Zhu X L, Paul P S, Vaughn E, Morales A
Veterinary Medical Research Institute, College of Veterinary Medicine, Iowa State University, Ames 50011.
Am J Vet Res. 1990 Feb;51(2):232-8.
Hybridomas secreting monoclonal (MAB) to transmissible gastroenteritis virus (TGEV) were produced by fusion of SP2/0 myeloma cells and splenic lymphocytes of BALB/c mice immunized with the virulent cell-passaged Miller strain of TGEV. The MAB secreted by these hybridomas were partially characterized; 4 of them (MA4, MA5, MH11, MB2) had high-neutralization titer for TGEV. The remaining 7 (MC6, MD9, ME5, MG5, MF2, ME9, MG7) did not neutralize TGEV at 1:25 dilution. All 4 neutralizing and 2 of the nonneutralizing MAB reacted with the E2 protein of TGEV in a radioimmunoprecipitation assay. The remaining 5 MAB reacted with the E1 protein of TGEV. Reactivity of the MAB was tested in an indirect immunofluorescent assay with 3 cell culture-adapted strains of TGEV (Miller, Purdue, and Illinois) and 13 wild-type isolates of TGEV. Neutralizing MAB reacted with all 13 wild-type isolates and the 3 cell culture-adapted strains of TGEV. In contrast, nonneutralizing MAB that reacted with the Miller strain of TGEV varied in their reactivity with the wild-type TGEV isolates. Reactivity of neutralizing MAB was also tested, using plaque-reduction neutralization assays with Miller, Purdue, and Illinois strains and 5 wild-type isolates. All 4 neutralizing MAB neutralized the 8 virus isolates, but the neutralization titer was higher with the homologous virus than with the heterologous virus isolates. However, neutralization titers of the 4 neutralizing MAB were 4 to 16 times higher for the homologous Miller strain of TGEV than for the heterologous Illinois and Purdue strains, and were 4 to 1,000 times higher than for the wild-type isolates.(ABSTRACT TRUNCATED AT 250 WORDS)
通过将 SP2/0 骨髓瘤细胞与用 TGEV 强毒株细胞传代的 Miller 株免疫的 BALB/c 小鼠的脾淋巴细胞融合,产生了分泌抗传染性胃肠炎病毒(TGEV)单克隆抗体(MAB)的杂交瘤。对这些杂交瘤分泌的单克隆抗体进行了部分特性鉴定;其中 4 种(MA4、MA5、MH11、MB2)对 TGEV 具有高中和效价。其余 7 种(MC6、MD9、ME5、MG5、MF2、ME9、MG7)在 1:25 稀释时不能中和 TGEV。在放射免疫沉淀试验中,所有 4 种中和性单克隆抗体和 2 种非中和性单克隆抗体都与 TGEV 的 E2 蛋白发生反应。其余 5 种单克隆抗体与 TGEV 的 E1 蛋白发生反应。用 3 种适应细胞培养的 TGEV 毒株(Miller、Purdue 和 Illinois)和 13 种 TGEV 野生型分离株,通过间接免疫荧光试验检测了单克隆抗体的反应性。中和性单克隆抗体与所有 13 种野生型分离株以及 3 种适应细胞培养的 TGEV 毒株发生反应。相比之下,与 TGEV 的 Miller 株发生反应的非中和性单克隆抗体与野生型 TGEV 分离株的反应性各不相同。还使用对 Miller、Purdue 和 Illinois 毒株以及 5 种野生型分离株的蚀斑减少中和试验,检测了中和性单克隆抗体的反应性。所有 4 种中和性单克隆抗体都能中和这 8 种病毒分离株,但同源病毒的中和效价比异源病毒分离株更高。然而,4 种中和性单克隆抗体对同源 TGEV 的 Miller 株的中和效价比异源的 Illinois 和 Purdue 株高 4 至 16 倍,比野生型分离株高 4 至 1000 倍。(摘要截短于 250 字)