A SIMPLE METHOD TO IDENTIFY NBT-POSITIVE CELLS IN ISOLATED GLOMERULI

Background, Reactive oxygen radicals are probably involved in the path ogenesis of human and experimental models of renal disease, yet curren t methods are inadequate to quantify and identify the cells producing reactive oxygen radicals. Methods and results. We used the nitroblue t etrazolium reaction to determine superoxide anion production in glomer ular cells in phorbol myristate-stimulated glomerular suspensions and in isolated glomeruli from rats with nephrotoxic nephritis, ureteral o bstruction, and puromycin aminonucleoside nephrosis. We were also able to identify these nitroblue tetrazolium + cells using specific approp iate antibodies. When the technique was tested in conditions known to increase reactive oxygen radicals, as phorbol myristate-stimulated glo meruli and glomeruli from animals with nephrotoxic nephritis and urete ral obstruction, increased number of nitroblue tetrazolium + cells wer e found. These cells were identified as glomerular intrinsic cells (Th y-1+) or infiltrating; leukocytes (leukocyte common antigen+ or antine utrophil+). Conclusion. This method may be useful to determine cells p articipating in glomerular damage induced by reactive oxygen radicals.