Quantitating the severity of proximal tubular brush border injury by a simple direct binding radioimmunoassay.

The purpose of this investigation was: (1) to establish a simplified radioimmunoassay (RIA) for quantitating renal tubular epithelial antigens (RTE) in urine; (2) to ascertain whether urine RTE concentrations as measured by this technique correlate with the severity of acute nephrotoxic and ischemic injury; and (3) to ascertain whether increased urinary RTE is a specific marker of renal tubular injury. A direct binding RTE RIA was established using 125I labelled anti-RTE antibody and 10% polyethylene glycol to separate bound from unbound anti-RTE 125I. This RIA is simpler than previously described RTE assay methods since: (1) double antibody separation techniques are eliminated; (2) RTE antigen purification from crude proximal tubular fragments is no longer necessary; and (3) immunoglobulin G(IgG) rather than more radiosensitive RTE is used as the radioligand. To test the utility of this assay as a marker of acute tubular injury anesthetized rats were subjected to graded nephrotoxic (HgCl2: 0--20 mg/kg) or bilateral renal ischemic (0--32 min) insults. Glomerular filtration rates (GFR) (clearance iothalamate 125I) and RTE concentrations were measured sequentially. Post-renal injury, RTE concentrations rose above control values and the degree of elevation strongly correlated with the severity of GFR depression (r = 0.72--0.81; p less than 0.02--0.05). The source of this increased urinary RTE was the proximal tubule since brush border loss was demonstrated histologically and because no RTE could be detected in serum. Rats whose GFRs were acutely depressed by inducing either volume depletion or acute experimental glomerulonephritis (nephrotoxic serum nephritis) all had normal urine RTE concentrations. These results suggest that RTE quantitation by this technique may provide a specific and early quantitative index of the severity of acute nephrotoxic and ischemic renal injury.