流行性出血热病毒单克隆抗体的特性鉴定及其对病毒结构蛋白作用的初步研究

本文运用放射免疫沉淀法(RIP)、Western-blot及ELSA夹心法,对一组针对流行性出血热病毒(EHFV)L99株、C4株的单克隆抗体(McAb)进行特性鉴定,其中4株McAb针对糖蛋白G2,29株针对核壳蛋白(NP),1株针对糖蛋白G1和NP。微量中和试验和血凝抑制(HI)试验分析表明,虽绝大多数抗NP McAb既无中和活性亦无HI活性,但有1株例外。具有明显的中和活性和HI活性,提示EHFV核壳蛋白上可能存在中和抗原和血凝抗原决定簇。2株抗G2 McAb具有较高的HI活性,1株抗G2 McAb有较低的中和活性和较高的HI活性,另一株抗G2 McAb仅具有中和活性,表明EHFV糖蛋白G2上存在独立的中和与血凝抗原决定簇,也可能存在具有中和、血凝双重功能的决定簇。竞争ELISA分析显示。G2蛋白上某些中和位点和血凝位点虽然独立分布,但在某一区域可能相当靠近或有部分重叠。

PRELIMINARY STUDY ON STRUCTURAL PROTEIN OF EPIDEMIC HEMORRHAGIC FEVER VIRUS USING MONOCLONAL ANTIBODIES

A panel of monoclonal antibodies (McAb) against epidemic hemorrha-gic fever virus ( EHFV ) strains L99 and C4 were characterized by Western-blot,radio-immuno-precipitation and sandwich ELISA.Of these McAb, four directed against glycoprotein G2; tweenty-nine against neucleocap-sd protein ( NP ) and one against both NP and glycoprotein Gl of EHFV. The results of neutralization (NT) and hemagglutination inhibition (HI) tests showed that most of the McAbs specific for NP had neither NT nor HI activites, but one had obvious NT and HI activites. It was suggested that there may be some neutralizing and hemagglutinating determinants on NP of EHFV. Two McAb to G2 had only high HI activity; one had both low NT and high HI activites,and another had only NT activity. These data indicate that distinct neutralizing and hemagglutinating epitopes weie located on the G2, and some epitopes had dual-functions of neutralization and hemagglutination.The analysis of competitive ELISA demonstrated that, although some neutralizing and hemagglutinating epitopes were separate, they might be located very close to or overlapping each other.