小鼠在柯萨奇病毒A组16型活病毒免疫原性评价中的应用

doi:10.3969/j.issn.1674-5817.2016.06.001

实验动物与比较医学 ›› 2016, Vol. 36 ›› Issue (6): 409-414.DOI: 10.3969/j.issn.1674-5817.2016.06.001

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小鼠在柯萨奇病毒A组16型活病毒免疫原性评价中的应用

孟华清, 杨婷, 谢天宏, 李华, 岳磊, 宋霞, 张也, 谢忠平

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,昆明 650118

收稿日期:2016-08-23 出版日期:2016-12-25 发布日期:2016-12-25
作者简介:孟华清(1991-), 女, 硕士, 研究方向: 病毒性疫苗研究。E-mail: 754759796@qq.com
基金资助:
云南省重点新产品开发计划(2015BC008)

Immunogenicity of Coxsackievirus Group A Type 16 Strains in Mice

MENG Hua-qing, YANG Ting, XIE Tian-hong, LI Hua, YUE Lei, SONG Xia, ZHANG Ye, XIE Zhong-ping

Institute of Medical Biology, Chinese Academy of Medical Sciences and Peking Union Medical College, Yunnan Research Center of Vaccine Engineering Technology on Severe Infections Diseases, Yunnan Key Laboratory of Vaccine Research and Development on Severe Infections Disease, Kunming 650118, China

Received:2016-08-23 Online:2016-12-25 Published:2016-12-25

摘要/Abstract

摘要： 目的评价小鼠在柯萨奇病毒A组16型(CA16)活病毒感染中免疫原性的应用,为减毒活疫苗研究提供实验数据。方法以不同剂量的CA16活病毒通过皮下、腹腔和肌肉途径接种不同年龄的ICR和BALB/c小鼠,采集初次免疫7 d、21 d、28 d以及加强免疫7 d后的血样; 通过微量细胞病变中和法检测中和抗体效价, 细胞因子ELISA检测试剂盒检测6种细胞因子含量。结果初次免疫后28 d,肌肉、腹腔和皮下途径免疫的小鼠血清抗体阳转率分别为83.33%、40.00%和 0.00%,抗体几何平均效价(GMT)分别为1∶169、1∶32和0; 加强免疫后,肌肉、腹腔和皮下途径免疫的小鼠血清抗体阳转率均达100%,抗体GMT分别为1∶813、1∶609和1∶ 32,肌肉和腹腔途径免疫的小鼠血清抗体GMT平均增长4.8倍和19.0倍; 加强免疫7 d的抗体阳转率达100%; 明显高于初次免疫28 d和21 d; BALB/c小鼠与ICR小鼠相比,前者的中和抗体效价和抗体阳转率明显高于后者, 而小鼠年龄对中和抗体效价和抗体阳转率的影响不显著( P>0.05); 免疫剂量对中和抗体效价和抗体阳转率的影响显著(P <0.05), 高剂量病毒免疫小鼠后中和抗体效价和抗体阳转率明显比低剂量高( P<0.05); 细胞因子检测结果显示加强免疫7 d后的白细胞介素10(IL-10)和肿瘤坏死因子a(TNF-a)的含量与初次免疫7 d的相比显著增加。结论 BALB/c和ICR小鼠可做为CA16活病毒免疫原性评价的动物模型,以4~6周龄的BALB/c小鼠最为敏感。

关键词: 柯萨奇病毒A组16型(CA16), 免疫原性, 细胞因子, 减毒活疫苗

Abstract: Objective To evaluate immunogenicity of Coxsackievirus group A type 16 strains (CA16) in mice and provide experimental data for further research. Methods The female BALB/c and ICR mice ageing 4, 6, 8 weeks were inoculated subcutaneously, intraperitoneally and intramuscularly with two different doses of CA16 live virus. Blood were respectively taken at 7, 21, 28 and 35 days after inoculation and serum neutralizing antibody titer was analyzed by micro-neutralization test in vitro and six cytokines were determined by ELISA. Results Seroconversion rates of the mice inoculated intramuscularly, intraperitoneally and subcutaneously were 83.33%, 40.00% and 0.00% at 28 days after the first inoculation respectively, and antibody geometric mean titers (GMT) were 1∶169, 1∶32 and 0. Seroconversion rates of the mice inoculated intramuscularly, intraperitoneally and subcutaneously were 100% at 7 days after the boost, and GMT were 1∶813 (4.8-fold), 1∶609 (19-fold) and 1∶32; Seroconversion rate was 100%, which was significantly higher than that of 28 and 21 days at the first inoculation. Neutralizing antibody titersand seroconversion rates of the Balb/c were significantly higher than that of ICR, but effect of the age on the neutralizing antibody titers and seroconversion rates was not significant (P>0.05). The dose has significantly effect on the neutralizing antibody titer and seroconversion rates (P<0.05). In high dose group, the neutralizing antibody titer and seroconversion rates was significantly higher than low dose. interleukin(IL)-10 and tumor necrosis factor (TNF)-a have a significant increase at 7 days after the boost, compared to the 7 days after the first inoculation. Conclusion BALB/c and ICR mice can be used as animal models of CA16 live virus immunogenicity evaluation, and the most sensitive models was 4-6 week old BALB/c mice.

Key words: Coxsackievirus group A type 16 strains (CA16), Immunogenicity, Cytokine, Attenuated live vaccine

中图分类号:

R373.2+3Q95-33

孟华清, 杨婷, 谢天宏, 李华, 岳磊, 宋霞, 张也, 谢忠平. 小鼠在柯萨奇病毒A组16型活病毒免疫原性评价中的应用[J]. 实验动物与比较医学, 2016, 36(6): 409-414.

MENG Hua-qing, YANG Ting, XIE Tian-hong, LI Hua, YUE Lei, SONG Xia, ZHANG Ye, XIE Zhong-ping. Immunogenicity of Coxsackievirus Group A Type 16 Strains in Mice[J]. Laboratory Animal and Comparative Medicine, 2016, 36(6): 409-414.

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小鼠在柯萨奇病毒A组16型活病毒免疫原性评价中的应用

Immunogenicity of Coxsackievirus Group A Type 16 Strains in Mice

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