Infiltration of Canonical Vγ4/Vδ1 γδ T Cells in an Adriamycin-Induced Progressive Renal Failure Model

Abstract

We have previously reported an infiltration of renal interstitial γδ T cells in Adriamycin-induced progressive glomerulosclerosis in the rat kidney. The TCR repertoire and sequences used by these γδ T cells have now been studied. Two injections of Adriamycin 14 days apart caused segmental glomerulosclerosis, massive interstitial infiltration of mononuclear cells, and end-stage renal failure. Flow cytometry of lymphocyte subpopulations with Abs to CD3, the γδ TCR, and the αβ TCR showed that γδ 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) and creatinine (r = 0.62; p < 0.01) correlated significantly with the presence of γδ T cells. TCR Vγ repertoire analysis by RT-PCR and Southern blotting showed that Vγ2 was the dominant subfamily in lymph nodes, whereas Vγ4 became the predominant subfamily in advanced stages of the rat Adriamycin-treated kidney. Sequencing of the Vγ4-Jγ junctional region showed an invariant sequence. The amino acid sequence of the junctional region of the Vγ4 TCR was the same as the reported mouse canonical Vγ4 TCR sequence. Analysis of the kidney Vδ repertoire showed dominant expression of Vδ1, and sequencing again revealed the selective expression of a canonical Vδ1 gene. Semiquantitative RT-PCR for cytokine gene expression showed that γδ T cells from the kidneys expressed TGF-β, but not IL-4, IL-10, or IFN-γ. These results suggest that the predominant γδ T cells in the Adriamycin kidney use an invariant Vγ4/Vδ1 receptor.

Depletion of γδ T Cells Exacerbates Murine Adriamycin Nephropathy

“…All in-frame V␦1 sequences from the AN kidney used a canonical V␦1J␦2C␦ junctional sequence (Table 3). This canonical V␥6/V␦1 junctional sequence is identical to those expressed on fetal thymocytes and the same as we found in rat AN kidney (12). These results are matched with the sequence reported in a previously published orchitis model (11).…”

“…Our initial observation in human IgA nephritis that ␥␦ T cells are associated with worse disease is now revised by our findings of a regulatory phenotype of ␥␦ T cells in rat AN and HN and now in murine AN, where depletion is associated with worsening of disease (12,13). This difference may in part be explained by the expansion of a subset of ␥␦ T cells in response to local inflammation then providing a protective or regulatory role.…”

“…We previously showed this in rat Adriamycin nephropathy (AN), where there is expression of a highly restricted canonical V␥6/V␦1 TCR (12), and found a similar TCR restriction in Heymann nephritis (HN) (13). In both animal models and human glomerulonephritis, the expression of ␥␦ T cells is associated with disease severity (12)(13)(14). The regulatory role of ␥␦ T cells was examined by Rosenkranz et al (15) using knockout mice.…”

Detection of Cell Surface Ligands for the γδ TCR Using Soluble TCRs

“…Moreover, previous experiments have already implied that the unidentified ligands of these three ␥␦ TCRs are inducible host molecules: for the V␥6/ V␦1 TCR, a molecule induced in several different tissues during inflammation evoked by bacterial infection (38,42), by autoimmune attack (27), and by toxic drugs (20); for the V␥5/V␦1 TCR, a molecule inducible on stressed keratinocytes (4); and for the V␥1/V␦6.3 TCR, an autologous molecule most likely expressed by certain tumor cells (2,55). Our finding that ␥␦ TCRs can be used to detect their own ligands will likely prove useful in the full characterization of these ligands, and may as well identify the cells with which ␥␦ T cells and their subsets interact.…”

Interleukin-18 binding protein therapy is protective in adriamycin nephropathy

“…Macrophages are considered possible effectors of disease in AN, and a reduction in kidney injury has been demonstrated after partial macrophage depletion (46) or neutralization of macrophage effector pathways (3,50).…”