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应用原位分子杂交技术和免疫组织化学方法,以鼠抗人的TNF-α单克隆抗体、质粒pUC19含有TNF-αcDNA经异羟基洋地黄苷配基-11-dUTP随机引物标记作为探针,检测17例流行性出血热(EHF)尸检肾组织中TNF-α原位表达。结果显示,EHF尸检肾组织中TNF-α性率为82.35％,TNF-α阳性主要见于近曲小管,其阳性强度与肾小管病变程度相一致;连续性切片显示,4例CIB阳性的集合管上皮细胞同时检测到TNF-α阳性。肾间质的单核细胞表达TNT-αRNA(16/17例),提示EHF肾组织原位产生TNF-α,并参与了肾小管的损伤。
In situ hybridization and immunohistochemistry were performed using mouse anti-human TNF-α monoclonal antibody. Plasmid pUC19 containing TNF-α cDNA was labeled with a random primer labeled with digoxygenin-11-dUTP as a probe to detect TNF-α was expressed in situ in 17 cases of hemorrhagic fever with hemorrhagic fever (EHF) autopsy. The results showed that the positive rate of TNF-α in renal tissue of EHF autopsy was 82.35%. The positive expression of TNF-α was mainly found in proximal convoluted tubules. The positive intensity of tubular necrosis was consistent with the degree of renal tubule lesion. Epithelial cells were also detected TNF-α-positive. TNT-αRNA (16/17 cases) was expressed in renal interstitial mononuclear cells, which suggested that TNF-α was produced in situ in EHF renal tissues and involved in renal tubular injury.