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首页> 外文期刊>Clinical and diagnostic laboratory immunology >Self-assembly of in vitro-translated human papillomavirus type 16 L1 capsid protein into virus-like particles and antigenic reactivity of the protein.
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Self-assembly of in vitro-translated human papillomavirus type 16 L1 capsid protein into virus-like particles and antigenic reactivity of the protein.

机译：体外翻译的人乳头瘤病毒16型L1衣壳蛋白的自组装成病毒样颗粒和该蛋白的抗原反应性。

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摘要

The human papillomavirus type 16 (HPV-16) L1 capsid protein is the major component of the HPV virion. We prepared L1 protein of HPV-16 in a cell-free system. The L1 gene was cloned in an expression plasmid and transcribed and translated in vitro in a rabbit reticulocyte lysate. The expressed protein had the molecular mass (55 kDa) expected for the L1 protein, and it assembled into virus-like particles that closely resembled papillomavirus virions. The protein retained conformational epitopes, as evidenced by its reactivity with monoclonal antibodies which recognize only intact viral particles. In radioimmunoprecipitation assays with sera from college women grouped by their genital tract HPV DNA status, high reactivity was found in 68% of HPV-16 DNA-positive women, in 23% of women with other HPVs, and in 19% of HPV-negative women. In comparison, none of the sera of children were reactive. The results of the radioimmunoprecipitation assays showed a significant correlation with results obtained with the same sera in an enzyme-linked immunosorbent assay with virus-like particles produced in baculovirus (chi-square test for linear trend, P = 0.0023). Although the amounts of L1 protein obtained are small, the ability to produce virus-like particles by in vitro translation may be useful in the study of virus assembly, virus binding, and the immunological response to HPV infection.
机译：人乳头瘤病毒16型（HPV-16）L1衣壳蛋白是HPV病毒体的主要成分。我们在无细胞系统中制备了HPV-16的L1蛋白。将L1基因克隆到表达质粒中，并在兔网织红细胞裂解物中体外转录和翻译。表达的蛋白质具有预期的L1蛋白质的分子量（55 kDa），并组装成与乳头瘤病毒病毒粒子非常相似的病毒样颗粒。该蛋白保留了构象表位，如其与仅识别完整病毒颗粒的单克隆抗体的反应性所证明。在按女性生殖器HPV DNA状况分组的血清中进行的放射免疫沉淀分析中，发现68％的HPV-16 DNA阳性女性，23％的其他HPV女性以及19％的HPV阴性女性具有高反应性女人。相比之下，儿童的血清均无反应性。放射免疫沉淀试验的结果显示，与杆状病毒中产生的病毒样颗粒的酶联免疫吸附试验中相同血清获得的结果显着相关（线性趋势的卡方检验，P = 0.0023）。尽管获得的L1蛋白量很小，但通过体外翻译产生病毒样颗粒的能力可能对研究病毒装配，病毒结合以及对HPV感染的免疫学反应有用。

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