Ritchie B W, Niagro F D, Latimer K S, Steffens W L, Pesti D, Aron L, Lukert P D
Department of Small Animal Medicine, College of Veterinary Medicine, University of Georgia, Athens 30602.
J Vet Diagn Invest. 1992 Jan;4(1):13-8. doi: 10.1177/104063879200400104.
Monoclonal antibodies specific for the virus that causes psittacine beak and feather disease (PBFD) were produced by fusing spleen cells from mice immunized with purified concentrated PBFD virus with mouse myeloma cell line Sp2/0. The resulting hybridomas were tested for reactivity against whole purified virus by an enzyme-linked immunosorbent assay (ELISA) system. Four clones, designated 15H8, 8E3, 11G12, and 2C3, were subcloned by limiting dilution. Isotyping indicated that clone 15H8 was secreting IgG, whereas the remaining clones secreted IgM. The secreted immunoglobulins were characterized by reactivity against purified PBFD virus using immunoblotting procedures, by immunohistochemical staining of virus-induced lesions in infected tissues, and by inhibition of PBFD virus agglutination of cockatoo erythrocytes. Antibodies secreted by clones 15H8 and 8E3 had the strongest activity against purified whole virus. Only immunoglobulin secreted by the clone 15H8 could be used to detect viral antigen in infected tissues. None of the monoclonal antibodies had hemagglutination-inhibition activity.
通过将用纯化浓缩的鹦鹉喙羽病（PBFD）病毒免疫的小鼠脾细胞与小鼠骨髓瘤细胞系Sp2/0融合，制备了针对引起鹦鹉喙羽病（PBFD）的病毒的单克隆抗体。通过酶联免疫吸附测定（ELISA）系统检测所得杂交瘤对全纯化病毒的反应性。通过有限稀释法对四个克隆（命名为15H8、8E3、11G12和2C3）进行亚克隆。同型分型表明克隆15H8分泌IgG，而其余克隆分泌IgM。使用免疫印迹程序、通过对感染组织中病毒诱导病变的免疫组织化学染色以及通过抑制PBFD病毒对葵花凤头鹦鹉红细胞的凝集，对分泌的免疫球蛋白进行了针对纯化PBFD病毒的反应性表征。克隆15H8和8E3分泌的抗体对纯化的全病毒具有最强的活性。只有克隆15H8分泌的免疫球蛋白可用于检测感染组织中的病毒抗原。这些单克隆抗体均无血凝抑制活性。
