The aminonucleoside of puromycin (PAN) induces nephrotic syndrome in rats. We studied the tubulointerstitial cellular (TIC) infiltrate previously unrecognized in this model. Rats received one i.p. injection of PAN (15 mg/100 g) and were sacrificed at 1, 3, 4, 5, 7, 14, 20 and 28 days. Frozen kidney sections and peripheral blood cells were stained with a panel of anti-rat monoclonal antibodies and quantitated by epifluorescence microscopy. An increase in Ia+ cells (60/1000 TIC) (P less than 0.001) and OX42+ macrophages (MO) (18/1000 TIC) (P less than 0.05) were observed on day 5. On day 7 the infiltrate consisted of OX19+ T-lymphocytes (29/1000 TIC) (P less than 0.001) and OX42+ MO (68/1000 TIC) (P less than 0.001). The majority of the lymphocytes expressed the OX8 cytotoxic T cell marker (23/1000 TIC) (P less than 0.001). The severe mixed cellular lesion present on day 14 was dominated by OX42+ MO (113/1000 TIC) (P less than 0.001). With resolution of proteinuria on days 20 and 28, the infiltrate decreased, although OX42+ MO persisted on day 28 (46/1000 TIC) (P less than 0.001). The severity of the cellular lesion correlated with the degree of albuminuria (r = 0.57 to 0.81 for the antibody panel). Expression of Ia antigens by proximal tubular epithelial cells markedly decreased during peak proteinuria but normalized by day 28. Increased deposition of C3 and IgG was not detected. Reversible tubulointerstitial nephritis develops in PAN-treated rats and may be a consequence of severe proteinuria.