The Journal of Experimental Medicine
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Published 18 April 2005. doi:10.1084/jem.20042398
Rockefeller University Press, 0022-1007 $8.00
JEM, Volume 201, Number 8, 1333-1346
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ARTICLE

 Single cell analysis shows decreasing FoxP3 and TGFß1 coexpressing CD4+CD25+ regulatory T cells during autoimmune diabetes

Shannon M. Pop1, Carmen P. Wong2, Donna A. Culton2, Stephen H. Clarke2, and Roland Tisch1,2

1 Curriculum in Oral Biology, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599
2 Department of Microbiology and Immunology, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599

CORRESPONDENCE Roland Tisch: rmtisch{at}med.unc.edu

Natural CD4+CD25+ regulatory T (CD4+CD25+ T reg) cells play a key role in the immunoregulation of autoimmunity. However, little is known about the interactions between CD4+CD25+ T reg cells and autoreactive T cells. This is due, in part, to the difficulty of using cell surface markers to identify CD4+CD25+ T reg cells accurately. Using a novel real-time PCR assay, mRNA copy number of FoxP3, TGFß1, and interleukin (IL)-10 was measured in single cells to characterize and quantify CD4+CD25+ T reg cells in the nonobese diabetic (NOD) mouse, a murine model for type 1 diabetes (T1D). The suppressor function of CD4+CD25+CD62Lhi T cells, mediated by TGFß, declined in an age-dependent manner. This loss of function coincided with a temporal decrease in the percentage of FoxP3 and TGFß1 coexpressing T cells within pancreatic lymph node and islet infiltrating CD4+CD25+CD62Lhi T cells, and was detected in female NOD mice but not in NOD male mice, or NOR or C57BL/6 female mice. These results demonstrate that the majority of FoxP3-positive CD4+CD25+ T reg cells in NOD mice express TGFß1 but not IL-10, and that a defect in the maintenance and/or expansion of this pool of immunoregulatory effectors is associated with the progression of T1D.

Abbreviations used: Ab, antibody; CD62Lhi T cells, CD4+CD25+CD62Lhi T cells; CD62Llo T cells, CD4+CD25+CD62Llo T cells; CTLA-4, cytotoxic T lymphocyte–associated antigen 4; GITR, glucocorticoid-induced TNF receptor gene; NOD, nonobese diabetic; PLN, pancreatic LN; rh, recombinant human; T reg, regulatory T; T1D, type 1 diabetes.


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