柯萨奇B3病毒单衣壳蛋白或纯化病毒颗粒在小鼠体内诱导抗体的评价

T. Shimoyama, T. Kubota, J. Shirai, R. Watanabe
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引用次数: 0

摘要

研究了柯萨奇病毒B3 (CVB3) 4种衣壳蛋白(VP1、VP2、VP3和VP4)在大肠杆菌表达系统中的重组表达,并将其作为ICR小鼠抗CVB3亚单位疫苗的抗原。抗原表达为硫氧还蛋白-组氨酸(TrxHis)标记蛋白,免疫前纯化。尽管除了VP4之外的所有vp都能在小鼠体内诱导抗CVB3特异性抗体(通过ELISA和western blotting检测),但在病毒中和实验中,它们不能中和感染性CVB3。同时,根据其在HeLa细胞上的斑块大小,从CVB3病毒原液中纯化出2株病毒株。用2种纯化病毒株(s株和l株)和未纯化病毒株(野生型)感染ICR小鼠,分析纯化病毒株和未纯化病毒株感染的抗体应答差异。采用ELISA法检测抗血清对各病毒株的反应性,结果表明,纯化的病毒株接种后可诱导对接种株产生较强的抗体应答。结果,对野生型和其他病毒株的抗体反应被抑制。这些结果表明,使用未纯化的病毒原液作为抗原有利于在接种动物中诱导广泛的抗体反应。
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Evaluation of Antibodies Induced by the Injection of Single Capsid Protein or Purified Virus Particle of Coxsackievirus B3 in Mice
Four capsid proteins (VP1, VP2, VP3, and VP4) of coxsackievirus B3 (CVB3) were expressed as recombinant proteins in an Escherichia coli expression system and used as antigens for subunit vaccines against CVB3 in ICR mice. Antigens were expressed as thioredoxin-histidine (TrxHis)-tagged protein and purified before immunization. Although all VPs other than VP4 induced anti-CVB3 specific antibodies in mice (detected by ELISA and western blotting), they did not neutralize the infectious CVB3 in a virus neutralization assay. Meanwhile, 2 virus strains were purified from CVB3 virus stock on the basis of their plaque size on HeLa cells. ICR mice were infected with the 2 purified virus strains (S-strain and L-strain) and unpurified virus stock (wild type) to analyze the difference in antibody responses against infections of purified and unpurified virus strains. The reactivity of antisera against each virus strain was tested by ELISA, and the results showed that the inoculation of purified virus strain induced a strong antibody response against the inoculated strain. As a result, the antibody response against wild-type and other virus strains was suppressed. These results suggest using unpurified virus stock as an antigen is advantageous for inducing a broad antibody response in inoculated animals.
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