柯萨奇病毒A组16型疫苗候选株减毒特性的分析  

Characteristics of live attenuated coxsackievirus A16 candidate vaccine strain

作　　者：杨水芝[1] 谢天宏[1] 李华[1] 姜广菊 龙润乡[1] 杨婷[1] 岳磊[1] 罗芳宇 朱凡丽 谢忠平[1] 

机构地区：[1]中国医学科学院北京协和医学院医学生物学研究所云南省重大传染病疫苗工程技术研究中心云南省重大传染病疫苗研发重点实验室,云南昆明650118

出　　处：《中国生物制品学杂志》2015年第5期441-448,共8页Chinese Journal of Biologicals

基　　金：云南省重点新产品开发计划项目(2012BC006)

摘　　要：目的对柯萨奇病毒A组16型（coxsackievirus A16,CA16）减毒活疫苗候选株进行初步分析,为CA16减毒活疫苗的研制奠定基础。方法将两株CA16疫苗候选株（KM168和KM154）进行低温连续传代减毒,对不同代次病毒进行减毒特性的分析。将候选株病毒收获液免疫ICR乳鼠（颅内注射）及ICR成鼠（腹腔注射）,分别评价减毒疫苗候选株的毒力、免疫原性以及乳鼠作为减毒活疫苗残余毒力评价动物模型的可行性;对病毒进行VP1片段的测序,分析遗传稳定性。结果两个候选株经低温连续传代后,均具有稳定的生长特性及良好的免疫原性,感染性滴度均保持在约7.0 Lg CCID50/ml,免疫小鼠后抗体阳转率达100%,最高GMT约1∶30。乳鼠及成鼠感染病毒后,在大部分组织中能检测到CA16病毒;KM154株在第12代后未对乳鼠产生临床致病性及致死率,病理检测结果主要表现为少量炎性细胞集结,无明显组织损伤,随着传代增加,组织损伤明显减少。两个候选株经低温连续传代后,能保证VP1片段的遗传稳定性。结论低温连续传代可使CA16的毒力明显降低,有望筛选出减毒活疫苗的候选株;1~2日龄ICR乳鼠可以作为CA16减毒活疫苗的候选株毒力评价的动物模型。Objective To preliminarily analyze the characteristics of live attenuated coxsackievirus A16（CA16） vaccine candidate strain and lay a foundation of development of the vaccine. Methods Two CA16 vaccine candidate strains,KM168 and KM154, were attenuated by continuous subculture at low temperatures, and the characteristics of various passages were analyzed. Suckling and adult ICR mice were immunized with the candidate strain by intracerebral and intraperitoneal injections respectively to evaluate the virulence and immunogenicity as well as the feasibility of newborn mice as a model for evaluation of residual virulence of live attenuated vaccine. The VP1 fragment of virus was sequenced to analyze the genetic stability. Results Both the cold-adapted candidate strains showed stable growth characteristics and good immunogenicity, of which the infectious titers were about 7. 0 Lg CCID50/ ml. The antibody positive conversion rate of immunized mice reached 100%, while the maximum GMT reached 1 ∶ 30. CA16 was detected in most of the tissues of suckling and adult mice. KM154 strain of more than passage 12 showed no pathopoiesis or lethality to suckling mice, of which the pathological examination showed aggregation of a small quantity of inflammatory cells while no obvious injury of tissue, and the injury decreased significantly with the increasing passages. The genetic stability of VP1 fragment was ensured after subculture of the two candidate strains at a low temperature. Conclusion Subculture at a low temperature decreased the virulence of CA16 significantly, indicating that it was hopeful to screen the candidate strains for live attenuated vaccine. Suckling ICR mice at age of 1 ~ 2 d might be used as an animal model for evaluation of virulence of live

关 键 词：柯萨奇病毒 疫苗 减毒株 

分 类 号：R373.23[医药卫生—病原生物学] R392-33[医药卫生—基础医学]