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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.

机译:肾小球基底膜对人多形核白细胞化学发光和溶酶体酶释放的影响:肾小球疾病的体外模型。

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摘要

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.
机译:使用化学发光技术(CL)研究了肾小球基底膜(GBM)对人多形核白细胞(PMNL)的刺激作用。在无血清的情况下,GBM不会引发PMNL的CL,GBM也不与新鲜的人或兔血清一起预孵育。相反,GBM与兔抗人GBM血清(anti-GBM)预孵育,然后彻底洗涤,立即引发明显的CL反应。抗GBM的热失活仅部分降低CL,表明PMNL上的Fc和C3b受体均参与。向未处理的GBM孵育的PMNL中添加新鲜但未热灭活的人血清会导致短暂的滞后CL反应。随后通过电泳和免疫固定分析血清表明GBM已激活补体。发现与抗-GBM预温育的GBM引发了从PMNL选择性释放溶酶体酶,这些酶是降解GBM胶原部分的酶之一。 GBM与Goodpasture综合征患者的血清一起预温育还引发CL反应和PMNL中溶酶体酶的选择性释放。这些结果为推断PMNL在肾小球肾炎发病机理中的作用提供了额外的证据。此外,该技术为研究免疫介导的肾小球疾病提供了有用的体外模型。

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