Umbilical cord blood (UCB) stem cells have emerged as novel treatment for patients requiring allogeneic grafting. The incidence and severity of graft-versus-host disease (GVHD) after UCB transplantation in these early clinical trials compares favorably to that observed in recipients of matched unrelated donors (MUD) or partially mismatched family member allogeneic grafts. It has been shown that stimulated lymphocytes from UCB have reduced production of cytokines including IFN-gamma and TNF-alpha, known to play an important role in GVHD pathophysiology. This laboratory has investigated the molecular mechanisms underlying this reduced cytokine production by analyzing expression of nuclear factor of activated T cells-1 (NFAT1) in UCB T cells. The applicants detected no constitutive expression of NFAT1 protein in non-stimulated UCB T cells, and although NFAT1 expression in UCB T cells was upregulated after prolonged (40 hr) T cell stimulation, it was only partially upregulated when compared to adult controls. These observations of minimal NFAT1 expression after stimulation correlated with reduced cytoplasmic IFN-gamma and TNF-alpha production in UCB T cells studied simultaneously. The first hypothesis underlying the work outlined in this application is that gene regulation of NFAT1, a central transcription factor involved in cytokine gene expression, differs between neonatal and adult T cells. Secondly, they hypothesize that that these fundamental differences in NFAT1 regulation between neonatal and adult T cells will elucidate underlying regulatory pathways mediating hyporesponsiveness of UCB T cells. To test these hypotheses, they will [Aim 1] compare in detail expression kinetics and gene regulation of NFAT1 in UCB and adult T cells. Then [Aim 2] in vitro studies will be performed to compare NFAT1 protein levels with surface expression of immunomodulatory molecules including CD40L, CTLA-4, and FasL on UCB T cells, and cytoplasmic expression of IFN-gamma and TNF-alpha, in the presence or absence of cyclosporine or FK506 (tacrolimus). Finally, [Aim 3] NFAT1 protein levels and surface expression T cells of immunomodulatory molecules will be compared in donor UCB and HLA-matched sibling T cells emerging during immune recovery after transplant. These studies may provide further insight into the regulation and expression of above described genes during neonatal and adult life. Moreover, since NFAT1 regulates the gene expression of cytokines and immunomodulatory molecules known to mediate GVHD, their studies may reveal new therapeutic approaches to the treatment and/or prophylaxis of GVHD.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI047289-04
Application #
6632266
Study Section
Special Emphasis Panel (ZRG1-ET-1 (02))
Program Officer
Kehn, Patricia J
Project Start
2000-04-01
Project End
2005-03-31
Budget Start
2003-04-01
Budget End
2004-03-31
Support Year
4
Fiscal Year
2003
Total Cost
$264,250
Indirect Cost
Name
Case Western Reserve University
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
077758407
City
Cleveland
State
OH
Country
United States
Zip Code
44106
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Lesniewski, M L; Haviernik, P; Weitzel, R P et al. (2008) Regulation of IL-2 expression by transcription factor BACH2 in umbilical cord blood CD4+ T cells. Leukemia 22:2201-7
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Kaminski, Beth A; Kadereit, Suzanne; Miller, Robin E et al. (2003) Reduced expression of NFAT-associated genes in UCB versus adult CD4+ T lymphocytes during primary stimulation. Blood 102:4608-17
Kadereit, Suzanne; Junge, Gwendolyn R; Kleen, Thomas et al. (2003) Deficient IFN-gamma expression in umbilical cord blood (UCB) T cells can be rescued by IFN-gamma-mediated increase in NFATc2 expression. J Clin Immunol 23:485-97
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