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目的评价CA16 K168/8疫苗候选株免疫血清对不同CA16毒株的交叉中和能力及对致乳鼠麻痹CA16临床分离株的体内中和保护能力。方法将CA16 K168/8疫苗候选株病毒纯化液辅以弗氏佐剂经背部及侧腹部皮下多点注射免疫新西兰大白兔,制备高效价免疫血清。采用微量细胞病变法检测中和抗体效价;用中和抗体效价终浓度为32 U的特异性免疫血清对18株不同CA16临床分离毒株及15株EV71毒株进行交叉中和指数检测;选取中和抗体效价为16、32、64、128、256 U的特异性免疫血清对可致乳鼠产生明显麻痹作用的CA16 FY-18株进行体内中和保护水平检测。结果获得的兔抗CA16 K168/8血清对14株CA16毒株的中和效价为1∶1 024~1∶6 144,GMT值为1∶3 153;可于体外完全中和18株不同CA16毒株,中和指数为100~5 623.4,平均值为794.1,而对15株EV71临床分离株中和指数仅为0.32~5.62,平均值为2.2;体内中和保护水平随抗体效价升高而增加,呈量-效关系,当中和抗体效价达128 U时,可完全保护乳鼠不发生临床麻痹反应。结论 CA16 K168/8疫苗候选株特异性免疫血清体内、外试验结果表明其具有良好的免疫保护性。
Objective To evaluate the cross-neutralization ability of CA16 K168 / 8 vaccine serum against different CA16 strains and the in vivo protection of the CA16 isolated clinical isolate. Methods New Zealand white rabbits were immunized with Freund ’s adjuvant by injecting subcutaneously in the back of the CA16 K168 / 8 vaccine strain and the flanks of the vaccine. The high titer immune serum was prepared. The titer of neutralizing antibody was measured by the method of microcytopathology. The cross-neutralization index of 18 different CA16 clinical isolates and 15 EV71 strains was detected by using specific immune serum with the final antibody titer of 32 U; The neutralizing antibody titers of 16, 32, 64, 128, 256 U were used to detect the level of in vivo neutralization of CA16 FY-18 strain which could cause obvious paralysis in neonatal rats. Results The rabbit anti-CA16 K168 / 8 serum had neutralization titer of 1:1 024 ~ 1: 1444 and GMT value of 1: 153 for 14 CA16 strains. 18 different CA16s could be completely neutralized in vitro The neutralization index was 100-562.34 with an average value of 794.1, while the neutralization index of 15 EV71 clinical isolates was only 0.32-5.62 with an average value of 2.2. The neutralization protection level increased with the antibody titer But increased in a dose-effect relationship. When the neutralizing antibody titer reached 128 U, the clinical paralysis reaction could be completely prevented. Conclusion The results of in vitro and in vivo tests of specific immune serum of CA16 K168 / 8 vaccine candidate showed that it has good immunoprotective properties.