Ishida S, Feng N, Tang B, Gilbert J M, Greenberg H B
Department of Medicine, Stanford University School of Medicine, California 94305, USA.
J Clin Microbiol. 1996 Jul;34(7):1694-700. doi: 10.1128/JCM.34.7.1694-1700.1996.
The purpose of the present study was to develop a quantitative assay that could be used to measure the local and systemic immune responses to specific rotavirus proteins following rotavirus infection of adult mice. To measure these responses, we used an immunocytochemical staining assay of Spodoptera frugiperda (Sf-9) cells which were infected with recombinant baculovirus expressing selected rotavirus proteins. The specificity of the assay was documented by using a series of monoclonal antibodies to individual rotavirus proteins. We observed that the assay had high levels of sensitivity and specificity for a series of VP7- and VP4-specific neutralizing monoclonal antibodies which recognized conformation-dependent epitopes on their target proteins. We also studied immunoglobulin G (IgG) immune responses in serum and IgA immune responses in the stools of mice infected with wild-type murine rotavirus strain EHPw. In both sera and stools, the most immunogenic proteins were VP6 and VP4. VP2 was less immunogenic than VP6 or VP4, and the immune responses to VP7, NSP2, and NSP4 were very low in serum and undetectable in stools.
本研究的目的是开发一种定量检测方法，用于测量成年小鼠感染轮状病毒后对特定轮状病毒蛋白的局部和全身免疫反应。为了测量这些反应，我们使用了对感染表达选定轮状病毒蛋白的重组杆状病毒的草地贪夜蛾（Sf-9）细胞进行免疫细胞化学染色检测。通过使用针对单个轮状病毒蛋白的一系列单克隆抗体，证明了该检测方法的特异性。我们观察到，对于一系列识别其靶蛋白上构象依赖性表位的VP7和VP4特异性中和单克隆抗体，该检测方法具有高度的敏感性和特异性。我们还研究了感染野生型鼠轮状病毒株EHPw的小鼠血清中的免疫球蛋白G（IgG）免疫反应和粪便中的IgA免疫反应。在血清和粪便中，免疫原性最强的蛋白是VP6和VP4。VP2的免疫原性低于VP6或VP4，并且血清中对VP7、NSP2和NSP4的免疫反应非常低，粪便中无法检测到。
