Hosie M J, Jarrett O
Department of Veterinary Pathology, University of Glasgow, UK.
AIDS. 1990 Mar;4(3):215-20. doi: 10.1097/00002030-199003000-00006.
The proteins of feline immunodeficiency virus (FIV) were identified by sodium dodecylsulphate poly-acrylamide gel electrophoresis (SDS-PAGE) and immunoblotting. Purified [35S]methionine/cysteine-labelled virus contained proteins of Mr 120, 24, 17, and 10kD, of which the most prominent were p24 and p17, and minor components of 62, 54, 52, 41 and 32kD. Sera from FIV-infected cats precipitated two glycoproteins (gp) of Mr 120kD (gp120) and 41kD (gp41) from lysates of [14C]glucosamine-labelled infected cells. Purified virus contained very little or no detectable glycoproteins. The serological response to individual viral proteins was followed in experimentally infected cats by immunoblotting. Since purified virus was a poor source of gp120, a method using FIV-infected cell lysates was developed. Cats produced antibodies to gp120, p55, p24 and p17. (The p55 was presumed to be a precursor of p24 and p17.) Following infection, antibodies developed first to p24 and subsequently to p17, p55 and gp120. Sera from cats infected with three separate isolates of FIV, two from the UK and one from the USA, had cross-reacting antibodies to all of these viral proteins. The criteria for identification of seropositive cats were defined. The minimum requirement for a positive immunoblot was antibody to gp120 or to at least three core proteins (p55, p24 and p17). Comparison of two commercial enzyme-linked immunosorbent assay (ELISA) kits and immunoblotting indicated that false-positive results occurred as a result of non-specific reactions in the ELISA systems.
通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)和免疫印迹法鉴定了猫免疫缺陷病毒(FIV)的蛋白质。纯化的[35S]甲硫氨酸/半胱氨酸标记病毒含有分子量为120、24、17和10kD的蛋白质,其中最主要的是p24和p17,还有分子量为62、54、52、41和32kD的次要成分。来自FIV感染猫的血清从[14C]葡糖胺标记的感染细胞裂解物中沉淀出两种糖蛋白(gp),分子量分别为120kD(gp120)和41kD(gp41)。纯化病毒中几乎没有或检测不到糖蛋白。通过免疫印迹法跟踪实验感染猫对单个病毒蛋白的血清学反应。由于纯化病毒是gp120的不良来源,因此开发了一种使用FIV感染细胞裂解物的方法。猫产生了针对gp120、p55、p24和p17的抗体。(推测p55是p24和p17的前体。)感染后,首先产生针对p24的抗体,随后产生针对p17、p55和gp120的抗体。来自感染了三种不同FIV分离株的猫的血清,其中两种来自英国,一种来自美国,对所有这些病毒蛋白都有交叉反应抗体。定义了鉴定血清阳性猫的标准。免疫印迹阳性的最低要求是针对gp120或至少三种核心蛋白(p55、p24和p17)的抗体。两种商业酶联免疫吸附测定(ELISA)试剂盒与免疫印迹法的比较表明,ELISA系统中的非特异性反应导致了假阳性结果。