INFILTRATION OF CANONICAL V(4/V*1 (*T CELLS IN ADRIAMYCIN-INDUCED PROGRESSIVE GLOMERULOSCLEROSIS

Ando T, ^1,2 Watson D, ¹ Wu H, ¹ Hirano T, ² Hirakata H, ² Fujishima M, ² Knight JF¹

^¹Centre for Kidney Research, New Children's Hospital, Westmead. ^² Second Dept of Internal Medicine, Kyushu University, Fukuoka, Japan

We have previously reported an infiltration of renal interstitial (*T cells in adriamycin-induced progressive glomerulosclerosis (ADR) in the rat kidney. The T-cell receptor (TCR) repertoire and sequences of these (*T cells have now been studied. Two injections of adriamycin (2.5 mg/kg BW) 14 days apart given to male Sprague-Dawley rats caused segmental glomerulosclerosis, massive interstitial infiltration of mononuclear cells and end stage renal failure. The rats were sacrificed at 18 to 22 weeks after injection. Kidneys and lymph nodes were minced and mononuclear cells isolated on a density gradient. Flow cytometry of lymphocyte subpopulations with antibodies to CD3, (*TCR and "$TCR. showed that (*T cells as a proportion of CD3+ cells were increased in ADR kidneys (8.5 2.8%) but not in lymph nodes (1.4 0.5%). A semiquantitative score of glomerular damage(r=0.65, p ‹0.01) and creatinine (r=0.62, p‹0.01) correlated significantly with the presence of (*T cells. TCR V( repertoire analysis by RT-PCR and Southern blotting showed that V(2 was the dominant subfamily in lymph nodes whereas V(4 became the predominant subfamily in advanced stage rat ADR kidney. Sequencing of the V(4-J( junctional region showed an invariant sequence. The amino acid sequence of the junctional region of the V(4 TCR was exactly the same as the reported mouse canonical V(4 TCR sequence, although there are some nucleotide substitutions between rat and mouse. Analysis of kidney V* repertoire showed dominant expression of V*1, and sequencing again showed a sole canonical gene. These results suggest that the predominant (*T cells in the ADR kidney use an invariant V(4/V*1 receptor. This is the first report showing a significant infiltration of canonical (*T cells in renal disease. We speculate that this V(4/V*1 (*T cell subpopulation selectively recognises an unknown common ligand related to chronic inflammatory interstitial tissue damage.

Presented at the Annual Scientific Meeting of the Australian and New Zealand Society of Nephrology, Melbourne, March 2000