Monoclonal antibodies to a virulent strain of transmissible gastroenteritis virus: comparison of reactivity with virulent and attenuated virus.
Monoclonal antibodies to a virulent strain of transmissible gastroenteritis virus: comparison of reactivity with virulent and attenuated virus.
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DOI：
10.1007/bf01311003
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发表时间：
1988
影响因子：
2.7
通讯作者：
Saif LJ
中科院分区：
文献类型：
--
作者：
Welch SK;Saif LJ
产生并表征了 12 个分泌针对传染性胃肠炎病毒 (TGEV) 米勒毒株的单克隆抗体 (MAb) 的杂交瘤。在细胞培养免疫荧光 (CCIF) 测定中，三种针对 pplomer 蛋白 (E 2) 的 MAb 具有核周荧光，四种未分类的 MAb 显示细胞膜荧光。这七种单克隆抗体中的六种可中和减毒型和强毒型 TGEV，而第七种（未分类的单克隆抗体）仅可中和后一种病毒。两种能够结合受感染细胞细胞膜的 MAb 具有较低的中和抗体滴度（8 至 72），但能够区分强毒和减毒 TGEV（中和滴度差异为 9 至 72 倍）。两种 E 2 特异性 MAb 具有较高的中和抗体滴度（782 至 34,117），并且针对减毒和强毒 TGEV 株的滴度显示出 4 至 13 倍的差异。五种对核衣壳 (N) 蛋白具有特异性的 MAb 在 CCIF 中具有细胞质颗粒荧光，并且不会中和 TGEV。 MAb 与 TGEV 强毒株和减毒株的 CCIF 抗体滴度比较表明，大多数 E 2 和所有 N 特异性 MAb 的滴度存在差异，针对强毒 TGEV（同源株）的滴度始终较高。在无菌猪中制备的针对减毒、强毒和最近分离的 TGEV 的超免疫抗血清可免疫沉淀减毒和强毒 TGEV 菌株的 3 种主要结构蛋白。相对摩尔数重量。使用超免疫猪血清或单克隆抗体揭示了两种病毒株之间 E 1 和 E 2 蛋白的差异。除了 48 K N 蛋白外，超免疫血清和 MAb 还可以免疫共沉淀 44 K 蛋白，但主要来自减毒 TGEV 的裂解物。
Twelve hybridomas secreting monoclonal antibodies (MAbs) against Miller virulent strain of transmissible gastroenteritis virus (TGEV) were generated and characterized. In a cell culture immunofluorescence (CCIF) assay, three MAbs directed against peplomer protein (E 2) had perinuclear fluorescence and four unclassified MAbs showed cell membrane fluorescence. Six of these seven MAbs neutralized both attenuated and virulent TGEV, and the seventh (an unclassified MAb) neutralized only the latter virus. Two MAbs able to bind the cell membrane of infected cells had low neutralizing antibody titers (8 to 72) but were able to distinguish between virulent and attenuated TGEV (9- to 72-fold differences in neutralizing titers). Two E 2-specific MAbs had higher neutralizing antibody titers (782 to 34,117) and showed 4- to 13-fold differences in titers against the attenuated and virulent TGEV strains. Five MAbs which were specific for nucleocapsid (N) protein had cytoplasmic, particulate fluorescence in CCIF, and did not neutralize TGEV. Comparison of CCIF antibody titers of MAbs to the virulent and attenuated strains of TGEV indicated that differences existed in titers of most E 2 and all N-specific MAbs, with titers consistently higher against virulent TGEV (homologous strain). Hyperimmune antisera prepared in gnotobiotic pigs against the attenuated, virulent and a recent isolate of TGEV immunoprecipitated the 3 major structural proteins of both the attenuated and virulent TGEV strains. Relative mol. wt. differences in the E 1 and E 2 proteins between the two virus strains were revealed using either the hyperimmune pig sera or MAbs. In addition to the 48 K N protein, a 44 K protein was coimmunoprecipitated by the hyperimmune sera and MAbs, but mainly from lysates of attenuated TGEV.