应用菜粉蝶颗粒体病毒单克隆抗体对不同株病毒进行抗原分析

应用杂交瘤技术,以大菜粉蝶颗粒体病毒(pbGV)、菜粉蝶颗粒体病毒北京分离株(prGV-801)、武汉分离株(prGVw1-78)和济南分离株(prGV-J)等4株病毒为抗原,获得9株杂交瘤细胞(Fr1～9)。ELISA测定细胞培养上清抗体效价最高达8000,腹水效价最高达450000。9株杂交瘤细胞所分泌的抗体,各呈现株或组特异性,其中pr1～4分别与4个毒株起反应,pr5～8可与2～3个毒株起反应,而Pr9则与所有4个毒株均起反应。据此,认为这4个毒株有抗原性差异。不同毒株间抗原性的差异不仅存在于病毒粒子上,而且也存在于颗粒体蛋白上。讨论了昆虫病毒单克隆抗体在鉴别病毒抗原性差异,生物防治和流行病学研究中的意义。

ANTIGENIC ANALYSIS OF VIRUS STRAINS BY MONOCLONAL ANTIBODIES TO PIERIS PAPAE GRANULOSIS VIRUS

Monoclonal antibodies ( McAbs ) to Pieris bressicae granulosis virus (Pb-GV) and three strains of Pieris rapae granulosis viruses, Beijing strain (Pr-GV-801 ) , Wuhan strain ( PrGVWl-78 ) and Jinan strain ( PrGV-Z ) , were generated by polyethylene glycol fusion of splenocytes from immunized Balb/c mice and FO myeloma celis. After extensive screening, 9 hybridoma cell liaes(Prl-9)were selected based on their ability to distinguish the 4 dif ferent strains of GV. The McAbs from these clones (Prl-9)were shown, with ELISA methods, to be directed to 9 different antigenic determinants. McAbs from Prl-4 were reactive specifically to one GV strain. Pr5-8 were directed to epitopes shared by 2-3 GV strains. Pr9 is apparently directed to an antigenic determinant shared by all the strains tested. Differences in an tigenicity is not only expressed in the virions, but also in the granulins.Pr9 has produced the highest titer of supernantants and ascitic fluid. Supernan-tants have ELISA endpoints around 1:8000 and that of ascites around l: 450000.The potential applications of these McAbs" for the classification of GVs, biological control and epidemiological studies are discussed.