Infiltration of canonical V gamma 4/V delta 1 gamma delta T cells in an adriamycin-induced progressive renal failure model

We have previously reported an infiltration of renal interstitial gamma del ta T cells in Adriamycin-induced progressive glomerulosclerosis in the rat kidney. The TCR repertoire and sequences used by these gamma delta T cells have now been studied. Two injections of Adriamycin 14 days apart caused se gmental glomerulosclerosis, massive interstitial infiltration of mononuclea r cells, and end-stage renal failure. Flow cytometry of lymphocyte subpopul ations with Abs to CD3, the gamma delta TCR, and the alpha beta TCR showed that gamma delta T cells as a proportion of CD3(+) cells were increased in Adriamycin-treated kidneys (8.5 +/- 5.4%), but not in lymph nodes (1.3 +/- 0.4%). A semiquantitative score of glomerular damage (r = 0.65; p < 0.01) a nd creatinine (r = 0.62; p < 0.01) correlated significantly with the presen ce of gamma delta T cells. TCR V gamma repertoire analysis by RT-PCR and So uthern blotting showed that V gamma2 was the dominant subfamily in lymph no des, whereas V gamma4 became the predominant subfamily in advanced stages o f the rat Adriamycin-treated kidney. Sequencing of the V gamma4-J gamma jun ctional region showed an invariant sequence. The amino acid sequence of the junctional region of the V gamma4 TCR was the same as the reported mouse c anonical V gamma4 TCR sequence. Analysis of the kidney V delta repertoire s howed dominant expression of V delta1, and sequencing again revealed the se lective expression of a canonical V delta1 gene. Semiquantitative RT-PCR fo r cytokine gene expression showed that gamma delta T cells from the kidneys expressed TGF-beta, but not IL-4, IL-10, or IFN-gamma. These results sugge st that the predominant gamma delta T cells in the Adriamycin kidney use an invariant V gamma4/V delta1 receptor.