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Reduced CTLA-4 protein and messenger RNA expression in umbilical cord blood T lymphocytes

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2002

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Miller, Robin E.
Fayen, John D.
Mohammad, Shaden F.
Stein, Kevin R.
Daum-Woods, Kathleen
Sramkoski, R. Michael
Jacobberger, James W.
Templeton, Dennis
Shurin, Susan B.

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Experimental Hematology. 2002, 30(7), pp. 738-744. ISSN 0301-472X. Available under: doi: 10.1016/S0301-472X(02)00831-7

Zusammenfassung

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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570 Biowissenschaften, Biologie

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ISO 690MILLER, 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)00831-7
BibTex
@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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