CAJ | 학술논문

目的分析肠道病毒71型(EV71)灭活疫苗(人二倍体细胞)对动物的免疫是否能够导致由抗病毒免疫反应所引起的免疫病理损伤,以确定该实验性疫苗的安全性.方法利用恒河猴婴猴作为实验动物,未经免疫的对照组(n=4)和经EV71灭活疫苗免疫的疫苗免疫组(n=4)同时接受EV71病毒攻击后,以临床观察、外周血和脑脊液的抗体和相关炎性因子监测、动物各组织器官的病理学检查分析为指标,判断动物在免疫反应过程中的病理性炎症反应.结果疫苗免疫组婴猴的特异性中和抗体滴度达到1:32,无论病毒攻击与否,其外周血及脑脊液中的不同炎性因子均无明显变化.直接感染组在病毒攻击后,其外周血中嗜碱性粒细胞相比正常值有4～5倍的一过性上升;病毒攻击后第7天外周血中白细胞介素6(IL-6)和γ干扰素(IFN-γ)出现明显上升,直接感染组(平均数分别为18.5、12.7 pg/ml)均高于疫苗免疫组(10.2、7.6 pg/ml).直接感染组脑脊液中IL-6达到了正常值的4～5倍,7 d时,直接感染组和疫苗免疫组分别为102.0 pg/ml和12.4 pg/ml.病理检查提示,疫苗免疫组所有动物中,中枢神经系统的各个部位及其他组织均未见明显病理改变.但在直接感染组中可见到明显的、由病毒感染所导致的病理损伤,包括神经细胞的退行性变、炎细胞浸润、血管袖套等典型的脑炎病理特征,动物支气管及肺组织也出现明显炎症反应.结论 EV71灭活疫苗可以在恒河猴婴猴体内诱导良好的免疫反应,保护恒河猴婴猴在感染EV71时,不发生严重的炎症反应和组织病理损伤.同时,EV71灭活疫苗具有良好的安全性.
목적 분석장도병독71형(EV71)멸활역묘(인이배체세포)대동물적면역시부능구도치유항병독면역반응소인기적면역병리손상,이학정해실험성역묘적안전성.방법 이용항하후영후작위실험동물,미경면역적대조조(n=4)화경EV71멸활역묘면역적역묘면역조(n=4)동시접수EV71병독공격후,이림상관찰、외주혈화뇌척액적항체화상관염성인자감측、동물각조직기관적병이학검사분석위지표,판단동물재면역반응과정중적병이성염증반응.결과 역묘면역조영후적특이성중화항체적도체도1:32,무론병독공격여부,기외주혈급뇌척액중적불동염성인자균무명현변화.직접감염조재병독공격후,기외주혈중기감성립세포상비정상치유4～5배적일과성상승;병독공격후제7천외주혈중백세포개소6(IL-6)화γ간우소(IFN-γ)출현명현상승,직접감염조(평균수분별위18.5、12.7 pg/ml)균고우역묘면역조(10.2、7.6 pg/ml).직접감염조뇌척액중IL-6체도료정상치적4～5배,7 d시,직접감염조화역묘면역조분별위102.0 pg/ml화12.4 pg/ml.병리검사제시,역묘면역조소유동물중,중추신경계통적각개부위급기타조직균미견명현병리개변.단재직접감염조중가견도명현적、유병독감염소도치적병리손상,포괄신경세포적퇴행성변、염세포침윤、혈관수투등전형적뇌염병리특정,동물지기관급폐조직야출현명현염증반응.결론 EV71멸활역묘가이재항하후영후체내유도량호적면역반응,보호항하후영후재감염EV71시,불발생엄중적염증반응화조직병리손상.동시,EV71멸활역묘구유량호적안전성.
Objective To evaluate the safety of enterovirus type 71 (EV71) inactivated vaccine (human diploid derived) for infection prevention in an animal model by investigating the immune responses and related patho-inflammatory reactions. Methods In the neonatal monkey model for EV71 vaccine protection, vaccinated group (n=4) and unvaccinated group (n=4) were attacked with live virus at the same time, the parameters of clinical observations, antibodies and inflammatory factors in peripheral blood and cerebrospinal fluid (CSF) were detected. And the pathological changes in major organs were used to determine the patho-inflammatory reactions during the immune responses elicited by vaccination. Results The neutralizing antibodies of vaccine group reach to 1∶ 32. There was no obvious changes of inflammatory factors in peripheral blood and CSF of monkeys challenged or unchallenged by live virus. In peripheral blood of unvaccinated group, the level of basophilic granulocyte higher 4-5 times than normal level and the interferon-γ(IFN-γ) showed obvious increase. Live virus infected after 7 days, the interleukin-6(IL-6)and IFN-γ in peripheral blood of unvaccinated group(18.5、12.7 pg/ml)were higher than vaccinated group (10.2、7.6 pg/ml). Furthermore, the IL-6 in CSF(102.0 pg/ml) had 4-5 times increased than vaccinated group (12.4 pg/ml) at 7 days after virus exposure. Meanwhile, the pathological analysis revealed that no obvious changes were detected in CNS and other organs of vaccinated monkeys challenged with live virus. However, the pathological damages induced by virus infection could be determined in the unvaccinated control monkeys, including neuronal damage, massive cellular infiltration associated with pulmonary edema/hemorrhage and pulmonary/bronchial damage due to an infiltration of inflammatory cells. Conclusion Capable of inducing an immune response, the EV71 inactivated vaccine offers protection to neonatal rhesus monkeys against the attacks of live virus. Based on the results of no patho-inflammatory reaction and pathological damage after viral infection in vaccinated animals, the excellent safety of this vaccine may be confirmed in neonatal monkey.