乙型肝炎病毒感染肾炎患者肾组织病毒抗原和复制中间体的检测及其意义

Detection of HBV antigens and cccDNA in glomeruli of patients with HBV infection and renal disease

目的 :观察乙型肝炎病毒 (HBV)感染者肾组织中三种病毒抗原成分的分布特点及其与HBV感染状态和临床病理之间的联系 ,探讨在肾组织局部是否存在HBV的复制。 方法 :免疫组化法检测合并HBV感染的30例膜性肾病和 1 2例膜增生性肾炎病例的肾活检组织切片中的HBsAg、HBcAg和HBeAg ,同时检测肾小球和循环中的HBV基因组DNA及其复制中间体———闭合环状双链DNA(cccDNA)。 结果 :膜性肾病肾组织中病毒抗原的检出率 (83 3%)显著高于膜增生性肾炎 (33%) ;膜性肾病肾组织检出的抗原以HBcAg和HBeAg多见 ,其中 ,血清HBeAg阳性病例肾组织HBeAg的检出率显著高于HBeAg阴性的病例。膜增生性肾炎肾组织检出的抗原主要是HBeAg。肾组织HBeAg的检出与循环中HBeAg的存在明显相关。伴血清转氨酶升高者肾组织HBV抗原的检出率较转氨酶正常者有升高的趋势。肾小球HBVDNA和cccDNA的检出均与循环中的检测结果高度一致 ,并以伴活动性HBV感染者检出率为高。 结论 :在合并HBV感染的肾炎患者中 ,肾组织HBV抗原的检出率在膜性肾病患者明显高于膜增生性肾炎。肾小球中检出的HBV抗原成分以HBeAg和HBcAg最多见 ,肾小球HBeAg的检出与血清中是否存在HBeAg明显相关。合并肝功能损害者肾组织HBV抗原的检出率较肝功能正常者有增高趋势。

Objective:To investigate the pathogenesis of HBV related glomerulonephritis(HBVGN), the deposition of HBV related antigens in glomeruli were examined, and the association between antigen deposition and state of HBV infection as well as the clinicopathoplocial manifestations were investigated. And in this study, the active replications of HBV in kidney were also examined. Methodology:42 renal patients with HBV infection were involved in this study, including 30 patients whose renal biopsy revealed membranous changes and 12 membranoproliferative changes. Glomerular depositions of HBsAg, HBcAg and HBeAg were detected with immunohistochemistry. PCR and nested PCR were applied in the detection of HBV DNA and cccDNA in isolated glomeruli microscopically dissected from renal biopsy slides, and in the detection of HBV DNA and cccDNA in circulation(including serum and peripheral blood mononuclear cells). The association between clinical manifestations and pathological changes were investigated retrospectively. Results:HBV antigens were detected positive in glomeruli in 83.3% of the patients with membranous changes, and in 33% of the patients with membranous proliferative changes (83.3% vs 33%, P=0.007). In patients with membranous changes, the prodominant pattern of antigens deposition in glomeruli was HBcAg and HBeAg. A significant correlation was found between HBeAg deposition in glomeruli and positivity of HBeAg antigen in serum(P=0.015). Glomerular antigen deposition was more frequently found in patients with elevated serum transaminase. HBV DNA and cccDNA in glomeruli was more frequently found in patients with active HBV infection. Positivity of HBV DNA and cccDNA detection in glomeruli was consistent with the positivity of HBV DNA and cccDNA detection in circulation in most of the patients. Conclusion:The positive rate of HBV antigens in glomeruli in patients with membranous changes was significantly higher than in patients with membranous proliferative changes. HBeAg and HBcAg are main antigens deposited in glomeruli of HBVGN. Significant correlation was found between glomerular HBeAg deposition and HBeAg seropositivity. HBV antigens deposition in gloemruli was more frequently found in patients with liver dysfunction. Consistent findings of cccDNA in glomeruli and circulation make it hard to rule out the influence of circulating cells to the detection of cccDNA in glomeruli.