Basic Study
Investigation on the production of anti-neutrophil cytoplasmic antibodies in chronic bronchitis rats
Minghua Fan, Ming Zhang, Ye Liang, Shihong Shao, Peng Zhao, Yu Miao, Guangqun Xing
Published 2019-08-15
Cite as Chin J Nephrol, 2019, 35(8): 603-610. DOI: 10.3760/cma.j.issn.1001-7097.2019.08.007
Abstract
ObjectiveTo investigate the pathogenesis of the production of anti-neutrophil cytoplasmic antibodies (ANCA) in the rat models of chronic bronchitis (CB) with recurrent infections.
MethodsThe CB models were made by double element of smoking and lipopolysaccharide (LPS) stimulation. The rats were divided into four groups, including normal control group (n=5), phorbol-12-myristate-13-acetate (PMA)-treated healthy rats control group (n=5), CB rats group (n=5) and PMA-treated CB rats group (n=6). Renal function of rats was detected. The histopathological lung and kidney tissues were observed by HE staining of paraffin section. Immunological markers, including myeloperoxidase anti-neutrophil cytoplasmic antibodies (MPO-ANCA), proteinase 3 anti-neutrophil cytoplasmic antibodies (PR3-ANCA) and citrullinated histone H3 (CitH3), were measured by enzyme-linked immune-sorbent assay (ELISA) at different time points. Correlation between CitH3 and MPO-ANCA was analyzed by the Spearman rank correlation. NETs components were further detected in lung and kidney tissue by confocal immunofluorescence and colocalization analysis.
Results(1) The serum levels of CitH3 and MPO-ANCA in CB+PMA group showed an increased trend. Compared with those in the normal control group and CB rats group, the serum levels of CitH3 and MPO-ANCA in CB+PMA group increased significantly at the sixth week (both P<0.05). Serum CitH3 levels in rats were positively correlated with serum MPO-ANCA levels (rs=0.490, P=0.024). (2) There were pathological manifestations of CB in the lung tissues of rats in CB group and CB+PMA group, and no obvious abnormalities in the lung tissues of rats in the normal control group and control group. In the rat kidney tissue of CB+PMA group, there were inflammatory cells infiltrated in the glomerular and around the renal tubules, but glomerular necrosis was not found. No obvious abnormalities were observed in the kidney tissues of rats in the normal control group, PMA-treated healthy rats control group and CB group. (3) In the lung and kidney tissues of CB+PMA group NETs could be detected by confocal immunofluorescence analysis.
ConclusionCB rats with the recurrent infections can release large amounts of NETs, in which the exposure of MPO antigen will break the immune tolerance and result in the production of MPO-ANCA.
Key words:
Antibodies, antineutrophil cytoplasmic; Bronchitis, chronic; Extracellular traps; Citrullinated histone H3
Contributor Information
Minghua Fan
Department of Nephrology, the Affiliated Hospital of Qingdao University, Qingdao 266555, China (Fan Minghua is Working at Department of Nephrology, People's Hospital of Rizhao, Rizhao City of Shandong Province 276800, China)
Ming Zhang
Department of Nephrology, the Affiliated Hospital of Qingdao University, Qingdao 266555, China
Ye Liang
Key Laboratory, Department of Urology and Andrology, the Affiliated Hospital of Qingdao University, Qingdao 266555, China
Shihong Shao
Department of Pathology, the Affiliated Hospital of Qingdao University, Qingdao 266555, China
Peng Zhao
Department of Clinical Laboratory, the Affiliated Hospital of Qingdao University, Qingdao 266555, China
Yu Miao
Department of Clinical Laboratory, the Affiliated Hospital of Qingdao University, Qingdao 266555, China
Guangqun Xing
Department of Nephrology, the Affiliated Hospital of Qingdao University, Qingdao 266555, China
