Abstract:
BACKGROUND:Alloreactive immune responses may engage both direct and indirect antigen allorecognition. This study focuses on T-cell receptor (TCR) beta-chain usage by in vitro generated alloreactive CD4+ T cells involved in direct allorecognition pathway. METHODS AND RESULTS:We have established Lewis anti-Brown Norway rat CD4+ T-cell lines and confirmed their reactivities against cell-surface, but not soluble, alloantigens. TCR Vbeta-specific reverse-transcriptase polymerase chain reaction detected all 22 Vbeta genes in these cell lines at all stages, regardless of the lengths of in vitro stimulation. By using spectrotyping, we found that Vbeta complementarity determining region (CDR)3 length distribution pattern altered dramatically after repeated allostimulation. Such a skewed CDR3 distribution occurred in most Vbeta genes without any obvious preference, indicating that expansion occurred in all Vbeta expressing cells by allostimulation. Fluorescence-activated cell sorter analysis of Vbeta expression in alloreactive CD4+ T-cell lines with anti-Vbeta-specific monoclonal antibodies showed, quantitatively, similar percentages of individual Vbeta-expressing cells in the alloreactive pool after repeated allostimulation. To test whether preferential TCR Vbeta gene usage occurred in "high responder" cells, we sorted CD4+ T cells that underwent three or more divisions from primary mixed leukocyte reactions. Unlimited Vbeta usage with CDR3 alterations was observed, as in unsorted alloreactive T cells. CONCLUSION:TCR Vbeta gene usage in directly alloreactive CD4+ T-cell population is unrestricted. Clonal expansion occurs in all Vbeta expressing T cells by allostimulation.
journal_name
Transplantationjournal_title
Transplantationauthors
Zhai Y,Kupiec-Weglinski JWdoi
10.1097/01.TP.0000046939.45400.43subject
Has Abstractpub_date
2003-02-27 00:00:00pages
514-21issue
4eissn
0041-1337issn
1534-6080journal_volume
75pub_type
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