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 Blood, 29 October 2009, Vol. 114, No. 18, pp. 3947-3955.
Prepublished online as a Blood First Edition Paper on August 27, 2009; DOI 10.1182/blood-2009-03-211896.

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TRANSPLANTATION

The human alloreactive CD4+ T-cell repertoire is biased to a Th17 response and the frequency is inversely related to the number of HLA class II mismatches

Nicolle H. R. Litjens1, Jacqueline van de Wetering1, Nicole M. van Besouw2, and Michiel G. H. Betjes1

1 Department of Internal Medicine, Division of Nephrology, and 2 Laboratory of Transplantation, Division of Renal Transplantation, Erasmus Medical Center, Rotterdam, The Netherlands

Estimates of precursor frequency and assessment of functional characteristics of alloreactive CD4+ T cells are all biased by the need for long-term culture. In this study, direct visualization of human alloreactive CD4+ T cells on the single-cell level was achieved using cell surface expression of CD154 as a tool for identification. The average frequency of alloreactive CD154+CD4+ T cells among peripheral blood CD4+ T cells was 0.1%, with half of the cells displaying a naive phenotype. The proliferation capacity and expression of cytokines after allogeneic stimulation resided in these CD154+CD4+ T cells. The repertoire of alloreactive CD4+ T cells was biased to a Th17 response, and on average 24% of alloreactive CD154+CD4+ memory T cells produced interleukin-17 (IL-17) after polyclonal stimulation. Unexpectedly, mixed cell cultures from human leukocyte antigen (HLA)–identical donors also generated alloreactive CD154+CD4+ T cells and yielded the highest frequency compared with HLA-nonidentical combinations. Therefore, reactivity to minor histocompatibility antigens between HLA-identical subjects appears to be relatively common. Alloreactive HLA-identical T cells did not proliferate or express cytokines, but were driven to proliferation in the presence of exogenous IL-2.


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