Effect of glomerular basement membrane on the initiation of chemiluminescence and lysosomal enzyme release in human polymorphonuclear leucocytes: an in vitro model of glomerular disease.

The effect of glomerular basement membrane (GBM) on the stimulation of human polymorphonuclear leucocytes (PMNL) was investigated using the technique of chemiluminescence (CL). GBM, in the absence of serum, did not initiate CL of PMNL and neither did GBM preincubated with fresh human or rabbit serum. In contrast, GBM preincubated with rabbit anti-human GBM serum (anti-GBM), and then thoroughly washed, initiated an immediate and marked CL response. Heat inactivation of the anti-GBM only partially reduced the CL, suggesting that both Fc and C3b receptors on the PMNL were involved. The addition of fresh, but not heat-inactivated, human serum to PMNL incubated with untreated GBM resulted in a CL response after a short lag. Subsequent analysis of the serum by electrophoresis and immunofixation indicated that the GBM had activated complement. GBM preincubated with anti-GBM was found to elicit the selective release of lysosomal enzymes from PMNL, one of these enzymes degrading the collagen moiety of the GBM. Preincubation of GBM with the serum from a patient with Goodpasture's syndrome also initiated a CL response and a selective release of lysosomal enzymes from PMNL. These results provide additional evidence for imputing a role for PMNL in the pathogenesis of glomerulonephritis. Furthermore, the technique offers a useful in vitro model for the study of immunologically mediated glomerular disease.