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 Blood, 1 April 2008, Vol. 111, No. 7, pp. 3675-3683.
Prepublished online as a Blood First Edition Paper on February 4, 2008; DOI 10.1182/blood-2008-01-130146.

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IMMUNOBIOLOGY

Rabbit ATG but not horse ATG promotes expansion of functional CD4+CD25highFOXP3+ regulatory T cells in vitro

Xingmin Feng1, Sachiko Kajigaya1, Elena E. Solomou1, Keyvan Keyvanfar1, Xiuli Xu2, Nalini Raghavachari2, Peter J. Munson3, Thomas M. Herndon1, Jichun Chen1, and Neal S. Young1

1 Hematology Branch, National Heart, Lung, and Blood Institute (NHLBI), National Institutes of Health (NIH), Bethesda, MD; 2 Genomics Core Facility, Pulmonary and Vascular Medicine Branch, NHLBI, NIH, Bethesda, MD; and 3 Mathematical and Statistical Computing Laboratory, Division of Computational Bioscience, Center for Information Technology, NIH, Bethesda, MD

Regulatory T cells (Treg) play important roles in suppressing immune responses and maintaining tolerance. Rabbit antithymocyte globulin (rATG) and horse ATG (hATG) are widely used in the treatment of immune-mediated syndromes, but their effects on Treg are unknown. We show here that in vitro culture of normal human peripheral blood mononuclear cells (PBMCs) with a low-dose rATG resulted in marked expansion of functional Treg by converting CD4+CD25– T cells to CD4+CD25+ T cells. hATG did not expand but rather decreased Treg. Immuno-blot showed increased expression of FOXP3 and NFAT1 in CD4+CD25– and CD4+CD25+ T cells exposed to rATG. PBMCs treated with rATG displayed increased interleukin-10 in culture supernatants than those treated with hATG. Furthermore, rATG and hATG showed differences in their potential to stimulate CD4+ T cells as examined using different activation markers. Microarray revealed that rATG induced markedly different gene-expression patterns in PBMCs, compared with hATG-treated or untreated PBMCs. Our findings indicate that rATG expanded Treg, probably through transcriptional regulation by enhanced NFAT1 expression, in turn conferring CD4+CD25– T cell FOXP3 expression and regulatory activity. The therapeutic effects of rATG may occur not only because of lymphocyte depletion but also enhanced Treg cell number and function.


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