. Phelps, C. G. Papworth, E. Melcher, M. Sierzega, L. J. Myers, B. W. Unger, and M. Fisher. "Cloning and high level expression of a chimeric antibody with specificity for human carcinoembryonic antigen." Journal of Immunology 141, no. 11 (December 1, 1988): 4053–60. http://dx.doi.org/10.4049/jimmunol.141.11.4053.
Full textAbstract:
Abstract A mouse/human chimeric antibody has been constructed by using variable light and variable heavy regions from a murine hybridoma specific for human carcinoembryonic antigen (CEA) (CEM231.6.7). These V regions were combined with kappa and gamma-1 constant region genes cloned from human lymphocytes. The chimeric constructs were sequentially electroporated into murine non-Ig-producing myeloma (P3.653) and hybridoma (SP2/0) cell. Significant differences were seen in expression levels between the two cell types. High levels of expression (24 to 32 micrograms/ml/10(6) cells) were seen with several of the anti-CEA SP2/0 transfectomas but not with the P3.653 cells. The SP2/0 transfectoma lines were adapted to serum-free, chemically defined media and grown in large scale fermentation cultures where they continued to secrete high levels of antibody. The chimeric antibodies remain reactive against human CEA with affinity constants comparable to that of the parental hybridoma antibody. High level expression will make practical the production of chimeric antibodies for in vivo therapeutic and diagnostic purposes.
40
Levine, S. R., S. Kim, M. J. Deegan, and K. M. Welch. "Ischemic stroke associated with anticardiolipin antibodies." Stroke 18, no. 6 (November 1987): 1101–6. http://dx.doi.org/10.1161/01.str.18.6.1101.
Full text
41
Coull, B. M., and S. H. Goodnight. "Antiphospholipid antibodies, prethrombotic states, and stroke." Stroke 21, no. 9 (September 1990): 1370–74. http://dx.doi.org/10.1161/01.str.21.9.1370.
Full text
42
Carhuapoma, Juan R., Panayiotis Mitsias, and Steven R. Levine. "Cerebral Venous Thrombosis and Anticardiolipin Antibodies." Stroke 28, no. 12 (December 1997): 2363–69. http://dx.doi.org/10.1161/01.str.28.12.2363.
Full text
43
Tanne, David, Douglas A. Triplett, and Steven R. Levine. "Antiphospholipid-Protein Antibodies and Ischemic Stroke." Stroke 29, no. 9 (September 1998): 1755–58. http://dx.doi.org/10.1161/01.str.29.9.1755.
Full text
44
McCartney, A. C., J. L. Winning, and I. B. Tait. "Monoclonal antibodies in identifying Neisseria gonorrhoeae: cautionary note." Sexually Transmitted Infections 63, no. 6 (December 1, 1987): 398. http://dx.doi.org/10.1136/sti.63.6.398.
Full text
45
Shattil, SJ, M. Cunningham, and JA Hoxie. "Detection of activated platelets in whole blood using activation- dependent monoclonal antibodies and flow cytometry." Blood 70, no. 1 (July 1, 1987): 307–15. http://dx.doi.org/10.1182/blood.v70.1.307.307.
Full textAbstract:
Abstract Platelets may become activated in a number of clinical disorders and participate in thrombus formation. We developed a direct test for activated platelets in whole blood using flow cytometry. Whole blood was incubated with either biotin-PAC1, a monoclonal antibody specific for the fibrinogen receptor on activated platelets, or biotin-S12, an antibody specific for an alpha-granule membrane protein that associates with the platelet surface during secretion. Platelet-bound antibodies were detected with streptavidin conjugated with fluorescein isothiocyanate (FITC) or phycoerythrin (PE). Platelets were differentiated from the larger erythrocytes and WBCs by their light- scatter profile. Alternatively, platelets could be identified with FITC- AP1, an antibody specific for platelet membrane glycoprotein Ib, and analyzed further for PAC1 or S12 binding with PE-streptavidin. No centrifugation or washing steps were required. With gel-filtered platelets, there was a direct correlation between ADP-induced biotin- PAC1 binding and binding determined in a conventional 125I-PAC1 binding assay (r = .99; P less than .001). Furthermore, as few as 0.8% activated platelets could be detected by flow cytometry when activated platelets were mixed with unstimulated platelets. In whole blood, unstimulated platelets demonstrated no PAC1- or S12-specific fluorescence, indicating that they did not bind these antibodies. On stimulation with agonists, however, the platelets demonstrated a dose- dependent increase in fluorescence similar to that observed for platelets in plasma or buffer. Low concentrations of ADP and epinephrine, which induce fibrinogen receptors but little secretion, stimulated near-maximal PAC1 binding but little S12 binding. On the other hand, a concentration of phorbol myristate acetate (TPA) that evokes full platelet aggregation and secretion induced maximal PAC1 and S12 binding. Activated platelets could also be analyzed in whole blood samples that had been fixed with paraformaldehyde. These studies demonstrate that activated platelets can be reliably detected in whole blood using activation-dependent monoclonal antibodies and flow cytometry. This technique may be useful to assess the degree of platelet activation and the efficacy of antiplatelet therapy in clinical disorders.
46
Shattil, SJ, M. Cunningham, and JA Hoxie. "Detection of activated platelets in whole blood using activation- dependent monoclonal antibodies and flow cytometry." Blood 70, no. 1 (July 1, 1987): 307–15. http://dx.doi.org/10.1182/blood.v70.1.307.bloodjournal701307.
Full textAbstract:
Platelets may become activated in a number of clinical disorders and participate in thrombus formation. We developed a direct test for activated platelets in whole blood using flow cytometry. Whole blood was incubated with either biotin-PAC1, a monoclonal antibody specific for the fibrinogen receptor on activated platelets, or biotin-S12, an antibody specific for an alpha-granule membrane protein that associates with the platelet surface during secretion. Platelet-bound antibodies were detected with streptavidin conjugated with fluorescein isothiocyanate (FITC) or phycoerythrin (PE). Platelets were differentiated from the larger erythrocytes and WBCs by their light- scatter profile. Alternatively, platelets could be identified with FITC- AP1, an antibody specific for platelet membrane glycoprotein Ib, and analyzed further for PAC1 or S12 binding with PE-streptavidin. No centrifugation or washing steps were required. With gel-filtered platelets, there was a direct correlation between ADP-induced biotin- PAC1 binding and binding determined in a conventional 125I-PAC1 binding assay (r = .99; P less than .001). Furthermore, as few as 0.8% activated platelets could be detected by flow cytometry when activated platelets were mixed with unstimulated platelets. In whole blood, unstimulated platelets demonstrated no PAC1- or S12-specific fluorescence, indicating that they did not bind these antibodies. On stimulation with agonists, however, the platelets demonstrated a dose- dependent increase in fluorescence similar to that observed for platelets in plasma or buffer. Low concentrations of ADP and epinephrine, which induce fibrinogen receptors but little secretion, stimulated near-maximal PAC1 binding but little S12 binding. On the other hand, a concentration of phorbol myristate acetate (TPA) that evokes full platelet aggregation and secretion induced maximal PAC1 and S12 binding. Activated platelets could also be analyzed in whole blood samples that had been fixed with paraformaldehyde. These studies demonstrate that activated platelets can be reliably detected in whole blood using activation-dependent monoclonal antibodies and flow cytometry. This technique may be useful to assess the degree of platelet activation and the efficacy of antiplatelet therapy in clinical disorders.
47
Lötscher, M., H. Amstutz, C. H. Heusser, and K. Blaser. "Specific VDJ gene combinations contribute to the specificity of memory antibodies to the phosphorylcholine hapten." Journal of Immunology 148, no. 11 (June 1, 1992): 3631–35. http://dx.doi.org/10.4049/jimmunol.148.11.3631.
Full textAbstract:
Abstract Based on their fine specificity, two groups of antibodies against the phosphorylcholine (PC) hapten have been described. Group I antibodies react predominantly with the PC moiety of the hapten and group II are directed against the entire hapten including the azophenyl spacer to the protein carrier. We have analyzed the VH gene segment utilization of hybridomas from the memory response to PC by Southern blot analysis and nucleotide sequencing of the functional VDJ rearrangements. Three main specificities of anti-PC antibodies could be distinguished. Anti-PC hybridomas with group I fine specificity utilize the VH1-DFL 16.1-JH1 rearrangement. A major portion of group II antibodies recognized the phenyl-PC part and expressed the same VH1 gene in combination with a member of the SP2 family and JH1 or JH2. The other anti-PC antibodies either used the PJ14-DFL16-JH3 rearrangement in combination with a lambda 1 L chain or a member of the VGam3.8 VH family rearranged to the DFL16.1 and the JH3 gene segments. The PJ14 and VGam3.8 V gene expressing antibodies were directed to the phenyl group and were either not or barely inhibitable by PC chloride. Thus, specific VDJ gene combinations contribute to the fine specificity of antibodies in the memory response to the PC hapten. The use of the S107, Q52, and VGam3.8. VH gene families, together with FL16.1 or SP2 D segments and JH1, JH2, or JH3 results in different fine specificities to the PC, phenyl-PC, or the azophenyl moiety of the PC hapten. These fine specificities of the memory response use V, D, and J segments of the initial T15Id+ response in combination with gene segments usually related to phenyl specificity.
48
Boggs, J. M., G. A. Hashim, E. D. Day, and M. A. Moscarello. "Lipid-induced recognition of a conformational determinant (residues 65 to 83) in myelin basic protein." Journal of Immunology 135, no. 4 (October 1, 1985): 2617–22. http://dx.doi.org/10.4049/jimmunol.135.4.2617.
Full textAbstract:
Abstract The precipitation by antibodies to intact myelin basic protein (BP) and to synthetic peptides containing a sequence based on the region 65 to 83 of bovine BP, S82, S81, S79, and S24, of intact BP in solution or bound to lipid vesicles was compared, using 125I-BP or 14C-DPPC-labeled lipid-BP vesicles. The antipeptide antibodies were shown earlier to recognize conformational determinants which are not expressed in the intact protein in solution. Several anti-BP antibodies precipitated more of the BP free in solution than when bound to lipid vesicles, suggesting that some of the determinants recognized by these antibodies were either sequestered in the bilayer or were altered in conformation. In contrast, one anti-peptide antisera, which had a high titer for the conformational determinant in two of these peptides, S82 and S81, precipitated the protein to a significant degree when it was bound to PG vesicles, even though it did not react with the intact protein in solution. These results indicated that PG was able to confer on the protein the unique peptide conformation recognized by this antibody. PS was less effective, and other lipids were ineffective at conferring this conformation on the protein, supporting earlier results which showed that the conformation of the protein is influenced by the lipid composition of its environment. None of the other anti-peptide antibodies studied bound to the protein either in solution or in lipid vesicles. These results indicate that the lipid environment can sequester or alter the conformation of some antigenic determinants, preventing recognition by some anti-BP antibodies, and can expose or generate other conformational determinants, allowing recognition by an anti-peptide antiserum.
49
Wang, Xuesong, Yu Yan, Tianyu Gan, Xi Yang, Dapeng Li, Dongming Zhou, Qiang Sun, Zhong Huang, and Jin Zhong. "A trivalent HCV vaccine elicits broad and synergistic polyclonal antibody response in mice and rhesus monkey." Gut 68, no. 1 (November 27, 2017): 140–49. http://dx.doi.org/10.1136/gutjnl-2017-314870.
Full textAbstract:
ObjectiveDespite the development of highly effective direct-acting antivirals, a prophylactic vaccine is needed for eradicating HCV. A major hurdle of HCV vaccine development is to induce immunity against HCV with high genome diversity. We previously demonstrated that a soluble E2 (sE2) expressed from insect cells induces broadly neutralising antibodies (NAbs) and prevents HCV infection. The objective of this study is to develop a multivalent HCV vaccine to increase the antigenic coverage.DesignWe designed a trivalent vaccine containing sE2 from genotype 1a, 1b and 3a. Mice and rhesus macaques were immunised with monovalent or trivalent sE2 vaccine, and sera or purified immunoglobulin were assessed for neutralisation against a panel of cell culture-derived virion (HCVcc) of genotype 1–7 in cell culture. Splenocytes from the vaccinated macaques were assessed for HCV-specific T cell response.ResultsWe showed that the trivalent vaccine elicited pangenotypic NAbs in mice, which neutralised HCVcc of all the seven genotypes more potently than the monovalent vaccine. Further analyses demonstrated that each sE2 component of this trivalent vaccine elicited unique spectrum of NAbs which acted synergistically to inhibit HCV infection. Finally, the trivalent vaccine triggered stronger and more uniform multigenotypic neutralising antibody response than the monovalent vaccine in rhesus macaques.ConclusionsIn summary, we developed a trivalent HCV vaccine that induces broad and synergistic-acting neutralising antibodies in mice and non-human primates.
50
Mitchell, Leslie Ann. "Derivation of Sirococcusstrobilinus specific monoclonal antibodies." Canadian Journal of Forest Research 16, no. 5 (October 1, 1986): 939–44. http://dx.doi.org/10.1139/x86-166.
Full textAbstract:
Cloned hydridoma cell lines producing monoclonal antibodies to Sirococcusstrobilinus (a seed-borne fungus causing shoot blight in pine, spruce, and fir seedlings) have been prepared by immunizing BALB/c mice with whole mycelium and fusing their spleen cells with SP2/0-Ag14 myeloma cells. Specificity for S. strobilinus was ascertained by testing the monoclonal antibodies in enzyme-linked immunosorbent assays and by indirect surface immunofluorescence tests against a panel of seven nonrelated commensal fungi obtained from the same seed lot as the Sirococcus isolate used in the derivation of the monoclonal antibodies. The monoclonal antibodies produced by these selected hydridoma cell lines also recognize antigens from other S. strobilinus isolates from diverse host species and tissues. The monoclonal reagents will thus be useful as diagnostic probes for locating S. strobilinus in or on seed and other plant tissues. Preliminary characterization of the morphologic distribution of antigens recognized by the monoclonal antibodies is discussed.