应用单克隆抗体免疫细胞化学法分析尸检组织中肾综合征出血热病毒G2,NP及HA结构蛋白抗原

The Viral Glycoprotein Ⅱ(G2),Nucleoprotein(NP)and Hemagglutinin(HA)Antigens in the Tissues of Cadavers due to the Death of Hemorrhagic Fever with Renal Syndrome Analyzed by Monoclonal Antibodies with Immunocytochemical Methods

本文应用15株分别抗肾综合征出血热(HFRS)病毒糖蛋白(Glycoprotein Ⅱ G2),核蛋白(Nuclcocapsid,NP)及血凝素(Hemagglutinin,HA)抗原的单克隆抗体免疫细胞化学方法对19例HFRS尸检病例的16种组织中的病毒抗原进行了定位和分布的研究及抗原分析。结果表明,死于早期HFRS人体组织内病毒抗原量大,分布广泛,主要以可溶性和颗粒性两种形式存在,前者存在于细胞内和细胞外,呈G2,NP及HA抗原阳性,是参与形成免疫复合物的主要抗原形式;后者是以单纯病毒NP或HA抗原阳性的病毒包涵体(IB)形式存在于细胞内,是病毒直接作用所致细胞病变的标志,IB的广泛分布但只有极个别阳性细胞发生坏死,说明该病毒具有泛嗜性感染和弱致细胞病变能力的特性。抗原分析结果显示,组织细胞中病毒抗原的表达及其抗原量受宿主细胞的种属,组织结构特点及病期的影响,也存在个体差异以及因感染病毒株或血清型不同产生差异的可能性。病毒抗原染色形态学证实,NP上具有HA抗原位点,其抗原决定簇有三类,其中的某些抗原决定簇可因病毒宿主动物或细胞的种属不同,表达也不同。HA抗原在人体组织细胞中的高表达和广泛分布也说明HFRSV对人体有很强的侵袭力。

The viral antigens in 16 kinds of 10% formalin-fixed paraffin-embedded tissues of 19 fatal cases due to the death of hemorrhagic fever with renal syndrome (HFRS) were detected and anslyzed by 15 clones of monoclonal antibodies (MAbs) respectively against glycoprotein Ⅱ(G2), nucleocapsid (NP) and hemagglutinin (HA) antigens of HFRSV with the repeated PAP and repeated PAP com bined with ABC immunocytochemical methods developed in our Lab. The viral antigens, the soluble and granular, were demonstrated to be in large quantities with wide distribution in the tissues of the fatal cases died in the early phase and in less quantities with narrow distribution in the tissue of the fatal cas- es died in the late phases of the disease. The soluble antigens with G2, NP or HA positive,distributed in the cytoplasma and extracellular spaces, were determined to be the components of the soluble im- munocomplexes. The granular antigens,in the forms of inclusion bodies (IB) with NP or HA antigen positive only, localized in the cytoplasma, were the signs of pathologic lesions of the infected cells caused by virus infection. The wide distribution of IB in the bodies with few positive cells necrosis indi- cated that the viruis infection has no porticular predilection for the given host cells,and only mild cy- topathogenic effects on the host cells. The viral antigen analysis showed that three classes of HA anti- genic determinants were determined to be on the NP by the MAbs against HA and morphological evi- dences. The expression and quantities of the viral antigens including some HA antigenic determinants on the NP in the bodies were affected by the species of the host cells, histological features of the tissures and the phases of the disease;the individuality as well as the strains and serotypes of the infected virus difference may also be the influential factors. The wide distribution and high expression of HA in the human tissures also suggested that the virus has the ability of wide infection of the cells in the human bodies.