Journal articles on the topic 'CD4'
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Tjønnfjord, G. E., O. P. Veiby, R. Steen, and T. Egeland. "T lymphocyte differentiation in vitro from adult human prethymic CD34+ bone marrow cells." Journal of Experimental Medicine 177, no. 6 (June 1, 1993): 1531–39. http://dx.doi.org/10.1084/jem.177.6.1531.
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Pluripotent lymphohematopoietic stem cells are probably confined to bone marrow cells expressing CD34 surface molecules. To investigate the capacity of adult human CD34+ bone marrow cells to differentiate along the T lymphoid lineage, we plated purified CD34+ cells from healthy adults in liquid culture on adherent thymic stromal cells prepared from HLA- or blood group-mismatched postnatal thymic tissue. We show that purified CD34+CD3-CD4-CD8- bone marrow cells contained progenitors with the ability to differentiate into CD4+ and CD8+ T lymphocytes expressing surface (s)CD3 and T cell receptor alpha/beta in vitro. These progenitors were found in the CD34+CD2+sCD3-CD4-CD8-, CD34+CD7+sCD3-CD4-CD8-, and CD34+CD2+CD7+sCD3-CD4-CD8-, as well as in the CD34+CD2-sCD3-CD4-CD8-, CD34+CD7-sCD3-CD4-CD8-, and CD34+CD2-CD7-sCD3-CD4-CD8- subsets, indicating that T lymphocyte progenitors sensitive to signals mediated by thymic stroma in vitro are not restricted to CD34+ cells already coexpressing early T lymphocyte-associated markers. Finally, we show that T lymphopoiesis was enhanced by c-kit ligand.
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Galy, A., S. Verma, A. Bárcena, and H. Spits. "Precursors of CD3+CD4+CD8+ cells in the human thymus are defined by expression of CD34. Delineation of early events in human thymic development." Journal of Experimental Medicine 178, no. 2 (August 1, 1993): 391–401. http://dx.doi.org/10.1084/jem.178.2.391.
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Studies of the most immature T cell progenitors in the human thymus have been hampered by the lack of markers and assays that define these cells. In this report we used a novel human fetal thymic organ culture system to determine the potential of T cell precursors isolated from human postnatal thymus, to differentiate into CD3+ thymocytes, and to investigate early stages of human T cell development. It was found that thymocytes that lack the markers CD3, CD4, and CD8 (triple negative [TN]) can differentiate in an allogeneic organotypic thymic culture. The capacity of TN thymocytes to differentiate was exclusively confined to the CD34+ population. CD34- TN thymocytes failed to differentiate in this system. In contrast, cloned lines of CD3- thymocytes could only be established from CD34- TN thymocytes. Five subsets of CD3- thymocytes were found with the following phenotype: CD1-TN, CD1+TN, CD1+CD4+CD8-, CD1+CD4+CD8 alpha+ beta-, and CD1+CD4+CD8 alpha beta+. These subpopulations expressed decreasing levels of CD34. The CD1-CD3- population expressed the highest levels of CD34 supporting the notion that this population is the most immature T cell precursor in the thymus, whereas the CD1+CD4+CD8 alpha+ beta+ which did not express CD34 seems to be the most mature of these CD3- populations. This notion is supported by the observations that CD34+ cells isolated from fetal liver, which differentiated into T cells in a FTOC, developed into CD3+ cells via CD1- and CD4+CD8- intermediates. Based on these data, we present a model of early stages in human intrathymic development.
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Federmann, Birgit, Matthias Haegele, Christoph Faul, Wichard Vogel, Lothar Kanz, and Wolfgang A. Bethge. "Immune Reconstitution after Haploidentical Hematopoietic Cell Transplantation: Impact of Reduced Intensity Conditioning and CD3/CD19-Depleted Grafts." Blood 112, no. 11 (November 16, 2008): 1175. http://dx.doi.org/10.1182/blood.v112.11.1175.1175.
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Abstract Haploidentical hematopoietic cell transplantation (HHCT) using CD3/CD19 depleted grafts may lead to faster engraftment and immune reconstitution since grafts contain also graft-facilitating-cells, CD34− progenitors, NK cells, and dendritic cells. Reduced intensity conditioning may also have a positive impact on immune reconstitution following HHCT. 26 adults received CD3/CD19 depleted HHCT after RIC (150–200 mg/m2 fludarabine, 10mg/kg thiothepa, 120 mg/m2 melphalan and 5mg/day OKT-3 (day −5 to +14)) at our institution between 2005–2008. We prospectively evaluated engraftment and immune reconstitution. B-, NK-, T- and T-cell subsets (CD3/8, CD4/8, CD4/45RA/RO), TCR-Vβ repertoire and NK-cell receptors (NKP30, NKP44, NKP46, NKG2D, CD158a/b/e, CD85j, NKG2A, CD161) were analyzed by FACS. Grafts contained 8.8×106 CD34+ (range, 4.3–18.0 ×106), 2.9×104 CD3+ (range, 1.2–9.2×104) and 3.6×107 CD56+ (range, 0.02–23.0 ×107) cells/kg. Engraftment was rapid with a median time to >500 granulocytes/μl of 11 days (range, 9–15) and a median time to >20 000 platelets/μl of 11 days (range, 8–23). Full chimerism was reached on day 14 (median; range, 6–26). NK-cell engraftment was rapid, reaching normal values on day 20 (median of 247 CD16+CD56+CD3− cells/μl (range, 1–886)) with NK cells comprising up to 70% of lymphocytes. B-cell reconstitution was delayed with 81 (range, 0–280) and 335 (range, 11–452) CD19+20+ cells/μl on days 150 and 400, respectively. T-cell reconstitution was impaired with 49 (range, 0–586) and 364 (range, 35–536) CD3+ cells/μl on day 60 and day 150, respectively. We observed an increase of CD3+CD8+ cells in contrast to CD3+CD4+ cells early after HHCT with a median of 24 (range, 0–399) vs 16 (range, 0–257) and 159 (range, 1–402) vs 96 (range, 18–289) cells/μl on day 50 and day 200, respectively. CD4+CD45RA+ T cells increased slowly while CD4+CD45RO+ T cells reconstituted faster with a median of 61 CD4+CD45RO+ cells/μl (range, 0–310) vs 24 CD4+CD45RA+ (range, 0 to 152) on day 100. Within the CD4+CD25+ regulatory T cells there was a slow regeneration with median of 14 CD4+CD25+ cells/μl (range, 0–96) on day 100 and 28 CD4+CD25+ cells/μl (range, 19–160) on day 200. CD14+CD45+ monocytes did not reach normal values within the time of observation with 7 CD14+CD45+ cells/μl (range, 0–21) on day 120 and 7 CD14+CD45+ cells (range, 2–381) on day 400. TCR-Vβ repertoire and NK-cell receptor reconstitution was analyzed so far in 7 and 8 patients, respectively. We found a skewed T-cell repertoire with oligoclonal T-cell expansions to day 100 and normalization after day 200. An increased natural cytotoxicity receptor (NKP30, NKP44, NKP46) and NKG2A, but decreased NKG2D and KIR-expression was observed on NK-cells until day 100. In conclusion, T- and B-cell reconstitution is delayed after HHCT using CD3/CD19 depleted grafts and RIC. However, T-cell reconstitution is faster compared to data published with CD34 selected grafts and myeloablative conditioning. A fast NK-cell reconstitution early after HHCT was observed. Thus a combination of reduced intensity conditioning with CD3/CD19 depleted grafts appears to accelerate the immune recovery after haploidentical stem cell transplantation.
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Márquez, C., C. Trigueros, E. Fernández, and M. L. Toribio. "The development of T and non-T cell lineages from CD34+ human thymic precursors can be traced by the differential expression of CD44." Journal of Experimental Medicine 181, no. 2 (February 1, 1995): 475–83. http://dx.doi.org/10.1084/jem.181.2.475.
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In addition to T-lineage cells, a small proportion of hematopoietic non-T cells are present in the human postnatal thymus. However, the origin of this minor non-T cell thymic compartment is presently unknown. In this study we have analyzed the developmental potential of the earliest human intrathymic precursors, characterized as CD34+ cells expressing intermediate levels of CD44. We show that these CD34+CD44int thymocytes cultured with interleukin 7 were able to develop simultaneously into both T- and non-T (monocytes and dendritic cells) -lineage cells. Both developmental pathways progress through a CD1+CD4+ intermediate stage, currently believed to be the immediate precursor of double positive thymocytes. However, separate progenitors for either T or non-T cells could be characterized within CD1+CD4+ thymocytes by their opposite expression of CD44. Downregulated levels of CD44 identified CD1+CD4+ T-lineage precursors, whereas CD44 upregulation occurred on CD1+CD4+ intermediates that later differentiated into non-T cells. Therefore, commitment of human early intrathymic precursors to either T or non-T cell lineages can be traced by the differential expression of the CD44 receptor.
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Dobrynina, M. A., A. V. Zurochka, M. V. Komelkova, Sh Luo, V. A. Zurochka, Hu Desheng, L. V. Ryabova, and A. P. Sarapultsev. "Studies of CD45+ and CD46+ expression on the peripheral blood lymphocyte subsets of the post-COVID patients." Russian Journal of Immunology 25, no. 4 (October 7, 2022): 431–36. http://dx.doi.org/10.46235/1028-7221-1160-soc.
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The SARS-CoV-2 virus can enter the cells using S1 viral spike (S) protein, not only by binding to ACE2, but also through other cellular receptors. These candidate receptors include CD46, which, like CD45, belongs to pan-leukocyte receptors and is expressed on all types of lymphocytes. In turn, SARS-CoV-2 infection is accompanied by damage to almost all compartments of the immune system, mainly T lymphocytes. The purpose of the study was to evaluate the expression levels of CD45+ and CD46+ in various subpopulations of lymphocytes in patients who had undergone SARS-CoV-2 infection.
72 patients who had undergone SARS-CoV-2 infection were examined. Using flow cytometry technique, we determined CD45+ and CD46+ (panleukocyte marker for lymphocyte gating), CD45+ and CD46+, CD3+ (T lymphocytes), CD45+ and CD46+, CD3+, CD4+ (helper inducers), CD45+ and CD46+, CD3+, CD8+ (cytotoxic T-lymphocytes), CD45+ and CD46+, CD3+, CD56+ (TNK cells) CD45+ and CD46+, CD3-, CD56+ (natural killers), CD45+ and CD46+, CD3-, CD19+ (B lymphocytes), CD45+ and CD46+, CD3+, CD4+, CD25+ (activated helpers, early activation of lymphocytes), CD45+ and CD46+, CD3+, HLA-DR (activated T lymphocytes late activation of lymphocytes). Our studies have shown that a decrease in CD46+ expression in T lymphocytes (CD3+) is accompanied by similar decrease of its expression in cytotoxic T lymphocytes (CD3+, CD8+), TNK (CD3+, CD56+), as well as in helpers T carrying markers of early activation (CD3+, CD4+, CD25+). At the same time, the most pronounced decrease was observed both among total T lymphocytes and cytotoxic T cells. In these patients, the expression level of CD46+ in B lymphocytes was slightly increased. Recent data suggest that there is no involvement of CD46 receptor on B lymphocytes. Our data suggest that the SARS-CoV-2 virus may affect the CD46 receptor. Such exposure may lead to promotion of the long-COVID (post-COVID) symptoms in such patients, thus requiring new approaches to correction of these disorders.
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Chatila, T. A., and R. S. Geha. "Phosphorylation of T cell membrane proteins by activators of protein kinase C." Journal of Immunology 140, no. 12 (June 15, 1988): 4308–14. http://dx.doi.org/10.4049/jimmunol.140.12.4308.
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Abstract Activation of the enzyme protein kinase C (PKC) plays an important role in T cell activation. We investigated the phosphorylation of CD2, CD3, CD4, CD5, CD7, CD8, CD28 (Tp44), CD43 (sialophorin, gp115), and LFA-1 after incubation of human PBMC with the (PKC) activator PMA. These proteins were chosen for their role in transmembrane signal transduction (CD2, CD3, CD5, CD28, CD43), cell-cell interaction and adhesion (CD2, CD4, CD8, and LFA-1), or involvement in immunodeficiency states (CD43, CD7). CD5, CD7, CD43, and the alpha-chain of LFA-1 were found to be constitutively phosphorylated. PMA induced rapid hyperphosphorylation of CD5, CD7, and CD43, but not of the LFA-1 alpha-chain, and induced the phosphorylation of CD3, CD4, CD8 and of the LFA-1 beta-chain. PMA did not cause the phosphorylation of CD2 and CD28. PMA-induced phosphorylation was partially inhibited by the PKC inhibitor 1-(5-isoquinolinylsulfonyl)-2-methylpiperazine dihydrochloride. Finally, the T cell activator Con A, which binds to the CD3/TCR complex was shown to induce a profile of protein phosphorylation similar to that observed with PMA. We conclude that PKC-mediated phosphorylation of T cell Ag may represent an important regulatory mechanism that governs the process of T cell activation.
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Landolfi, M. M., N. Van Houten, J. Q. Russell, R. Scollay, J. R. Parnes, and R. C. Budd. "CD2-CD4-CD8- lymph node T lymphocytes in MRL lpr/lpr mice are derived from a CD2+CD4+CD8+ thymic precursor." Journal of Immunology 151, no. 2 (July 15, 1993): 1086–96. http://dx.doi.org/10.4049/jimmunol.151.2.1086.
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Abstract MRL lpr/lpr (lymphoproliferative, lpr) mice demonstrate an age-dependent lymphoproliferation and development of autoimmunity. Characteristic of the lymphoproliferation in these mice is the accumulation of large numbers of CD4-CD8-(CD4-8-),CD3+ T lymphocytes in their lymph nodes. The development of the CD4-8- cells, which also aberrantly express B220 and CD44 (Pgp-1) but are CD2-, has been shown to be thymus dependent. An unusual feature of lpr CD4-8-T lymphocytes is that although they appear unresponsive to stimulation, as defined by proliferation and IL-2 production, they have undergone thymic negative selection. As thymic deletion normally occurs at the CD4+CD8+ (CD4+8+) stage, this raises the dilemma that lpr CD4-8- T lymphocytes have either previously been CD4+8+, or they are able to undergo thymic selection as CD4-8- cells. We have addressed this question by examining the methylation status of the CD8 gene in MRL lpr CD4-8- lymph node cells. Demethylation of the CD8 gene has been shown to be an indicator of previous CD8 expression. We find that the CD8 gene in lpr CD4-8- lymph node cells, as well as in the abnormal B220+ CD4-8- lpr thymocytes, is demethylated, suggesting that these cells have previously expressed CD8. In addition, we find that the lpr CD4+8+ thymocyte population contains an increased percentage of atypical B220+, CD44+ cells that are virtually all CD2+. Taken together, these data are consistent with the lpr CD2-CD4-8- population of LNC having arisen from a CD2+ CD4+8+ thymic stage of differentiation.
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Quinn, Tyler, Jung-Hyun Kim, Amanda Strauch, Tianzhou Wu, Jeffery Powell, Raymond Roberge, Ronald Shaffer, and Aitor Coca. "Physiological Evaluation of Cooling Devices in Conjunction With Personal Protective Ensembles Recommended for Use in West Africa." Disaster Medicine and Public Health Preparedness 11, no. 5 (March 17, 2017): 573–79. http://dx.doi.org/10.1017/dmp.2016.209.
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AbstractObjectiveCooling devices (CDs) worn under personal protective equipment (PPE) can alleviate some of the heat stress faced by health care workers responding to the Ebola outbreak in West Africa.MethodsSix healthy, young individuals were tested while wearing 4 different CDs or no cooling (control) under PPE in an environmental chamber (32°C/92% relative humidity) while walking (3 METs, 2.5 mph, 0% grade) on a treadmill for 60 minutes. Exercise was preceded by a 15-minute stabilization period and a 15-minute donning period.ResultsThe control condition resulted in a significantly higher rectal temperature (Tre) at the end of the exercise than did all CD conditions (CD1,P=0.004; CD2,P=0.01; CD3,P=0.000; CD4,P=0.000) with CD1 and CD2 resulting in a higher Trethan CD3 and CD4 (P<0.05). The control condition resulted in a higher heart rate (HR) at the end of exercise than did the CD3 (P=0.01) and CD4 (P=0.009) conditions, whereas the HR of the CD1 and CD2 conditions was higher than that of the CD3 and CD4 conditions (P<0.05). Weight loss in the control condition was higher than in the CD3 (P=0.003) and CD4 (P=0.01) conditions. Significant differences in subjective measurements of thermal stress were found across conditions and time.ConclusionsUse of CDs can be advantageous in decreasing the negative physiological and subjective responses to the heat stress encountered by health care workers wearing PPE in hot and humid environments. (Disaster Med Public Health Preparedness. 2017;11:573–579)
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Vicente, A., A. Varas, R. S. Acedón, E. Jiminez, J. J. Mulqoz, and A. G. Zapata. "Appearance and Maturation of T-Cell Subsets During Rat Thymus Ontogeny." Developmental Immunology 5, no. 4 (1998): 319–31. http://dx.doi.org/10.1155/1998/24239.
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In previous papers, we have described the ontogenetical development of thymic stromal-cell components (epithelium, macrophages, dendritic cells) of Wistar rats. Here, we correlate those results with the maturation of rat T-cell precursors along the fetal and postnatal life. First T-cell precursors, which colonize the thymus anlage around days 13-14 of gestation, largely express CD45, CD43, CD53, and Thy 1 cell markers, and in a lesser proportion the OX22 antigen. Rat CD3-CD4-CD8-thymocytes present in the earliest stages of gestation could be subdivided in three major cell subpopulations according to the CD44 and CD25 expression: CD44-/+CD25-→ CD44+CD25+→ CD44+CD25-On fetal days 17-18, a certain proportion of CD4-CD8-cells weakly,express the TcRβchain, in correlation with the appearance of the first immature CD4-CD8+thymocytes. This cell subpopulation, in progress to the CD4+CD8+stage, upregulates CD8αbefore the CD8βchain, expresses the CD53 antigen, and exhibits a high proliferative rate. First mature thymocytes arising from the DP (CD4+CD8+) cells appear on fetal days 20-21. Then, the CD4+:CD8+cell ratio is ≤1 changing to adult values (2-3) just after birth. Also, the percentage of VβTcR repertoire covered in adult thymus is reached during the postnatal period, being lower during the fetal life. Finally, in correlation with the beginning of thymocyte emigration to the periphery a new wave of T-cell maturation apparently occurs in the perinatal rat thymus.
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Suda, T., and A. Zlotnik. "Origin, differentiation, and repertoire selection of CD3+CD4-CD8- thymocytes bearing either alpha beta or gamma delta T cell receptors." Journal of Immunology 150, no. 2 (January 15, 1993): 447–55. http://dx.doi.org/10.4049/jimmunol.150.2.447.
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Abstract It has been widely accepted that CD3+CD4-CD8- T cells expressing TCR-alpha beta or TCR-gamma delta (found in the thymus as well as in the periphery) represent lineages distinct from either CD4+CD8- and CD4-CD8+ single-positive T cells expressing TCR-alpha beta. However, the origin, differentiation pathway, and TCR-repertoire selection of CD3+CD4-CD8- T cells remain controversial. We demonstrate that CD3+CD4-CD8- thymocytes can be separated into three subsets based on their expression of heat-stable Ag (HSA) and CD44. Our results further suggest the following: 1) the HSA+ subset represents a pre-selection population, although the HSA- subset is a postselection subset; 2) the high incidence of V beta 8.2 usage among CD3+CD4-CD8- thymocytes is a result of positive selection, rather than a predetermined event at a precursor cell level; 3) the maturation of CD3+CD4-CD8- thymocytes proceeds along the following differentiation pathway: HSA+CD44(-)-->HSA-CD44(-)-->HSA-CD44+. Both TCR-alpha beta +CD4-CD8- and TCR-gamma delta +CD4-CD8- thymocytes show similar differentiation processes; 4) CD3+CD4-CD8-cells directly differentiate from CD25+CD3-CD4-CD8- thymocytes which include precursor cells for both the CD3+CD4-CD8- and the CD4+CD8-/CD4-CD8+ lineages. Taken together, these results suggest that the CD25+CD3-CD4-CD8- stage of thymocyte differentiation represents a branching point for either the CD4+CD8- or CD4-CD8+ single-positive lineages or the CD3+CD4-CD8- lineages.
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Suda, T., and A. Zlotnik. "In vitro induction of CD8 expression on thymic pre-T cells. II. Characterization of CD3-CD4-CD8 alpha + cells generated in vitro by culturing CD25+CD3-CD4-CD8- thymocytes with T cell growth factor-beta and tumor necrosis factor-alpha." Journal of Immunology 149, no. 1 (July 1, 1992): 71–76. http://dx.doi.org/10.4049/jimmunol.149.1.71.
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Abstract We previously reported that CD25 (IL-2R p55)-positive CD3-CD4-CD8- murine thymocytes can be induced to express CD8 alpha (Lyt-2) by transforming growth factor-beta plus TNF-alpha in the presence of IL-7 (which is necessary to maintain the viability and differentiation capacity of CD25+CD3-CD4-CD8- thymocytes in vitro). The majority of cells recovered after 2 to 3 days from these cultures expressed CD8 alpha (but not CD3 or CD4). In this study, we have characterized these in vitro generated CD3-CD4-CD8 alpha + thymocytes and compared them with normal CD3-CD4-CD8+ thymocytes. Unlike normal CD3-CD4-CD8+ thymocytes that express CD8 alpha and CD8 beta (Lyt-3-chain) simultaneously, only a fraction of in vitro generated CD3-CD4-CD8 alpha + cells expressed CD8 beta. However, along with the induction of CD8 alpha and CD8 beta expression, the expression of other T cell differentiation markers (including CD2, CD25, and CD44) also changed in a manner corresponding to physiologic differentiation. Cell-surface phenotyping suggests that CD8 alpha + beta - cells are less mature than CD8 alpha + beta + cells. These in vitro generated CD3-CD4-CD8 alpha + thymocytes expanded and differentiated into the CD4+CD8+ stage as well as mature (CD3+) single positive (CD4+CD8-) and CD4-CD8+) stages in fetal thymus organ culture that had been depleted of lymphoid cells by treatment with 2-deoxyguanosine. The latter observation indicates that these in vitro generated CD3-CD4-CD8 alpha + thymocytes are responsive to other differentiation-inducing signals (including those that induce CD4) that exist in fetal thymus organ culture. These results suggest that in vitro generated CD3-CD4-CD8 alpha + thymocytes represent intermediate differentiation stages between CD25+CD3-CD4-CD8- and CD3-CD4-CD8+ cells found in normal thymus.
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Koulova, L., S. Y. Yang, and B. Dupont. "Identification of the anti-CD3-unresponsive subpopulation of CD4+, CD45RA+ peripheral T lymphocytes." Journal of Immunology 145, no. 7 (October 1, 1990): 2035–43. http://dx.doi.org/10.4049/jimmunol.145.7.2035.
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Abstract The majority of peripheral CD4+ T lymphocytes proliferate in vitro in response to anti-CD3 in presence of autologous APC. The present study describes a subpopulation of CD4+ T cells that cannot be activated and progress into cell cycle by stimulation with anti-CD3 plus APC or with mitogenic combinations of anti-CD2. The in vitro responses of these anti-CD3-unresponsive CD4+ T cells were investigated with a panel of mAb to CD2, CD3, and CD28, and found to be similar to those previously observed for mature thymocytes: only the combination of anti-CD2 plus anti-CD28 produced cell proliferation. Anti-CD3-unresponsive T cells were CD45RA+, but represented only 14 to 22% of the CD4+, CD45RA+ T cell population. Activation with anti-CD2 plus anti-CD28 mAb resulted in major changes in the cell surface phenotype and functional properties: a loss of CD45RA+ occurred and an increased expression of CD45RO, CD29, and CD58 (LFA3), as well as a gain in responsiveness to anti-CD3 and anti-CD2. This change in CD45 phenotype from CD45RA to CD45RO occurs in both the anti-CD3-responsive and in the anti-CD3-unresponsive subsets of the CD45RA+, CD4+ cells after cell proliferation. The anti-CD3-unresponsive subset may represent a pool of not yet fully differentiated peripheral T cells. The acquisition of anti-CD3 responsiveness could occur as a consequence of Ag priming or by an Ag-independent mechanism. Involvement of the CD28 Ag in this process is suggested from the present study.
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Zybleva, S. V., and S. L. Zyblev. "CD3+CD4+CD8+ T-LYMPHOCYTES COUNT IN PROGNOSIS OF EARLY KIDNEY ALLOGRAFT DYSFUNCTION." Journal of the Grodno State Medical University 18, no. 1 (March 2020): 17–20. http://dx.doi.org/10.25298/2221-8785-2020-18-1-17-20.
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Muench, Marcus O., Maria Grazia Roncarolo, and Reiko Namikawa. "Phenotypic and Functional Evidence for the Expression of CD4 by Hematopoietic Stem Cells Isolated From Human Fetal Liver." Blood 89, no. 4 (February 15, 1997): 1364–75. http://dx.doi.org/10.1182/blood.v89.4.1364.
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Abstract Expression of the CD4 antigen was observed on human fetal liver, fetal bone marrow (BM), and umbilical cord blood progenitors expressing high levels of CD34. Using clonal and liquid-culture assays, CD4+ CD34++ Lin− (lineage = CD3, CD8, CD10, CD14, CD15, CD16, CD19, CD20, and glycophorin A) fetal liver progenitors were found to have a greater proliferative potential than CD4− CD34++ Lin− progenitors, whereas the CD4− fraction was more enriched for erythroid progenitors. Both the CD4+ and the CD4− progenitor subpopulations also gave rise to multilineage engraftment upon transplantation into human fetal bone fragments, supportive of B-lymphoid and myeloid growth, or into human fetal thymic fragments, supportive of T-cell growth, implanted in scid/scid (SCID) mice. However, in SCID-hu mice transplanted with graded doses of donor cells ranging from 2.0 × 102 to 2.0 × 104 cells, BM reconstitution by the CD4+ fraction of CD34++ Lin− cells was more frequent than by the CD4− fraction when low numbers of cells were injected. These functional data strongly suggest that stem cells reside among CD4+ CD34++ Lin− fetal liver cells. This hypothesis was further supported by the observations that CD4+ CD34++ Lin− fetal liver cells were enriched for CDw90+ (Thy-1), CD117+ (kit), CD123+, HLA-DR+, CD7−, CD38−, CD45RA−, CD71−, CD115− (fms), and rhodamine 123dull cells, a phenotypic profile believed to represent fetal stem cells. Furthermore, all CD4+ CD34++ Lin− fetal liver cells also expressed CD13 and CD33.
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Зыблева, С. В., and С. Л. Зыблев. "Cluster Analysis of Leukocyte Subpopulations in Kidney Transplantation." Гематология. Трансфузиология. Восточная Европа, no. 2 (November 8, 2021): 168–75. http://dx.doi.org/10.34883/pi.2021.7.2.005.
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Цель. Выявить варианты иммунного реагирования у пациентов при трансплантации почки на основе кластерного анализа, характеризующие течение посттрансплантационного периода. Материалы и методы. Обследовано 104 реципиента почечного трансплантата с терминальной стадией хронической болезни почек, которым выполнена трансплантация аллогенной почки, а также 90 здоровых добровольцев, составивших группу сравнения. Оценены уровни лейкоцитов с использованием метода проточной цитометрии по безотмывочной технологии с использованием моноклональных антител (Beckman Coulter и BD, США) CD4 PС7, CD8 FITC, CD3 PС5.5, CD3 FITC, CD45 PerCP, CD19 APC, CD56+ CD16 PE, CD3 PC5.5, HLA-DR APC, CD38 PE, CD4 FITC, CD3 PC5.5, CD8 APC, CD69 PE, CD127 PЕ, CD25 АРС CD154 PЕ, CD3 APCAF750, IgD FITC, CD27 PC5.5, CD5 APC-AF750, CD40РЕ, CD86 РЕ, CD14 PС7, CD64 FITC, CD86 PЕ LIN PE, CD123 PC7, Anti-HLADR APC-AF750 в объемах, рекомендуемых фирмой-производителем. Результаты и обсуждение. В результате проведенного исследования разработана система оценки иммунного статуса реципиента почечного трансплантата, обеспечивающая персонифицированный мониторинг, анализ и прогнозирование течения посттрансплантационного периода. Описаны виды регуляторных клеточных сетей и их синергический потенциал при трансплантации почки. В основе толерогенного иммунологического комплекса у пациентов после трансплантации почки лежат межклеточные взаимодействия, имеющие иерархическую систему, основа которой представлена кооперацией клеток CD3+CD4+CD25+highCD127+low, CD3+CD4-CD8-, CD3+CD4+CD69+, CD3+CD16+CD56+, CD19+CD5+ и LIN-HLA-DR+CD11c-CD123+. Воснове гиперергического иммунологического комплекса при почечной аллотрансплантации лежат избыточно активированные компоненты иммунного ответа: CD3+CD8+CD69+, CD3+CD4+CD8+, CD3+CD8+CD38+, CD19+CD86+, CD19+IgD+CD27-, CD3-CD16+CD56+, CD3+CD38+, CD14+lowCD86+, и LIN-HLA-DR+CD11c+CD123клетки. Выводы. Выделенные иммунофенотипы позволят осуществить персонифицированный подход к диагностике и лечению пациентов с различными вариантами иммунного реагирования при трансплантации почки. При выявлении иммунофенотипа, соответствующего толерогенному варианту иммунного ответа, терапию пациента в отдаленном посттрансплантационном периоде возможно проводить с учетом низкого иммунологического риска и минимизацией иммуносупрессивной нагрузки. Purpose. To identify the variants of immune response in patients with kidney transplantation on the base of cluster analysis that characterize the course of the post-transplant period. Materials and methods. We examined 104 kidney transplant recipients with end-stage chronic kidney disease, who underwent allograft transplantation, as well as 90 healthy volunteers, who made up the comparison group. Their leukocyte levels were assessed using no-wash flow cytometry with monoclonal antibodies (Beckman Coulter and BD, USA) CD4 PС7, CD8 FITC, CD3 PС5.5, CD3 FITC, CD45 PerCP, CD19 APC, CD56+ CD16 PE, CD3 PC5.5, HLA-DR APC, CD38 PE, CD4 FITC, CD3 PC5.5, CD8 APC, CD69 PE, CD127 PЕ, CD25 АРС CD154 PЕ, CD3 APC-AF750, IgD FITC, CD27 PC5.5, CD5 APC-AF750, CD40РЕ, CD86 РЕ, CD14 PС7, CD64 FITC, CD86 PЕ LIN PE, CD123 PC7, Anti-HLADR APC-AF750 in the volumes recommended by the manufacturer. Results and discussion. As a result of the conducted study, the system for assessing the immune status of a kidney transplant recipient was developed, which provides personalized monitoring, analysis, and prediction of the course of the post-transplant period. The types of regulatory cellular networks and their synergistic potential in kidney transplantation are described. The tolerogenic immunological complex in patients after kidney transplantation is based on intercellular interactions that have a hierarchical system, the base of which is represented by the cooperation of CD3+CD4+CD25+highCD127+low, CD3+CD4-CD8-, CD3+CD4+CD69+, CD3+CD16+CD56+, CD19+CD5+ and LIN-HLA-DR+CD11c-CD123+ cells. The hyperergic immunological complex in renal allotransplantation is based on over-activated components of the immune response: CD3+CD8+CD69+, CD3+CD4+CD8+, CD3+CD8+CD38+, CD19+CD86+, CD19+IgD+CD27-, CD3-CD16+CD56+, CD3+CD38+, CD14+lowCD86+, and LIN-HLA-DR+CD11c+CD123- cells. Conclusion. The detected immunotypes will let to implement the personalized approach to the diagnostics and treatment of patients with various types of immune response in kidney transplantation. If an immunophenotype corresponding to a tolerogenic variant of the immune response is identified, the patient’s therapy in the long-term post-transplant period can be carried out taking into account the low immunological risk and minimizing the immunosuppressive load.
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Blue, M. L., J. F. Daley, H. Levine, K. R. Branton, and S. F. Schlossman. "Regulation of CD4 and CD8 surface expression on human thymocyte subpopulations by triggering through CD2 and the CD3-T cell receptor." Journal of Immunology 142, no. 2 (January 15, 1989): 374–80. http://dx.doi.org/10.4049/jimmunol.142.2.374.
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Abstract Human thymocytes bearing the CD4 and/or CD8 antigens can be fractionated into cells with an immature and more mature phenotype based on their quantitative expression of the CD3 Ag (J. Immunol. 138:3108; J. Immunol. 139:1065). We show that the expression of CD4 and CD8 on thymocyte subpopulations with low CD3 (CD3L) and high CD3 (CD3H) is regulated by activation through the CD2 molecule and perturbation of the CD3-T cell receptor complex (CD3-Ti). Similar to its previously reported effects on peripheral T cells, PMA was able to induce the down-regulation of surface CD4, but not CD8, on thymocyte subpopulations. PMA could induce CD4 and CD8 phosphorylation in both CD3L and CD3H fractions. These results suggest that if changes in phosphorylation represent the mechanism by which CD4 and CD8 are able to transmit signals, this mechanism is operative in both CD3L and CD3H subpopulations. Treatment with anti-T11(2) and anti-T11(3) antibodies (CD2 activation pathway) resulted in partial down-regulation of CD4 but not CD8 surface expression on both CD3L and CD3H thymocytes. Similar treatment had no detectable effect on peripheral T cells. The down-regulation of surface CD4 induced by activation via CD2 could be inhibited by treatment of thymocytes with anti-CD3 antibodies. Treatment of thymocytes with anti-CD3 alone or following CD2 activation induced the selective down-regulation of surface CD8 within 15 minutes. These results suggest that CD2 and CD3-Ti triggering may regulate CD4 and CD8 surface expression on thymocytes. Furthermore, these results suggest that "cross-talk" between the CD2 and CD3-Ti pathway of activation may involve CD4 and CD8 molecules.
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Chen, H., and W. E. Paul. "Cultured NK1.1+ CD4+ T cells produce large amounts of IL-4 and IFN-gamma upon activation by anti-CD3 or CD1." Journal of Immunology 159, no. 5 (September 1, 1997): 2240–49. http://dx.doi.org/10.4049/jimmunol.159.5.2240.
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Abstract NK1.1+ CD4+ T cells produce more IL-4 and IFN-gamma in response to stimulation with anti-CD3 than conventional CD4+ T cells that have been primed to be, respectively, Th2 or Th1 cells. Furthermore, NK1.1+ CD4+ T cells produce IL-4 even if cultured in the absence of IL-4, whereas conventional CD4+ T cells require IL-4 to develop into IL-4 producers. The IFN-gamma-producing capacity of NK1.1+ CD4+ T cells is enhanced by IL-12. In addition, NK1.1+ CD4+ T cells produce substantial amounts of IL-3, IL-5, and IL-10 upon anti-CD3 stimulation. NK1.1+ CD4+ T cells can be stimulated to produce IL-4 by culture with L cells expressing CD1 (L-CD1), but primary IL-4 production was rather slow and weak. Restimulation of L-CD1-activated NK1.1+ CD4+ T cells with L-CD1 gave rise to a much stronger and more rapid response, yielding IL-4 production comparable with anti-CD3 activation of cells initially primed with anti-CD3. L-CD1 stimulation of L-CD1-primed cells resulted in far less IFN-gamma than that elicited by anti-CD3 from cells that had been primed with anti-CD3, but such production is substantially increased by adding IL-12 to the culture. The differing patterns of cytokine production by NK1.1+ CD4+ T cells suggest that these cells may have complex effects on the priming of conventional T cells and may not simply drive such cells to the acquisition of a Th2 phenotype.
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Agarwal, Ashima, and Stephen I. Fisher. "Angioimmunoblastic T- Cell Lymphoma by Flow Cytometric Analysis." Blood 106, no. 11 (November 16, 2005): 4707. http://dx.doi.org/10.1182/blood.v106.11.4707.4707.
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Abstract Background: Angioimmunoblastic T cell lymphoma (AILT) is a peripheral T- cell lymphoma characterized by systemic disease, a polymorphous infiltrate involving lymph nodes with prominent proliferation of high endothelial venules and follicular dendritic cells. We present a novel technique for the presumptive diagnosis of AILT cases by flow cytometric analysis. The diagnosis of AILT was confirmed by histolopathologic features, immunohistochemical stains and molecular studies. Method: Specimens: Fresh groin excision lymph nodes. Flow cytometric analysis utilizing side scatter versus CD45. Case 1 (figure 1): The gated population consists of 10–12% B cells and 56–61% T cells. CD3, CD4 and CD5 stain 56–61% of the gated cells, while CD7 stains 41% of the gated cells, consistent with aberrant partial loss of CD7 by T cells. A population of abnormal T cells expresses CD10 with CD2, CD3, CD4 and CD7 but not with CD19 (B cells) consistent with aberrant CD10 coexpression by a subset of the T cells (12–19% of the gated cells). Staining for CD34 highlights less than 2% immature cells staining while CD64 stains less than 2% monocytic cells. Surface kappa and lambda staining does not show restriction (kappa: lambda 1–2:1). Cytoplasmic kappa and lambda staining is not interpretable due to non-specific antibody binding. Case 2 (figure 2): The gated population consists of 69% T- cells and 28–33% B- cells. A population of abnormal T cells expresses CD10 with CD2, CD3, CD4 and CD7 but not CD19 consistent with aberrant CD10 expression by a subpopulation of T cells (8–11% of gated cells). Intracellular B cell kappa lambda light chain restriction is not present and there is no aberrant co-expression of CD5, CD10 or CD43 by B cells. The diagnosis of AILT was confirmed by standard morphologic, immunoperoxidase and molecular methods subsequently in both of these cases. Discussion: Aberrant CD10 expression on neoplastic T cells has been shown with immunohistochemical staining of paraffin embedded tissue in AILT. Our cases show aberrant coexpression of CD10 by CD2, CD3, CD4 and CD7 positive T cells in more than 10% of gated cells by Flow cytometric analysis. Conclusion: Flow Cytometric analysis is a useful and reproducible tool for immunophenotyping AILT cells and should be considered for presumptive diagnosis of AILT. Figure Figure Figure Figure
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Terstappen, LW, S. Huang, and LJ Picker. "Flow cytometric assessment of human T-cell differentiation in thymus and bone marrow." Blood 79, no. 3 (February 1, 1992): 666–77. http://dx.doi.org/10.1182/blood.v79.3.666.bloodjournal793666.
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Using multidimensional flow cytometry we have defined and quantified the human T-cell differentiation pathway, focusing on those events occurring among the most immature thymocytes and putative bone marrow (BM) T-precursors. Early thymocytes were found to express the CD34 antigen and consisted of a mean 1.2% of cells within human pediatric (n = 9) and 2.0% in fetal thymi (n = 4). All CD34+ thymocytes were atypical blast by morphology, expressed intracytoplasmatic, but not cell surface, CD3, and were cell surface CD2+, CD5+, CD7+, CD38+, CD45+, CD45RA+, CD49d+, and LECAM-1(Leu8)high. CD34high thymocytes lacked surface expression of CD4 and CD8, but as CD34 expression diminished there was a coordinate increase in CD4 levels, followed by the appearance of CD8. The expression of CD1 and CD10 also increased concomitant with the loss of CD34, whereas expression of LECAM-1 diminished with CD34 downregulation. The differential expression of these antigens on early thymocytes (as well as the number of thymocytes displaying these patterns) was highly reproducible among the nine pediatric and four fetal specimens examined, suggesting a precise, stereotyped regulation of early differentiation events. Cell populations with antigen expression patterns suggestive of pluripotent stem cell (CD34high, CD38-), or non-T-lineage committed stem cells (CD34+, CD33+ or CD34+, CD19+) were not identified in either fetal or pediatric thymi (sensitivity = 1/10(4)). The presence of cells with the antigenic profile of the earliest CD34+ thymocytes was explored in human BM. Putative BM T-cell precursors with the appropriate phenotype (CD34+, CD7+, CD5+, CD2+, LECAM-1high) were readily identified in fetal specimens (constituting +/- 2% of the CD34+ population), but could not be reliably detected in adults. In contrast with thymi, only 13% of these cells expressed cytoplasmatic CD3, suggesting the presence of the immediate precursor of the putative prothymocyte population. This was further supported by the detection of CD34bright, CD7+, CD2-, CD5-, LECAM-1moderate cells in fetal specimens. Our results document the flow of cell surface differentiation during T-lymphopoiesis and suggest that T-lineage features are first acquired in the BM. The ability to reproducibly identify and isolate T-cell precursor populations of precisely defined maturational stage in marrow and thymus by multiparameter flow cytometry will facilitate characterization of the molecular events controlling T-lineage differentiation.
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Li, Yanchun, Shucheng Hua, Hang Gao, Wenxue Li, and Yaping Zhu. "Changes of cell surface markers CD3, CD4, CD44, CD62L on mouse spleen lymphocytes after culture in vitro (HYP7P.303)." Journal of Immunology 192, no. 1_Supplement (May 1, 2014): 119.18. http://dx.doi.org/10.4049/jimmunol.192.supp.119.18.
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Abstract Object To Study the changes of cell surface markers CD3, CD4, CD44 and CD62L on mouse spleen lymphocytes after culture. Method Spleen cells were separated from Balb/c mice and asthmatic mice and lymphocytes were isolated by using lymphocyte separation liquid. We cultured lymphocytes with RPMI 1640 medium(10% FBS) in cell incubator for 3 days. Then the four cell surface markers were detected by using flow cytometry. Results In normal mice, 19.09% of lymphocytes stained positively for CD3 and CD4, among these cells 98.61% was CD44+ T cell and 68.71% was CD62L+ T cell before the culture. After culture with ConA for 3 days, CD3+CD4+ and CD3+CD4+CD44+ T cell population was down regulated to 8.96% and 71.82%, respectively. While CD62L almost disappeared, accounting for 11.27% only. The similar results were obtained even cells cultured without ConA present. For asthmatic mice, CD3+CD4+ T cell accounted for 20.33% in total lymphocytes, among which 97.72% was CD44+ and 75.74% was CD62L+ before the culture. After culture, CD3+CD4+T cell accounted for 10.4%, among which CD44+ only 55.5% and CD62L+ only 2.63%. The similar results were obtained when cells were cultured with OVA. Conclusion In both normal and asthmatic mice, CD3,CD4,CD44 and CD62L on spleen lymphocytes were down regulated after culture in vitro regardless with or without stimulating factors, especially CD62L, almost disappeared. This should be given full consideration when these markers are detected after cell culture.
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Ledbetter, J. A., G. L. Schieven, F. M. Uckun, and J. B. Imboden. "CD45 cross-linking regulates phospholipase C activation and tyrosine phosphorylation of specific substrates in CD3/Ti-stimulated T cells." Journal of Immunology 146, no. 5 (March 1, 1991): 1577–83. http://dx.doi.org/10.4049/jimmunol.146.5.1577.
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Abstract In lymphocytes, CD45 regulates the increase in cytoplasmic calcium concentration that occurs after receptor cross-linking. Here we show that T cell receptor complex (CD3/Ti)-mediated inositol phosphate production was inhibited by CD45 ligation in Jurkat cells. CD3/Ti signaling in normal T cells was also inhibited by CD45 ligation, but coupling of CD4 with CD3/Ti gave augmented calcium signals that were entirely resistant to the inhibitory effect of CD45. In contrast, CD3-induced T cell proliferation was suppressed by immobilized CD45 mAb even in the presence of CD4 mAb. The effect of CD45 and CD4 ligation on tyrosine phosphorylation during T cell activation was directly examined by immunoblotting with anti-phosphotyrosine. Using immobilized mAb, CD45 ligation suppressed the tyrosine phosphorylation of specific substrates induced by CD3/Ti stimulation, including almost complete suppression of 150-, 36-, and 35-kDa proteins and partial suppression of 76- and 80-kDa proteins. Other tyrosine-phosphorylated proteins induced by CD3/Ti stimulation, including 135- and 21-kDa proteins, were not suppressed by simultaneous ligation of CD3/Ti and CD45. Simultaneous ligation of CD3 and CD4 enhanced tyrosine phosphorylation of all substrates, but did not overcome the CD45-mediated suppression of tyrosine phosphorylation of the 35- and 36-kDa proteins. The CD45-mediated suppression of phospholipase C activation is therefore modulated by association with CD4 without altering the specific inhibition of tyrosine phosphorylation and T cell proliferation after co-ligation of CD45 and CD3/Ti.
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Saizawa, M. K., E. Hug, S. Haque, P. Portoles, S. Suzuki, and K. Eichmann. "Autoreactivity of low but not of high CD4 variants of an antigen-specific, I-A-restricted mouse T cell clone." Journal of Immunology 148, no. 3 (February 1, 1992): 702–9. http://dx.doi.org/10.4049/jimmunol.148.3.702.
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Abstract Variant lines expressing high and low surface densities of the accessory molecule CD4 have been developed by repeated preparative flow cytometric cell sortings from the murine Th cell clone D10.G4.1 (D10). The high CD4 variant line (D10H) fully maintained the original I-Ak restricted specificity for conalbumin of wild-type D10 cells. In contrast, the low CD4 variant line (D10L) showed a strong autoreactivity to I-Ak carrying stimulator cells alone which was only slightly augmented by addition of conalbumin. Cell surface molecules other than CD4, including TCR, CD3, CD11a, CD2, CD45, CD44, and MHC class I, remained identical on D10H and D10L sublines as on D10 wild-type cells. The possibility that D10L cells had suffered alterations of their TCR-alpha beta was excluded by demonstrating their reactivity with a panel of eight different anti-clonotypic mAb specific for various epitopes of the D10 TCR. By limiting dilution analysis we show that the majority of responding cells of D10L sublines were autoreactive. Although the reactivity for allogeneic I-A also increased as compared with D10H cells, a clear preference for self-I-Ak was maintained so that a true autoreactive phenotype was evident. The results indicate that the surface concentration of CD4 has a decisive influence on self-non-self discrimination of MHC class II-restricted Th cells.
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Giorgio, Selma, Marcia Regina M. Santos, Anita H. Straus, Helio K. Takahashi, and Clara Lúcia Barbiéri. "Effect of Glycosphingolipids Purified from Leishmania (Leishmania) amazonensis Amastigotes on Human Peripheral Lymphocytes." Clinical Diagnostic Laboratory Immunology 10, no. 3 (May 2003): 469–72. http://dx.doi.org/10.1128/cdli.10.3.469-472.2003.
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ABSTRACT The effect of purified glycosphingolipids from Leishmania (Leishmania) amazonensis on human lymphoproliferation, on expression of human lymphocyte and monocyte markers (CD3, CD4, CD8, CD14, CD19, and CD45), and on lymphocyte protein kinase C activity was analyzed.
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Yang, S. Y., S. Rhee, K. Welte, and B. Dupont. "Differential in vitro activation of CD8-CD4+ and CD4-CD8+ T lymphocytes by combinations of anti-CD2 and anti-CD3 antibodies." Journal of Immunology 140, no. 7 (April 1, 1988): 2115–20. http://dx.doi.org/10.4049/jimmunol.140.7.2115.
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Abstract Purified peripheral blood T lymphocytes and the CD8-CD4+ and CD4-CD8+ T cell subsets, exhaustively depleted of APC have been studied for their capacity to respond to mAb directed against the CD3-Ti Ag-specific TCR complex and against the CD2 SRBCR. It is demonstrated that high affinity IL-2R can be readily induced by either anti-CD3 and/or anti-CD2 stimulation. However, IL-2 production can be observed only in the CD4+CD8- T cell subset. These results clearly contrast data obtained with purified CD4-CD8+ T cells, which are able to proliferate when the CD3-Ti complex is activated in the presence of APC. The data presented in the present study demonstrate that a simplified model for T cell activation and clonal expansion of the two major T cell subsets involve only the CD3-Ti complex and the CD2 Ag. Under conditions where the activation signals for the T cells are restricted only to the activation of CD3-Ti and CD2, the CD4+CD8- T cells respond with IL-2 production and expression of high affinity IL-2R, whereas the CD4-CD8+ T cell subset depends on exogenous IL-2 provided by the CD4+CD8- cells. These data do not, however, exclude an involvement of other cell-surface signals for regulation and control of T cell activation and T cell effector functions.
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Geppert, T. D., and P. E. Lipsky. "Association of various T cell-surface molecules with the cytoskeleton. Effect of cross-linking and activation." Journal of Immunology 146, no. 10 (May 15, 1991): 3298–305. http://dx.doi.org/10.4049/jimmunol.146.10.3298.
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Abstract The association of various surface molecules with the cytoskeleton in resting peripheral blood T cells was examined by assaying the capacity of detergent to solubilize them. Cytoskeletal association was assessed by staining T cells with a fluorescein-conjugated mAb, resuspending the cells in buffer with or without the nonionic detergent, NP-40, and determining the capacity of the detergent to remove the mAb from the cell surface by using flow microfluorimetry. MAb to CD3, the TCR, and CD45 were completely removed from the cell surface by detergent. In contrast, 7 to 50% of mAb to CD2, CD4, CD8, CD11a/CD18, CD44, and class I MHC molecules were resistant to detergent solubilization, demonstrating that a fraction of these molecules was constitutively associated with the cytoskeleton. The effect of cross-linking these molecules with a mAb and a secondary goat anti-mouse Ig was also examined. Cross-linking CD3 or the TCR induced cytoskeletal association of these molecules. In addition, cross-linking increased the fraction of CD2, CD4, CD8, CD11a/CD18, CD44, and class I MHC molecules that was associated with the cytoskeleton. In contrast, cross-linking CD45 did not induce an association with the cytoskeleton. The effect of T cell activation on the cytoskeletal association of these molecules was also examined. Stimulation of T cells with ionomycin and PMA greatly increased the expression of CD2 and CD44 without increasing the number of molecules associated with the cytoskeleton. Stimulation with PMA alone had no effect on the expression of CD2 or CD44, but was found to decrease the percentage of these molecules associated with the cytoskeleton. Stimulation with ionomycin and PMA increased both the expression of class I MHC molecules and the number of molecules associated with the cytoskeleton proportionally. Finally, stimulation with ionomycin and PMA decreased CD3 expression, but increased the number of CD3 molecules associated with the cytoskeleton. The data establish a pattern of cytoskeletal association of T cell-surface molecules that is a characteristic of each individual molecule and can be altered by cross-linking. Moreover, the results indicate that the association of various T cell surface molecules with the cytoskeleton is a dynamic process that varies with the state of activation and or differentiation of the cells.
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Matsuyama, T., A. Yamada, K. Deusch, J. Sleasman, J. F. Daley, Y. Torimoto, and T. Abe. "Cytochalasins enhance the proliferation of CD4 cells through the CD3-Ti antigen receptor complex or the CD2 molecule through an effect on early events of activation." Journal of Immunology 146, no. 11 (June 1, 1991): 3736–41. http://dx.doi.org/10.4049/jimmunol.146.11.3736.
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Abstract Cytochalasins are known to inhibit or enhance the proliferation of T cells induced by mitogens in a concentration-dependent fashion. To clarify the mechanism by which cytochalasins enhance T cell proliferation, we examined which activation pathways and events in signal transduction were affected by cytochalasins. We also examined subsets of CD4 cells for a preferential response to cytochalasins. Cytochalasins enhanced the proliferation of CD4 cells induced by optimal doses of anti-CD3 antibody or suboptimal doses of anti-CD2 antibodies. Cytochalasins, at low concentrations, enhanced the rise in intracellular Ca2+ and production of IP3 in CD4 cells activated by anti-CD2 or CD3 antibodies. Cytochalasins also enhanced the modulation of CD3 induced by anti-CD3 antibody. These results suggest that cytochalasins enhance the proliferation of CD4 cells by affecting early events in signal transduction after activation through the CD3-Ti Ag-receptor complex or CD2 molecule. At the doses used, cytochalasins appear to interact with cytochalasin-binding sites in the cell membrane. Cytochalasins predominantly enhanced CD3-mediated proliferation in the CD29-subset of CD4 cells.
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Mittler, R. S., B. M. Rankin, and P. A. Kiener. "Physical associations between CD45 and CD4 or CD8 occur as late activation events in antigen receptor-stimulated human T cells." Journal of Immunology 147, no. 10 (November 15, 1991): 3434–40. http://dx.doi.org/10.4049/jimmunol.147.10.3434.
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Abstract Activation of human PBL T cells with solid phase anti-CD3 mAb or during the course of an MLR response gives rise to the association of CD4 or CD8 molecules with the protein tyrosine phosphatase, CD45, on the cell surface. This paired association of cell-surface molecules occurs late in the activation cycle and appears to be dependent upon Ti-CD3-mediated signaling because mitogen-driven activation does not induce formation of the complex. Maximal association occurred 72 to 96 h after exposure to anti-CD3 mAb on both CD4+ and CD8+ T cells. In contrast, association between CD8 and CD45 during an MLR response did not occur until day 6 of a MLR whereas CD4-CD45 association was detected by 72 h of culture. The kinetics of association between CD4 or CD8 and CD45 was measured by fluorescence resonance energy transfer and confirmed by immunoprecipitation of dithiobis succinimidylpropionate or disuccinimidyl suberate cross-linked 125I-labeled resting or activated T cells. The molecules that co-precipitated with either CD4 or CD8 and had an apparent kDa of 180 to 205 could be immunodepleted with anti-CD45 mAb. Furthermore, CD4 or CD8 immunoprecipitates from 96-h activated T cells contained significant levels of protein tyrosine phosphatase activity whereas corresponding immunoprecipitates from resting or recently activated T cells showed little protein tyrosine phosphatase activity. This association may allow CD45 to engage and dephosphorylate lck or another CD4- or CD8-associated substrate in order to reset the receptor complex to receive a new set of stimuli. Our observations suggest that synergistic signaling provided as a consequence of CD4 or CD8 association with the TCR after antigenic stimulation may develop on a different temporal scale than that observed after soluble anti-CD4+ anti-CD3 heteroconjugate antibody cross-linking.
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Rettig, Michael P., Julie K. Ritchey, Bruno Nervi, Mark L. Bonyhadi, and John F. DiPersio. "Comparison of the Division Rate and Proliferative Capacity of Naive and Ex Vivo Activated T Cells after Allogeneic Bone Marrow Transplantation." Blood 104, no. 11 (November 16, 2004): 2133. http://dx.doi.org/10.1182/blood.v104.11.2133.2133.
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Abstract Maintaining T cell function after ex vivo manipulation remains a major challenge in adoptive immunotherapy. We previously showed that murine T cells activated ex vivo with anti-CD3 and anti-CD28 antibody-coated magnetic beads (CD3/CD28 beads) retain greater GVHD-inducing potential than cells activated with either soluble anti-CD3 antibody alone or plate bound anti-CD3 and anti-CD28 antibodies after allogeneic BMT. However, CD3/CD28 bead activated T cells still exhibit reduced GVHD-inducing potential compared to naïve T cells. In this study, we used CFSE and congenic mice to monitor the proliferative kinetics of naïve and CD3/CD28 bead activated, transduced, and selected T cells in the same mouse after allogeneic and syngeneic BMT. High efficiency (>50%) gene transfer of our chimeric suicide gene, CD34/TK, into C57BL/6 (B6) murine T cells was achieved 24 h after CD3/CD28 bead activation and gene-modified cells were purified to >98% by CD34 immunomagnetic selection 48 h later. CD34/TK+ T cells were then rested for 3 days in medium containing 10 U/mL IL-2. Purified CD34/TK+ (CD45.1+) and naïve (CD45.2+) T cells from B6 mice were then labeled with CFSE, mixed 1/1 (3e6 total T cells), and injected, along with T cell depleted B6 (CD45.1+) BM, into lethally irradiated allogeneic (BALB/c, CD45.2+) or syngeneic (B6, CD45.2+) recipients. Mice were sacrificed daily up to 6 days after BMT to assess donor T cell engraftment, division, and phenotype by five color flow cytometry. The CD34/TK+ donor T cells (both CD4+ and CD8+ T cells) underwent 1–2 rounds of division within 24 h after infusion. In contrast, <5% of the naïve T cells divided during the first 24 h after infusion. Thereafter, the CD34/TK+ and naïve CD4+ and CD8+ T cells exhibited similar division kinetics between days 1 and 4 after BMT. At 3 days after BMT, both CD34/TK+ and naïve CD4+ and CD8+ T cells were detected in the 8 cell divisions discernible by CFSE, with approximately equal percentages (5%–15%) of cells in each division cycle. However, virtually all of the CD34/TK+ and naïve CD4+ and CD8+ T cells had divided more than 7 times by day 4 after allogeneic BMT. In contrast, <10% of the CD34/TK+ or naïve CD4+ and CD8+ T cells had undergone more than 7 cell divisions in the syngeneic recipients. Interestingly, the CD34/TK+ T cells exhibited a dramatic decrease in expansion compared with the naïve T cells between days 4 and 6 after allogeneic BMT. Although ~ equal percentages of CD34/TK+ and naïve T cells were observed 4 days after infusion, >6-fold and 10-fold more naïve CD4+ and CD8+ T cells, respectively, were detected in the allogeneic recipients at day 6 after BMT. This effect was specific to the allogeneic response, because we observed no difference in expansion between the CD34/TK+ and naïve CD4+ and CD8+ T cells in the syngeneic recipients. Phenotypically, both the CD34/TK+ and naïve CD4+ and CD8+ cells upregulated CD25 expression after 4 divisions, upregulated CD69 and CD44 expression after 1 to 2 divisions, and downregulated CD62L expression. In summary, CD3/CD28 bead activated, transduced, and selected T cells exhibit decreased expansion compared to naïve T cells after injection into allogeneic recipients. Ongoing studies are evaluating whether this decrease in expansion is caused by activation induced cell death, altered trafficking, or a decrease in the proliferative capacity of the ex vivo manipulated cells.
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Blue, M. L., D. A. Hafler, K. A. Craig, H. Levine, and S. F. Schlossman. "Phosphorylation of CD4 and CD8 molecules following T cell triggering." Journal of Immunology 139, no. 12 (December 15, 1987): 3949–54. http://dx.doi.org/10.4049/jimmunol.139.12.3949.
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Abstract CD4 and CD8 molecules have been implicated in the regulation of T cell activation. In the present study, CD4 and CD8 were modified by increased phosphorylation when T cell clones or T cells were either exposed to phorbol-12-myristate- 13-acetate or were triggered via the CD3-T cell receptor complex. Activation of T cells through the CD2 sheep erythrocyte binding protein, using anti-T11(2) and -T11(3) antibodies, also resulted in CD4 and CD8 phosphorylation. These findings suggest that signals derived from two different receptor pathways can converge and result in similar molecular modifications of CD4 and CD8. Furthermore, phorbol myristate acetate treatment or activation via the CD2 pathway induced phosphorylation of the CD4 and CD8 molecules of thymocytes, suggesting that these molecules may be functional in thymus. Together, our findings indicate that CD4 and CD8 phosphorylation is a consequence of T cell triggering, and suggest that CD4 and CD8 phosphorylation may represent a molecular signaling mechanism among the CD3-T cell receptor complex, CD2, CD4, and CD8.
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Kim, Dongku, Reina E. Mebius, John D. MacMicking, Steffen Jung, Tom Cupedo, Yaneth Castellanos, Jaerang Rho, et al. "Regulation of Peripheral Lymph Node Genesis by the Tumor Necrosis Factor Family Member Trance." Journal of Experimental Medicine 192, no. 10 (November 20, 2000): 1467–78. http://dx.doi.org/10.1084/jem.192.10.1467.
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Proper lymph node (LN) development requires tumor necrosis factor–related activation-induced cytokine (TRANCE) expression. Here we demonstrate that the defective LN development in TRANCE−/− mice correlates with a significant reduction in lymphotoxin (LT)αβ+α4β7+CD45+CD4+CD3− cells and their failure to form clusters in rudimentary mesenteric LNs. Transgenic TRANCE overexpression in TRANCE−/− mice results in selective restoration of this cell population into clusters, and results in full LN development. Transgenic TRANCE-mediated restoration of LN development requires LTαβ expression on CD45+ CD4+CD3− cells, as LNs could not be induced in LTα−/− mice. LTα−/− mice also showed defects in the fate of CD45+CD4+CD3− cells similar to TRANCE−/− mice. Thus, we propose that both TRANCE and LTαβ regulate the colonization and cluster formation by CD45+ CD4+CD3− cells in developing LNs, the degree of which appears to correlate with the state of LN organogenesis.
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Terstappen, LW, S. Huang, and LJ Picker. "Flow cytometric assessment of human T-cell differentiation in thymus and bone marrow." Blood 79, no. 3 (February 1, 1992): 666–77. http://dx.doi.org/10.1182/blood.v79.3.666.666.
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Abstract Using multidimensional flow cytometry we have defined and quantified the human T-cell differentiation pathway, focusing on those events occurring among the most immature thymocytes and putative bone marrow (BM) T-precursors. Early thymocytes were found to express the CD34 antigen and consisted of a mean 1.2% of cells within human pediatric (n = 9) and 2.0% in fetal thymi (n = 4). All CD34+ thymocytes were atypical blast by morphology, expressed intracytoplasmatic, but not cell surface, CD3, and were cell surface CD2+, CD5+, CD7+, CD38+, CD45+, CD45RA+, CD49d+, and LECAM-1(Leu8)high. CD34high thymocytes lacked surface expression of CD4 and CD8, but as CD34 expression diminished there was a coordinate increase in CD4 levels, followed by the appearance of CD8. The expression of CD1 and CD10 also increased concomitant with the loss of CD34, whereas expression of LECAM-1 diminished with CD34 downregulation. The differential expression of these antigens on early thymocytes (as well as the number of thymocytes displaying these patterns) was highly reproducible among the nine pediatric and four fetal specimens examined, suggesting a precise, stereotyped regulation of early differentiation events. Cell populations with antigen expression patterns suggestive of pluripotent stem cell (CD34high, CD38-), or non-T-lineage committed stem cells (CD34+, CD33+ or CD34+, CD19+) were not identified in either fetal or pediatric thymi (sensitivity = 1/10(4)). The presence of cells with the antigenic profile of the earliest CD34+ thymocytes was explored in human BM. Putative BM T-cell precursors with the appropriate phenotype (CD34+, CD7+, CD5+, CD2+, LECAM-1high) were readily identified in fetal specimens (constituting +/- 2% of the CD34+ population), but could not be reliably detected in adults. In contrast with thymi, only 13% of these cells expressed cytoplasmatic CD3, suggesting the presence of the immediate precursor of the putative prothymocyte population. This was further supported by the detection of CD34bright, CD7+, CD2-, CD5-, LECAM-1moderate cells in fetal specimens. Our results document the flow of cell surface differentiation during T-lymphopoiesis and suggest that T-lineage features are first acquired in the BM. The ability to reproducibly identify and isolate T-cell precursor populations of precisely defined maturational stage in marrow and thymus by multiparameter flow cytometry will facilitate characterization of the molecular events controlling T-lineage differentiation.
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He, You-Wen, and Michael J. Bevan. "High Level Expression of Cd43 Inhibits T Cell Receptor/CD3-Mediated Apoptosis." Journal of Experimental Medicine 190, no. 12 (December 20, 1999): 1903–8. http://dx.doi.org/10.1084/jem.190.12.1903.
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In a screen designed to identify genes that regulate T cell receptor (TCR)/CD3-mediated apoptosis, we found that high level expression of CD43 protected T cell hybridomas from activation-induced cell death. The protection appears to result from its capacity to block Fas-mediated death signals rather than from inhibition of the upregulation of Fas and/or Fas ligand after T cell stimulation. We found that peripheral CD4+ T cells can be divided into two subsets based on the level of CD43 surface expression. The CD4+CD43low subset exhibits a naive T cell phenotype, being CD62LhighCD45RBhighCD44low, whereas CD4+CD43high cells exhibit a memory phenotype, being CD62LlowCD45RBlowCD44high. Recent studies have demonstrated that engagement of TCR and Fas induces naive CD4+ T cells to undergo apoptosis, and the same treatment enhances the proliferation of memory CD4+ T cells. We confirm here that peripheral CD4+CD43high T cells are resistant to TCR/CD3-mediated cell death. These results suggest that the expression levels of CD43 on naive and memory CD4+ T cells determine their susceptibility to Fas-dependent cell death and that high level expression of CD43 may be used as a marker to define CD4+ memory T cells. Expression of CD43 provides a novel mechanism by which tumor cells expressing abnormally high levels of CD43 may escape Fas-mediated killing.
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Ruggiero, G., E. Martinez Cáceres, A. Voordouw, E. Noteboom, D. Graf, R. A. Kroczek, and H. Spits. "CD40 expressed on thymic epithelial cells provides costimulation for proliferation but not for apoptosis of human thymocytes." Journal of Immunology 156, no. 10 (May 15, 1996): 3737–46. http://dx.doi.org/10.4049/jimmunol.156.10.3737.
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Abstract Human thymic epithelial cells express CD40, so we examined the possible role of CD40 in activation of thymocytes. We observed that both CD4+CD8- and CD4-CD8+ thymocytes proliferate after stimulation by anti-CD3 mAb in the presence of cultured thymic epithelial cells. Costimulation of CD4+ thymocytes by thymic epithelial cells is partly inhibited by an anti-CD40 mAb, but this mAb has no effect on costimulation of CD8+ thymocytes. The selective costimulatory ability of CD40 for CD4+ thymocytes was confirmed in experiments in which thymocytes were stimulated with anti-CD3 in the presence of murine P815 cells transfected with CD40 cDNA. The level of costimulation induced by P815-CD40 was comparable with that induced by P815 cells expressing CD80 (B7.1). Treatment of thymocytes with the Ca2+ ionophore ionomycin and the phorbol ester PMA or with anti-CD3 mAb resulted in up-regulation of the CD40 ligand, suggesting that this molecule is involved in CD40-mediated costimulation of human thymocytes. Costimulation of thymocytes by CD80 strongly increased anti-CD3-induced death of fetal thymocytes. In contrast, costimulation by CD40 did not increase anti-CD3-mediated apoptosis of these thymocytes. To confirm that CD40 does not affect anti-CD3-induced cell death, we established a variant of the Jurkat T leukemic cell line that constitutively expresses CD40L and analyzed the sensitivity of this cell line for activation-induced apoptosis. In contrast to CD80, CD40 failed to increase anti-CD3-mediated apoptosis in CD40L+ Jurkat cells, whereas both CD40 and CD80 strongly increased IL-2 production induced by anti-CD3. These findings suggest that costimulation by CD40 is involved in clonal expansion of CD4+ thymocytes but not in activation-induced cell death.
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Hanna, Zaher, Elena Priceputu, Pavel Chrobak, Chunyan Hu, Véronique Dugas, Mathieu Goupil, Miriam Marquis, Louis de Repentigny, and Paul Jolicoeur. "Selective Expression of Human Immunodeficiency Virus Nef in Specific Immune Cell Populations of Transgenic Mice Is Associated with Distinct AIDS-Like Phenotypes." Journal of Virology 83, no. 19 (July 15, 2009): 9743–58. http://dx.doi.org/10.1128/jvi.00125-09.
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ABSTRACT We previously reported that CD4C/human immunodeficiency virus (HIV)Nef transgenic (Tg) mice, expressing Nef in CD4+ T cells and cells of the macrophage/dendritic cell (DC) lineage, develop a severe AIDS-like disease, characterized by depletion of CD4+ T cells, as well as lung, heart, and kidney diseases. In order to determine the contribution of distinct populations of hematopoietic cells to the development of this AIDS-like disease, five additional Tg strains expressing Nef through restricted cell-specific regulatory elements were generated. These Tg strains express Nef in CD4+ T cells, DCs, and macrophages (CD4E/HIVNef); in CD4+ T cells and DCs (mCD4/HIVNef and CD4F/HIVNef); in macrophages and DCs (CD68/HIVNef); or mainly in DCs (CD11c/HIVNef). None of these Tg strains developed significant lung and kidney diseases, suggesting the existence of as-yet-unidentified Nef-expressing cell subset(s) that are responsible for inducing organ disease in CD4C/HIVNef Tg mice. Mice from all five strains developed persistent oral carriage of Candida albicans, suggesting an impaired immune function. Only strains expressing Nef in CD4+ T cells showed CD4+ T-cell depletion, activation, and apoptosis. These results demonstrate that expression of Nef in CD4+ T cells is the primary determinant of their depletion. Therefore, the pattern of Nef expression in specific cell population(s) largely determines the nature of the resulting pathological changes.
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Chirmule, N., T. W. McCloskey, R. Hu, V. S. Kalyanaraman, and S. Pahwa. "HIV gp120 inhibits T cell activation by interfering with expression of costimulatory molecules CD40 ligand and CD80 (B71)." Journal of Immunology 155, no. 2 (July 15, 1995): 917–24. http://dx.doi.org/10.4049/jimmunol.155.2.917.
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Abstract One mechanism of the immune suppression in HIV infection has been postulated as being caused by the interaction of HIV envelope glycoprotein gp120 with CD4 molecules. Thus, pretreatment of purified peripheral blood T cells or CD4+ T cell clones with gp120 (or an anti-CD4 mAb) results in inhibition of anti-CD3 mAb-induced proliferative responses. In this study, we have analyzed the role of the interacting pairs of costimulatory molecules, CD28-B71 (CD80) and CD40 ligand (CD40L)-CD40, to elucidate further the mechanism of HIV gp120-induced inhibitory effects on T cell functions. Interactions between CD28-B71 and CD40L-CD40 were found to be essential for the anti-CD3 mAb-induced T cell proliferation, as demonstrated by up-regulation of B71 and CD40L and the ability of anti-B71 and anti-CD40L mAbs to inhibit this response. Pretreatment of CD4+ T cells with gp120 before CD3 ligation with anti-CD3 mAb resulted in failure of up-regulation of CD40L on T cells and B71 on APC. Exogenous addition of anti-CD28 mAb overcame the inhibitory effect of gp120 on anti-CD3 mAb-induced T cell proliferation. We conclude that binding of gp120 to CD4 molecules on T cells may interrupt the sequential cascade of intercellular interaction involving 1) Ag/MHC class II-TCR/CD4, 2) CD40L-CD40, and 3) B71-CD28. These studies indicate that the CD4-gp120 interaction results in dysregulation of expression of costimulatory molecules, CD40L, and B71 expression on T cells and APC, respectively, thereby contributing to the T cell hyporesponsiveness in HIV infection.
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Dolzhanskiy, A., RS Basch, and S. Karpatkin. "Development of human megakaryocytes: I. Hematopoietic progenitors (CD34+ bone marrow cells) are enriched with megakaryocytes expressing CD4." Blood 87, no. 4 (February 15, 1996): 1353–60. http://dx.doi.org/10.1182/blood.v87.4.1353.bloodjournal8741353.
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CD34 is expressed by essentially all human hematopoietic progenitors including cells of the megakaryocyte (MK) lineage. We have previously reported CD4 expression by some human MK (Blood 81:2,664, 1993). To study the role of maturation on CD4 expression by MK, we examined CD34+ bone marrow cells for their expression of CD41 (GPIIb-GPIIIa) and CD4 with specific monoclonal antibody (MoAb)-fluorochrome conjugates and for DNA polyploidization with propidium iodide or 7-aminoactinomycin D (7-AAD). Surprisingly, MK were at least 20-fold more common in the CD34+ progenitor pool (approximately 10%) than in the more mature CD34+ population (approximately 0.5%) of low density bone marrow cells. CD4 expression correlated with markers of immaturity in that CD4 was enriched among CD34+ cells, and the proportion of CD4+ MK declined with increasing ploidy. Almost all CD34+ polyploid ( > or = 8N) cells were CD4+. Despite these correlations with immaturity, CD34+CD4+ MK precursors were unable to produce MK colony-forming units (CFU-MK) when cultured under conditions that supported the growth of CFU-MK from CD34+CD4- MK lineage cells. MK became polyploid before the loss of either CD34 or CD4 expression. The presence of CD4 on these cells correlates with the onset of endomitotic reduplication and is associated with the loss of the ability of these cells to undergo normal mitotic division. The role of CD4 on immature MK as a differentiation antigen and/or receptor for the human immunodeficiency virus (HIV)-1 virus remains to be determined.
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Ledbetter, JA, GL Schieven, VM Kuebelbeck, and FM Uckun. "Accessory receptors regulate coupling of the T-cell receptor complex to tyrosine kinase activation and mobilization of cytoplasmic calcium in T- lineage acute lymphoblastic leukemia." Blood 77, no. 6 (March 15, 1991): 1271–82. http://dx.doi.org/10.1182/blood.v77.6.1271.1271.
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Abstract T-lineage acute lymphoblastic leukemia (T-ALL) cells have abundant cytoplasmic CD3/Ti but express low amounts on the cell surface and are deficient in CD3/Ti-mediated signal transduction. Nevertheless, plating T-ALL cells on dishes containing immobilized anti-CD3 monoclonal antibodies with a source of growth factors induced the expression of CD25 (interleukin-2 receptor alpha chain) and stimulated the formation of blast colonies in 12 of 14 cases studied. The proliferative response to CD3 ligation was modulated by the presence of antibodies to the CD2, CD4, or CD8 accessory T-cell receptors. The effect of these accessory receptors on signal transduction mediated by CD3/Ti was next investigated by monitoring cytoplasmic calcium concentration [( Ca2+]i) and by measuring tyrosine phosphorylation after stimulation. Crosslinking CD3, CD2, CD4, or CD8 alone did not induce cytoplasmic calcium mobilization in T-ALLs, but crosslinking the accessory receptors with CD3/Ti induced calcium responses in three of the T-ALLs and enhanced calcium responses in three of the T-ALL cell lines, including HPB-ALL, MOLT-4, and CEM. Crosslinking CD4 but not CD2 with CD3/Ti greatly enhanced tyrosine phosphorylation of multiple substrates in comparison with crosslinking either CD4 or CD3/Ti separately on both normal mature T cells and the CEM T-ALL cell line. Thus, CD4 regulates CD3/Ti signal transduction in T-ALL cells through the tyrosine phosphorylation of substrates whereas CD2 may regulate [Ca2+]i signal transduction through a separate mechanism.
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Ledbetter, JA, GL Schieven, VM Kuebelbeck, and FM Uckun. "Accessory receptors regulate coupling of the T-cell receptor complex to tyrosine kinase activation and mobilization of cytoplasmic calcium in T- lineage acute lymphoblastic leukemia." Blood 77, no. 6 (March 15, 1991): 1271–82. http://dx.doi.org/10.1182/blood.v77.6.1271.bloodjournal7761271.
Full textAbstract:
T-lineage acute lymphoblastic leukemia (T-ALL) cells have abundant cytoplasmic CD3/Ti but express low amounts on the cell surface and are deficient in CD3/Ti-mediated signal transduction. Nevertheless, plating T-ALL cells on dishes containing immobilized anti-CD3 monoclonal antibodies with a source of growth factors induced the expression of CD25 (interleukin-2 receptor alpha chain) and stimulated the formation of blast colonies in 12 of 14 cases studied. The proliferative response to CD3 ligation was modulated by the presence of antibodies to the CD2, CD4, or CD8 accessory T-cell receptors. The effect of these accessory receptors on signal transduction mediated by CD3/Ti was next investigated by monitoring cytoplasmic calcium concentration [( Ca2+]i) and by measuring tyrosine phosphorylation after stimulation. Crosslinking CD3, CD2, CD4, or CD8 alone did not induce cytoplasmic calcium mobilization in T-ALLs, but crosslinking the accessory receptors with CD3/Ti induced calcium responses in three of the T-ALLs and enhanced calcium responses in three of the T-ALL cell lines, including HPB-ALL, MOLT-4, and CEM. Crosslinking CD4 but not CD2 with CD3/Ti greatly enhanced tyrosine phosphorylation of multiple substrates in comparison with crosslinking either CD4 or CD3/Ti separately on both normal mature T cells and the CEM T-ALL cell line. Thus, CD4 regulates CD3/Ti signal transduction in T-ALL cells through the tyrosine phosphorylation of substrates whereas CD2 may regulate [Ca2+]i signal transduction through a separate mechanism.
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Budd, R. C., J. Q. Russell, N. van Houten, S. M. Cooper, H. Yagita, and J. Wolfe. "CD2 expression correlates with proliferative capacity of alpha beta + or gamma delta + CD4-CD8- T cells in lpr mice." Journal of Immunology 148, no. 4 (February 15, 1992): 1055–64. http://dx.doi.org/10.4049/jimmunol.148.4.1055.
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Abstract The T lymphocytes that accumulate in vast numbers in the lymphoid tissues of lpr/lpr (lpr) mice express a TCR-alpha beta that is polyclonally rearranged, and yet is devoid of surface CD4 or CD8 (CD4-8-) as well as CD2. lpr CD2- alpha beta + CD4-8- T cells exhibit an apparent block in signal transduction, in that when activated they produce little or no IL-2 and proliferate minimally in the absence of exogenous IL-2. In contrast to the predominant hyporesponsive alpha beta + CD4-8- T cells, we observe that a minor subset (1 to 2%) of lpr lymph node CD4-8- cells expresses a TCR-gamma delta and can proliferate upon activation with PMA and ionomycin in the absence of exogenous IL-2. Furthermore, these responsive gamma delta T cells express surface CD2. The functional and phenotypic distinctions of lpr gamma delta T cells led us to identify an analogous minor (4 to 10%) subset of alpha beta + CD4-8- cells in lpr thymus and lymph nodes that does express CD2. Similar to the gamma delta subset, these CD2+ alpha beta + CD4-8- cells are also capable of proliferation and IL-2 production. Thus the capacity for IL-2 production and proliferation by a small proportion of lpr CD4-8- T cells, either alpha beta + or gamma delta +, correlates with their expression of surface CD2. This correlation is supported by the observation that the lpr liver contains actively cycling alpha beta + CD4-8- lymphocytes that are strikingly enriched for CD2 expression. Consequently, unlike the vast proportion of abnormal lpr CD2- CD3+ CD4-8- cells, the CD2+ CD3+ CD4-8- T cells may not express the basic lpr defect, or else are not affected by its presence. These studies suggest that expression of the lpr abnormality may be restricted to a particular T cell lineage. This functional correlation with CD2 expression may be more broadly applicable to phenotypically similar subsets of normal thymocytes, and possibly peripheral tolerized T lymphocytes.
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Gordeychuk, I. V., A. I. Tukhvatulin, S. P. Petkov, M. A. Abakumov, S. A. Gulyaev, N. M. Tukhvatulina, T. V. Gulyaeva, M. I. Mikhaylov, D. Y. Logunov, and M. G. Isaguliants. "Assessment of the Parameters of Adaptive Cell-Mediated Immunity in Naïve Common Marmosets (Callithrix jacchus)." Acta Naturae 10, no. 4 (December 15, 2018): 63–69. http://dx.doi.org/10.32607/20758251-2018-10-4-63-69.
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Common marmosets are small New World primates that have been increasingly used in biomedical research. This report presents efficient protocols for assessment of the parameters of adaptive cell-mediated immunity in common marmosets, including the major subpopulations of lymphocytes and main markers of T- and B-cell maturation and activation using flow cytometry with a multicolor panel of fluorescently labelled antibodies. Blood samples from eight common marmosets were stained with fluorescently labeled monoclonal antibodies against their population markers (CD45, CD3, CD20, CD4, CD8) and lymphocyte maturation and activation markers (CD69, CD62L, CD45RO, CD107a and CD27) and analyzed by flow cytometry. Within the CD45+ population, 22.75.5% cells were CD3- CD20+ and 67.66.3% were CD3+CD20-. The CD3+ subpopulation included 55.75.5% CD3+CD4+CD8- and 34.33.7% CD3+CD4-CD8+ cells. Activation and maturation markers were expressed in the following lymphocyte proportions: CD62L on 54.010.7% of CD3+CD4+ cells and 74.412.1% of CD3+CD8+ cells; CD69 on 2.71.2% of CD3+CD4+ cells and 1.20.5% of CD3+CD8+ cells; CD45RO on 1.60.6% of CD3+CD4+ cells and 1.80.7% of CD3+CD8+ cells; CD107a on 0.70.5% of CD3+CD4+ cells and 0.50.3% of CD3+CD8+ cells; CD27 on 94.62.1% of CD3+ cells and 8.93.9% CD20+ cells. Female and male subjects differed in the percentage of CD3+CD4+CD45RO+ cells (1.90.5 in females vs 1.10.2 in males; p 0.05). The percentage of CD20+CD27+ cells was found to highly correlate with animals age (r = 0.923, p 0.005). The basal parameters of adaptive cell-mediated immunity in nave healthy marmosets without markers of systemic immune activation were obtained. These parameters and the described procedures are crucial in documenting the changes induced in common marmosets by prophylactic and therapeutic immune interventions.
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Deusch, K., J. F. Daley, H. Levine, A. J. Languet, P. Anderson, S. F. Schlossman, and M. L. Blue. "Differential regulation of Ca2+ mobilization in human thymocytes by coaggregation of surface molecules." Journal of Immunology 144, no. 8 (April 15, 1990): 2851–58. http://dx.doi.org/10.4049/jimmunol.144.8.2851.
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Abstract Variations in intracellular Ca2+ levels in developing thymocytes are likely to play a major role in both the activation-associated differentiation of thymocytes and in the selection or clonal deletion of cells. Here we examine the role of CD4, CD8, CD2, and CD45 in the regulation of intracellular Ca2+ levels in mature and immature thymocytes. Mature and immature thymocytes, distinguished on the basis of their CD5 expression, were analyzed simultaneously for their ability to mobilize Ca2+ after coaggregation of their CD3/TCR with other thymic surface Ag. Flow cytometric analysis by using Indo-1 showed that coaggregation of CD4, CD8, and CD2 with CD3/TCR clearly enhances a minimal signal delivered via CD3/TCR on immature thymocytes. Coaggregation with class I MHC had no discernible effect. The responsiveness of immature thymocytes correlated strictly with CD3 surface expression, such that loss of responsiveness occurred with reduced CD3 cell-surface density. However, even thymocytes with very low CD3 expression were able to respond to triggering via CD3 under optimal conditions, indicating that the CD3 signal-transducing mechanism is functional on early thymic cells. Intracellular increases in Ca2+ concentrations induced via CD3, could effectively be inhibited by cross-linking of CD45 and CD3 on immature thymocytes. Although triggering via CD2 alone induced a strong Ca2+ flux, prolonged incubation with activating anti-CD2 antibodies made thymocytes refractory to subsequent triggering. Refractoriness was associated with partial loss of surface CD3 and CD3 zeta. Our results indicate that thymic surface Ag are differentially involved in the regulation of intracellular Ca2+ levels in immature as well as mature thymocytes.
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Mahmud, Dolores, and Damiano Rondelli. "Regulatory T Cells (Tregs) Can Be Isolated from G-CSF Mobilized PBSC after Monocyte Depletion and Inhibit Anti-Stem Cell T Cell Alloreactivity." Blood 112, no. 11 (November 16, 2008): 3477. http://dx.doi.org/10.1182/blood.v112.11.3477.3477.
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Abstract Small numbers of human CD4+CD25+FoxP3+ Tregs can be isolated from normal peripheral blood, thus their potential clinical application is limited. In this study we tested whether Granulocyte Colony-Stimulating Factor (G-CSF)-mobilized peripheral blood stem cells (PBSC) from healthy donors can represent a useful source of CD4+CD25+ cells with regulatory activity. We utilized antibodies conjugated to microbeads (Miltenyi Biotec Inc, Auburn, CA) to immunomagnetically separate the cells on a MidiMACS device (Miltenyi) and checked the purity after isolation by flow cytometry. Starting from on average 4.0±1.8 × 108 unseparated PBSC (n=3) we positively selected 2.4±0.8 × 106 CD34+ cells (>90% purity). We then utilized the CD34− cell fraction to isolate Tregs. Due to the large content of CD4dim monocytes in the initial cell product, we initially depleted the PBSC of CD14+ cells and then utilized a two-step process that includes a CD4+ cell negative selection (using a cocktail of biotin-conjugated antibodies against CD8, CD14, CD19, CD16, CD36, CD56, CD123, TCR g/δ and Glycophorin-A) followed by a positive selection of CD25+ cells (Treg isolation kit, Miltenyi). This process allowed us to obtain 1.2±1 × 106 CD4+CD25+ cells with a purity of >70%. Intracellular expression of FoxP3 was also detected in purified CD4+CD25+ cells by flow cytometry. Primary mixed leukocyte cultures (MLC) were performed with irradiated CD34+ cells isolated from PBSC and HLA mismatched blood CD3+ responders for 6 days and T cell response was measured by a 3H-thymidine uptake assay. Tregs isolated from PBSC were added to the MLC at 1:2 Treg:responder ratio to test their regulatory function. Control experiments were performed using CD4+CD25− cells. Addition of Tregs isolated from PBSC resulted in 76±17% inhibition of anti-CD34 T cell alloreactivity (cpm: 19000±530 vs 4590±1880) (n=3), while control CD4+CD25 neg cells did not show suppressive activity. These findings show that after isolation of CD34+ cells, adequate numbers of Tregs can be obtained from the CD34− cell fraction of PBSC by using a three-step process. In addition, since Tregs isolated from PBSC suppressed in-vitro T cell alloreactivity against CD34+ cells, these findings will prompt the design of pre-clinical studies to test the combination of PBSC-derived CD34+ cells and Tregs in HLA mismatched transplantation.
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Mittler, R. S., G. L. Schieven, P. M. Dubois, K. Klussman, M. P. O'Connell, P. A. Kiener, and V. Herndon. "CD45-mediated regulation of extracellular calcium influx in a CD4-transfected human T cell line." Journal of Immunology 153, no. 1 (July 1, 1994): 84–96. http://dx.doi.org/10.4049/jimmunol.153.1.84.
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Abstract Transfection of a CD4- Jurkat leukemic T cell line with the human wild-type CD4 gene resulted in the constitutive mobilization of calcium by these cells. The altered calcium phenotype was dependent on expression of a completely functional CD4 molecule on the cell surface. Transfectants receiving the vector alone or those in which a mutated CD4 gene lacking a functional Lck binding region failed to generate a constitutive calcium response. In addition, CD4 wild-type transfectants over time lost CD4 expression. These CD4- revertants failed to constitutively mobilize calcium. Treatment of CD4 wild-type transfected cells with either anti-CD45 mAb, EGTA, or PMA rapidly restored the cells to basal levels of intracellular calcium. Analysis of CD45 cross-linking on CD4+ and CD4- normal Jurkat lines demonstrated that CD4 expression was required for CD45-mediated inhibition of TCR induced calcium responses. CD45-mediated inhibition affected the duration of the response rather than its magnitude. These results, taken together with the observations obtained with CD4 transfected Jurkats suggested that influx of extracellular calcium was the predominant reason for the elevated levels of calcium within the cell. In support of this hypothesis, we could find no evidence of phosphorylated phospholipase C gamma 1 or constitutive inositol 1,4,5 trisphosphate generation in the CD4 wild-type transfected cells, yet both were detectable after anti-CD3 mAb-induced activation. Immunokinase assays of Lck and Fyn precipitated from untreated or anti-CD45-treated wild-type transfectants demonstrated that CD45 cross-linking dephosphorylated Lck and reduced its capacity to phosphorylate enolase. In contrast, neither Fyn autophosphorylation nor its activity was affected by CD45 cross-linking.
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Kraft, Daniel L., Agnieszka Czechowicz, and Irving L. Weissman. "Adult Human Hematopoietic Cells Differentiate into Mature T Cells Via a CD3-4+8- Intermediate within the Mouse Thymic Microenvironment; a New Model System for the Study of Human Thymocyte Development." Blood 106, no. 11 (November 16, 2005): 522. http://dx.doi.org/10.1182/blood.v106.11.522.522.
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Abstract Normal human T lymphocyte differentiation occurs within the thymus. We previously determined that human thymocyte precursors develop via a novel CD3-4+8- intermediate population, utilizing SCID-hu thymic grafts (Kraft, Weissman &Waller, JEM). We have recently worked to develop a more convenient and robust model of human hematopoiesis utilizing RAG2/Common gamma chain double knock out mice (RAG2 DKO) transplanted with hematopoietic stem and progenitor cells from adult human donors. Methods: Human peripheral blood derived CD34+ cells were obtained to >90% purity by magnetic bead positive selection following apheresis from healthy GCSF mobilized donors. 200,000-800,000 human CD34+ cells were injected intrahepatically into 0–2 day old RAG2/CG DKO pups following 2Gy x 2 of irradiation. At serial time points following intrahepatic transplantation, human CD45+ chimerism and donor derived phenotype was measured within the bone marrow, peripheral blood, spleen, liver, lymph node and thymus by flow cytometry. Thymuses were further analyzed utilizing antibodies to human CD3, CD4 and CD8. Results: Robust human thymopoeisis was observed in CD34+ transplanted mice. The degree of human engraftment increased with the number of CD34+ cells transplanted. Overall >70% of recipient mice successfully engrafted with >10% human CD45+ expression in analyzed tissues, with a mean of 63% of cells within the thymus being human derived. At earlier time points (4–6 weeks post transplant) the thymus of recipient mice were found to contain high fractions of immature CD45+ CD3-4-8- cells (making up 31–43% of human cells within the thymus) and the CD3-4+8- intermediate (22–30%) and CD4+8+ double positive (34–71%) populations with very rare mature CD3+4+8- or CD3+4-8+ T-cells (figure A). At later time points the fraction of immature CD3-4-8- and CD3-4+8- populations declined and increasing fractions of mature CD3+4+8- and CD3+4-8+ populations were identified (Figure B), in distributions similar to those found in normal human thymus. Conclusions: Human T-cells can differentiate within the mouse thymus derived from adult human CD34+ cells, and development appears to progress normally within a murine thymic microenvironment. The early developement of a CD3-4+8- intermediate suggests that T-cell development occurs via this population, unlike the CD3-4-8+ intermediate found in murine thymopoesis, suggesting that the pathway of human T-cell differentiation is intrinisic to the human thymocytes, and is independant of whether the thymic stroma is human (as found in SCID-hu) or murine (in Rag2 DKO). This robust model system enabling study of human thymopoesis utilizing hematopoietic stem cells from normal and diseased adults human donors may provide significant advantages for the study of human intrathymic T-cell differentiation and function in-vivo. Human thymocyte profile in the RAG2 DKO thymus A) 4 and B) 8 weeks following intrahepatic transplantation of human CD34+ cells. Right panels: CD3 profile of CD4+8− population reveals a CD3−4+8− population Human thymocyte profile in the RAG2 DKO thymus A) 4 and B) 8 weeks following intrahepatic transplantation of human CD34+ cells. Right panels: CD3 profile of CD4+8− population reveals a CD3−4+8− population
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Plum, J., M. De Smedt, MP Defresne, G. Leclercq, and B. Vandekerckhove. "Human CD34+ fetal liver stem cells differentiate to T cells in a mouse thymic microenvironment." Blood 84, no. 5 (September 1, 1994): 1587–93. http://dx.doi.org/10.1182/blood.v84.5.1587.bloodjournal8451587.
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Hematopoietic stem cells differentiate in the thymus to T cells along precisely defined intermediates. This process is thymic epithelium dependent and involves cytokines and cell-cell interactions between thymic stroma and T-cell precursors. Here we report that highly purified human CD34++ fetal liver stem cells differentiate to mature T cells, when seeded into isolated fetal thymic lobes of severe combined immunodeficient mice, and subsequently cultured in vitro. The human stem cells differentiate sequentially into CD4+CD8-CD3-, CD4+CD8+CD3-, CD4+CD8+CD3+, and finally, CD4+CD8-CD3+4 and CD4-CD8+CD3++ cells. Phenotypic analysis for additional maturation markers showed that these CD4 and CD8 single-positive thymocytes are fully maturate cells. By immunochemistry, human HLA-DR+ cells with a dendritic morphology could be detected. This novel chimeric human-mouse fetal thymus organ culture offers a tool to study human T-cell ontogeny in vitro and is a rapid and reliable test method for T-cell precursor activity of cultured or transfected human stem cells.
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Burchielli, Emanuela, Antonella Tosti, Loredana Ruggeri, Katia Perruccio, Claudia De Angelis, and Andrea Velardi. "Adoptive Therapy with T-Cell Precursors for Immune Reconstitution After Allogeneic Hematopoietic Stem Cell Transplantation." Blood 114, no. 22 (November 20, 2009): 3525. http://dx.doi.org/10.1182/blood.v114.22.3525.3525.
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Abstract Abstract 3525 Poster Board III-462 Recipients of allogeneic hematopoietic transplantation experience a slow reconstitution of donor-derived B and T cell number and function. This post-transplant period of immunodeficiency is associated with an increased risk of infection and malignant relapse. The developement of these complications notably correlates with the recovery of CD4+T cell subset. We proposed a strategy to enhances in vivo reconstitution by promoting donor-derived T cell development in the recipient's thymus. Recently Notch1-based ex-vivo system have been established to mature cord blood- or bone marrow-derived human HSCs into committed T-cell precursors. We used this system for the generation of T-cell precursors starting from G-CSF mobilized human HSCs. We cultured mobilized human CD34+ hematopoietic stem cells (HSCs) (2.5 × 105) in vitro on OP9 mouse stromal cells expressing the Notch 1 ligand Delta-like-1 (OP9-DL1) in the presence of rhFLT3-ligand (5ng/ml) and rhIL7 (5 ng/ml). After 6 weeks of co-culture we obtained a 3 log increase of human T-linage precursors of CD45RA+CD7high phenotype. Further co-colture (7-9 weeks) leed to the generation of CD4+ and CD8+ double-positive (DP) T cells and even mature CD4+ and CD8+ single positive (SP) ab-TCR lymphocytes. Experiments were designed in order to evaluate whether human CD45RA+CD7high T cell precursors could 1) engraft into NOD-SCID IL2 rg-/− mice 2) leed to in vivo expansion and maturation along T cell developmental pathway. Control mice were irradiated and transplanted with G-CSF-mobilized human CD34+ (dose 5×106 i.v.). 4 weeks after transplant more than 20% human CD45 positive cells engrafted in the bone marrow. Thymic engraftment occured at 8 weeks after transplant, with 80% human CD45 positive cells (thymic cellularity: 2.7×105 cells), mostly with T cell-immature phenotype of CD3-CD4-CD8 triple negative (95%) (TN) and CD4+CD8+double positive (5%) (DP). Co-transplant of CD45RA+CD7high T cell precursors (106 cells i.v.) along with CD34+HSC leed to an accelerated thymic engraftment (95% human CD45 positive cells; thymic cellularity 2.5 × 106 cells) already at 6 wks after transplant. Thymocytes were CD3-CD4-CD8 triple negative (51%) (TN) and CD4+CD8+double positive (DP) (42%) cells and at 8 weeks after transplant matured into CD3+CD4+ and CD3+CD8+ single positive (SP) T cells. Spectratyping analyses revealed a broad diversity of the T-cell receptor (TCR) repertoire. This occured in the complete absence of Graft versus Host Disease (GvHD) suggesting that adoptively transferred ex vivo-generated T-cell precursors developed into host-tolerant mature T cells. Ongonig experiment are needed to clarify the beneficial effect of adoptive immunotherapy with human T cell precursors on peripheral T cell reconstitution and control of infection in the humanized mouse system. We conclude that ex-vivo generation of human T-linage precursors is feasible from the G-CSF-mobilized HSCs and that can be succesfully tranfered in-vivo as a new strategie to enhance T-cell reconstitution after allogeneic HSCT with no risk of GvHD. Disclosures: No relevant conflicts of interest to declare.
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Зыблева, С. В. "Efficiency of the Use of Immunomodulatorsin Children with Recurrent Infections of the Organs of the Bronchopulmonary System." Рецепт, no. 5 (January 28, 2021): 762–74. http://dx.doi.org/10.34883/pi.2020.23.5.0012.
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Цель. Оценить показатели иммунной системы после применения иммуномодуляторов у детей с рецидивирующими инфекциями органов бронхолегочной системы.Материалы и методы. Обследовано 29 детей с частыми инфекциями верхних дыхательных путей, в течение года перенесших не менее 2 эпизодов болезни с поражением органов бронхолегочной системы: бронхит или пневмония. Изучали иммунофенотип лейкоцитов на основе моноклональных антител к СD3 (FITC), CD4 (FITC, PE), CD8 (PC-5, PE), CD56+16 (PE), CD11A (PE), CD14 (FITC), CD18 (FITC), CD19 (FITC), CD22 (FITC), CD25 (PC-5), CD28 (PC-5), CD40 (PE), CD45 (FITC,PC-5), CD71 (FITC), CD95 (PE), CD154 (PE), HLA-DR (PC-5). Показатели оценивали перед, через 10 дней и через 2 месяца после курса иммунореабилитации.Результаты и обсуждение. Выявлено отсутствие значимых отличий показателей CD3+CD4+CD95+ Т-хелперов от уровня контрольной группы у детей с рецидивирующими заболеваниями органов бронхолегочной системы. Уровень нейтрофилов, экспрессирующих рецепторы адгезии CD11a+ и CD18+, достиг в результате применения иммуномодуляторов уровня показателей контрольной группы, что указывает на положительный эффект проводимой терапии. Выявлено повышение липополисахаридсвязывающей способности Т- и В-лимфоцитов и отсутствие статистически значимых отличий от контроля субпопуляций лимфоцитов, экспрессирующих рецепторы к липополисахариду CD19+LPS+ и CD3+LPS+, у детей с рецидивирующими заболеваниями органов бронхолегочной системы через 10 дней и 2 месяца после курса иммунореабилитации. Обнаружен рост силы корреляционной связи LPS+CD19+ с LPS+CD3+ через10 дней с rs=0,47 до rs=0,7, с некоторым снижением к 2 месяцам от начала иммунореабилитации до rs=0,57.Выводы. Курс иммунореабилитации способствует адаптации активационных процес-сов системы иммунитета. Динамическое определение экспрессии рецепторов адгезии CD11a+ и CD18+ нейтрофилов, липополисахаридсвязывающей способности лимфоцитов, CD3+CD8+CD28+ активированных Т-лимфоцитов, CD3+CD4+CD95+ Т-хелперов, CD154+лимфоцитов при иммунореабилитации в период ремиссии может служить лабораторным критерием эффективности иммунореабилитации и использоваться для формирования группы риска по рецидивирующей инфекции у детей данного профиля. Purpose. To assess the parameters of the immune system after the use of immunomodulators in children with recurrent infections of the bronchopulmonary system.Materials and methods. We examined 29 children with frequent infections of the upper respiratory tract, who had suffered at least 2 episodes of the disease with the damage to the organs of the bronchopulmonary system during the year: bronchitis or pneumonia. We have studied the leukocyte immune phenotype based on monoclonal antibodies to СD3 (FITC), CD4 (FITC, PE), CD8 (PC-5, PE), CD56+16 (PE), CD11A (PE), CD14 (FITC), CD18 (FITC), CD19 (FITC), CD22 (FITC), CD25 (PC- 5), CD28 (PC-5), CD40 (PE), CD45 (FITC, PC-5), CD71 (FITC), CD95 (PE), CD154 (PE), HLA-DR (PC-5).The indicators were assessed before the course of immune rehabilitation, in 10 days and 2 months after it.Results and discussion. The absence of significant differences in the indicators of CD3+CD4+CD95+ T-helpers from the level in the control group of children with recurrent diseases of the bronchopulmonarysystemwasrevealed.The levelofneutrophilsthatexpresstheadhesionreceptors CD11a+ and CD18+ reached the level of the control group, as a result of the use of immunomodulators, which indicates a positive effect of the therapy. The increase of the LPS-binding capacity of T- and B-lymphocytes and the absence of statistically significant differences from the control of subpopulations of lymphocytes that express the receptors for lipopolysaccharide CD19+LPS+ and CD3+LPS+ were revealed in children with recurrent diseases of the bronchopulmonary system in 10 days and 2 months after the course of immunorehabilitation. The increase of the strength of thecorrelation relationship LPS+CD19+ with LPS+CD3+ was determined in 10 days from r =0,47 to r =0,7,s swith some decrease by 2 months from the beginning of immunorehabilitation to rs=0,57.Conclusion. The course of immunorehabilitation contributes to the adaptation of the activationprocesses of the immune system. Dynamic determination of the expression of adhesion receptors CD11a+ and CD18+ neutrophils, LPS-binding capacity lymphocytes, CD3+CD8+CD28+ activated T-lymphocytes, CD3+CD4+CD95+ T-helper cells, CD154+ lymphocytes in immunorehabilitation during the period of rehabilitation can serve as a laboratory criterion of immunorehabilitation efficiency, and it can be used to form the risk group of children with recurrent infection.
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Tamiolakis, Demetrio, Ioannis Venizelos, Athanasia Kotini, Sylva Nikolaidou, and Nikolaos Papadopoulos. "Prevalence of CD8/CD4 Ratio in the Fetal Thymic Parenchyme in Down’s Syndrome." Acta Medica (Hradec Kralove, Czech Republic) 46, no. 4 (2003): 179–82. http://dx.doi.org/10.14712/18059694.2019.30.
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Aim: The maturation of most T- lymphocyte precursors takes place within the meshwork of thymic epithelial cells. Different steps of this process can be defined by immunologic phenotyping. The prothymocytes are positive for the terminal deoxynucleotidyl transferase (TdT) and give rise to cortical thymocytes, which express CD1, CD2, CD3, CD5, and both CD4 and CD8. These CD4 and CD8 double-positive cortical thymocytes differentiate into two lineages: CD4+ or CD8+ lymphocytes of the thymic medulla, by the tenth week of gestation. Our study points towards the determination of the CD8 cytotoxic/suppressor capacity of the fetal thymus in Down’s syndrome. Experimental design: A quantitative comparison of T-lymphocytes (CD3, CD4, and CD8) in the thymic parenchyme in embryos after voluntary abortion during 2nd trimester of gestation and embryos with Down’s syndrome, respectively, was performed. Results: Our results showed: 1) A statistically significant depletion in the total number of T-cells (CD3 positive) in the cases of embryos with Down’s syndrome over those after voluntary abortion, during the second trimester of gestation (p<0.0001, t-test). 2) A significant difference in the CD8/CD4 ratio in the cases of embryos with Down’s syndrome, during the second trimester of gestation which was numerically stronger with the progress of fetal development (20th week: p<0.025; 24th week: p<0.01, chi-square). Conclusions: The occurrence of increased CD8/CD4 ratio in the cases with Down’s syndrome, in the second trimester of gestation, underlines the cytotoxic / suppressor property of the thymus in the affected fetuses.
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Blom, Bianca, Martie C. M. Verschuren, Mirjam H. M. Heemskerk, Arjen Q. Bakker, Ellen J. van Gastel-Mol, Ingrid L. M. Wolvers-Tettero, Jacques J. M. van Dongen, and Hergen Spits. "TCR Gene Rearrangements and Expression of the Pre-T Cell Receptor Complex During Human T-Cell Differentiation." Blood 93, no. 9 (May 1, 1999): 3033–43. http://dx.doi.org/10.1182/blood.v93.9.3033.
Full textAbstract:
Abstract Recent studies have identified several populations of progenitor cells in the human thymus. The hematopoietic precursor activity of these populations has been determined. The most primitive human thymocytes express high levels of CD34 and lack CD1a. These cells acquire CD1a and differentiate into CD4+CD8+ through CD3−CD4+CD8− and CD3−CD4+CD8+β− intermediate populations. The status of gene rearrangements in the various TCR loci, in particular of TCRδ and TCRγ, has not been analyzed in detail. In the present study we have determined the status of TCR gene rearrangements of early human postnatal thymocyte subpopulations by Southern blot analysis. Our results indicate that TCRδ rearrangements initiate in CD34+CD1a− cells preceding those in the TCRγ and TCRβ loci that commence in CD34+CD1a+ cells. Furthermore, we have examined at which cellular stage TCRβ selection occurs in humans. We analyzed expression of cytoplasmic TCRβ and cell-surface CD3 on thymocytes that lack a mature TCRβ. In addition, we overexpressed a constitutive-active mutant of p56lckF505 by retrovirus-mediated gene transfer in sequential stages of T-cell development and analyzed the effect in a fetal thymic organ culture system. Evidence is presented that TCRβ selection in humans is initiated at the transition of the CD3−CD4+CD8− into the CD4+CD8+β− stage.
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Blom, Bianca, Martie C. M. Verschuren, Mirjam H. M. Heemskerk, Arjen Q. Bakker, Ellen J. van Gastel-Mol, Ingrid L. M. Wolvers-Tettero, Jacques J. M. van Dongen, and Hergen Spits. "TCR Gene Rearrangements and Expression of the Pre-T Cell Receptor Complex During Human T-Cell Differentiation." Blood 93, no. 9 (May 1, 1999): 3033–43. http://dx.doi.org/10.1182/blood.v93.9.3033.409k39_3033_3043.
Full textAbstract:
Recent studies have identified several populations of progenitor cells in the human thymus. The hematopoietic precursor activity of these populations has been determined. The most primitive human thymocytes express high levels of CD34 and lack CD1a. These cells acquire CD1a and differentiate into CD4+CD8+ through CD3−CD4+CD8− and CD3−CD4+CD8+β− intermediate populations. The status of gene rearrangements in the various TCR loci, in particular of TCRδ and TCRγ, has not been analyzed in detail. In the present study we have determined the status of TCR gene rearrangements of early human postnatal thymocyte subpopulations by Southern blot analysis. Our results indicate that TCRδ rearrangements initiate in CD34+CD1a− cells preceding those in the TCRγ and TCRβ loci that commence in CD34+CD1a+ cells. Furthermore, we have examined at which cellular stage TCRβ selection occurs in humans. We analyzed expression of cytoplasmic TCRβ and cell-surface CD3 on thymocytes that lack a mature TCRβ. In addition, we overexpressed a constitutive-active mutant of p56lckF505 by retrovirus-mediated gene transfer in sequential stages of T-cell development and analyzed the effect in a fetal thymic organ culture system. Evidence is presented that TCRβ selection in humans is initiated at the transition of the CD3−CD4+CD8− into the CD4+CD8+β− stage.