Quantitative studies of tubular immune complex formation and clearance in rats

Ishidate, Takeo; Ward, Harry J.; Hoyer, John R.
December 1990
Kidney International;Dec1990, Vol. 38 Issue 6, p1075
Academic Journal
Tubular antibody deposition and clearance was quantitatively studied using affinity-purified rabbit antibodies to rat Tamm-Horsfall protein (TH), a surface membrane glycoprotein of the tubular cells of the thick ascending limb of the loop of Henle. Immune complexes are formed in situ at the base of these cells in rats injected with antisera to TH. The renal binding of I125-anti-TH was determined in pair label studies. Kidneys and other organs were removed from groups of rats for isotope counting at four hours to 14 days after an injection of I125-anti-TH and l131-normal rabbit IgG. The greatest total renal anti-TH binding after injection of 500 μg of anti-TH was observed at 24 hours in normal rats (18.55 ± 1.6 μg). During the period of most rapid clearance (day 2 to day 7) the half life of renal anti-TH binding (84.2 hours) and the half life of anti-TH in the serum (68.5 hours) were shorter than that of IgG in the serum (117.8 hours). There was no substantial uptake of anti-TH by other organs. A close relationship between serum levels and renal uptake of anti-TH at 24 hours was also observed in rats given from 50 to 6000 μg of anti-TH: renal saturation was evident only at the highest dose. This close relationship was also present during the clearance phase in rats injected with 3700 μg of anti-TH: the half life of anti-TH was 96.2 hours in kidneys and 110 hours in serum while the half life of rabbit IgG in serum was 151.8 hours. Markedly increased renal uptake of anti-TH was observed in proteinuric rats with passive Heymann nephritis. In very proteinuric rats, 14.1% of the injected dose was bound to kidneys at 24 hours, in these rats, serum anti-TH levels decreased very rapidly to 4% of control serum levels by five days. Throughout the period of study, the serum levels of anti-TH determined by direct radiometric assay corresponded very closely to those obtained by enzyme-linked immunosorbent assay (ELISA). Urinary excretion was a major mechanism for the clearance of anti-TH in proteinuric rats: more than 10% of the injected I123- anti-TH was recovered intact (that is, protein bound) during the first day after injection. During the clearance phase for renal deposits, urinary clearance of anti-TH exceeded urinary clearance of IgG due to release of renal bound antibody into urine. These quantitative studies have defined a new pattern of renal antibody binding kinetics in normal rats which differs from patterns observed with intrinsic glomerular antigens; a close relationship between serum levels and renal binding of antibody over a broad dosage range was apparent during both uptake and clearance phases. They also show that TH is present on cell surfaces only in the kidney, and that increased glomerular permeability greatly alters the kinetics of tubular antibody deposition and clearance.


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