Direct evidence of the production of IgA by tonsillar lymphocytes and the binding of IgA to the glomerular mesangium of IgA nephropathy patients.

Tonsils were obtained by tonsillectomy from a 33-year-old woman with IgA nephropathy. Hetero-hybridoma cells of human tonsillar B-lymphocytes with mouse myeloma cells (NS-1) were made, and cultured in HAT medium supplemented with 10% fetus bovine serum and hybridoma cloning factor. The culture medium was analyzed by Western blot analysis using anti-human IgA antibody, and both IgA1 and IgA2 were demonstrated to be produced. The specimens of the biopsied kidney tissue of IgA nephropathy were washed with 0.02 M citrate buffer (pH 3.2) to remove deposited IgA from glomerulus. The specimens were then incubated with the culture media of hybridoma cells, and immuno-fluorescence analysis using FITC-conjugated anti-human IgA antibody was performed. IgA deposit was efficiently removed by washing with citrate buffer and was recovered after incubation with the culture medium of hybridoma cells. Direct evidence of binding of IgA produced by tonsillar B-lymphocytes to the glomerular mesangium of IgA nephropathy was demonstrated.