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

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7d、21d、28d以及加强免疫7d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28d，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、l：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7d的抗体阳转率达100％；明显高于初次免疫28d和21d；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7d后的白细胞介素100L-10）和肿瘤坏死因子α（TNF-α）的含量与初次免疫7d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。

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 wity 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）-α 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.