Publikation: Reduced CTLA-4 protein and messenger RNA expression in umbilical cord blood T lymphocytes
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Objective: A favorable incidence and severity of graft-vs-host disease is observed in patients transplanted with banked, unrelated, HLA-mismatched umbilical cord blood (UCB) grafts, while the incidence of malignant relapse remains low. CTLA-4 mediates negative T-cell signaling and may contribute to the development of allogeneic tolerance. In this study, we compared protein and mRNA expression of CTLA-4 in stimulated UCB and adult peripheral blood T cells.
Materials and Methods: T cells were isolated from UCB and adult peripheral blood and stimulated with anti-CD3 and anti-CD28 monoclonal antibodies. Cells were immunostained and analyzed by flow cytometry for both surface and intracellular expression of CTLA-4 in the presence and absence of cyclosporin A, and kinetics of CTLA-4 expression compared. CTLA-4 mRNA expression was measured using quantitative real-time polymerase chain reaction. NFAT1 protein levels were measured by Western blot analysis.
Results: These studies demonstrate reduced surface and intracellular expression of CTLA-4 in stimulated UCB T cells compared to adult controls. Furthermore, reduced CTLA-4 protein expression in UCB T cells was noted to be in part transcriptionally regulated, as CTLA-4 mRNA levels also were significantly lower. Reduced CLTA-4 expression by UCB T cells followed the kinetics of delayed and reduced expression of the transcription factor NFAT1 by UCB T lymphocytes during primary stimulation. Moreover, cyclosporin A, which is known to modulate NFAT activation, reduced CTLA-4 protein expression in adult and UCB T cells.
Conclusion: Reduced expression of the key regulatory proteins CTLA-4 and NFAT-1 may contribute to favorable UCB T lymphocyte allogeneic responses.
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MILLER, Robin E., John D. FAYEN, Shaden F. MOHAMMAD, Kevin R. STEIN, Suzanne KADEREIT, Kathleen DAUM-WOODS, R. Michael SRAMKOSKI, James W. JACOBBERGER, Dennis TEMPLETON, Susan B. SHURIN, Mary J. LAUGHLIN, 2002. Reduced CTLA-4 protein and messenger RNA expression in umbilical cord blood T lymphocytes. In: Experimental Hematology. 2002, 30(7), pp. 738-744. ISSN 0301-472X. Available under: doi: 10.1016/S0301-472X(02)00831-7BibTex
@article{Miller2002Reduc-7303, year={2002}, doi={10.1016/S0301-472X(02)00831-7}, title={Reduced CTLA-4 protein and messenger RNA expression in umbilical cord blood T lymphocytes}, number={7}, volume={30}, issn={0301-472X}, journal={Experimental Hematology}, pages={738--744}, author={Miller, Robin E. and Fayen, John D. and Mohammad, Shaden F. and Stein, Kevin R. and Kadereit, Suzanne and Daum-Woods, Kathleen and Sramkoski, R. Michael and Jacobberger, James W. and Templeton, Dennis and Shurin, Susan B. and Laughlin, Mary J.} }
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