Lizano M, López S, Arias C F
Departamento de Biología Molecular, Universidad Nacional autónoma de México, Cuernavaca, Morelos.
J Virol. 1991 Mar;65(3):1383-91. doi: 10.1128/JVI.65.3.1383-1391.1991.
We have previously reported the synthesis in Escherichia coli of polypeptide MS2-VP8', which contains the amino-terminal half of the SA114fM VP4 protein fused to MS2 bacteriophage polymerase sequences (C. F. Arias, M. Lizano, and S. López, J. Gen. Virol. 68:633-642, 1987). In this work we have synthesized the carboxy-terminal half of the VP4 protein also fused to the MS2 polymerase. This protein, designated MS2-VP5', was recognized by sera to the complete virion and was able to induce antibodies to the virus when administered to mice; however, these antibodies had no neutralizing activity. The two chimeric polypeptides were tested for their ability to agglutinate erythrocytes and to prime the immune system of mice. Bacterial lysates enriched for the MS2-VP8' hybrid polypeptide, but not those enriched for the MS2-VP5' protein or those containing proteins from the host E. coli strain, had hemagglutinating activity. This hemagglutination was inhibited by sera to SA114fM rotavirus. In addition, a single dose of the MS2-VP8' polypeptide was able to prime the immune system of mice for an augmented neutralizing antibody response when the animals were subsequently immunized with purified SA114fM virus.
我们之前报道过在大肠杆菌中合成多肽MS2-VP8'，它包含与MS2噬菌体聚合酶序列融合的SA114fM VP4蛋白的氨基末端一半（C.F.阿里亚斯、M.利萨诺和S.洛佩斯，《普通病毒学杂志》68:633-642，1987年）。在这项工作中，我们还合成了同样与MS2聚合酶融合的VP4蛋白的羧基末端一半。这种蛋白命名为MS2-VP5'，能被针对完整病毒体的血清识别，并且在给小鼠注射时能够诱导产生针对该病毒的抗体；然而，这些抗体没有中和活性。对这两种嵌合多肽进行了凝集红细胞以及启动小鼠免疫系统能力的测试。富含MS2-VP8'杂合多肽的细菌裂解物具有血凝活性，而富含MS2-VP5'蛋白的细菌裂解物或含有宿主大肠杆菌菌株蛋白的细菌裂解物则没有。这种血凝反应被针对SA114fM轮状病毒的血清所抑制。此外，当用纯化的SA114fM病毒随后免疫动物时，单剂量的MS2-VP8'多肽能够启动小鼠免疫系统，使其产生增强的中和抗体反应。
