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目的分析以MF59为佐剂的脊髓灰质炎灭活疫苗(IPV)免疫恒河猴后诱导的细胞免疫应答。方法将恒河猴分为4组:无佐剂低剂量组(IPVL-PBS组)、低剂量MF59佐剂组(IPVL-MF59组)、无佐剂高剂量组(IPVH-PBS组)和低剂量铝佐剂组(IPVL-AL组)。高剂量组IPV每剂中Ⅰ、Ⅱ、Ⅲ型抗原含量分别为30、32、42 DU,低剂量组IPV每剂中Ⅰ、Ⅱ、Ⅲ型抗原含量分别为3、3.2、4.2 DU,0.5 ml/剂,其中MF59为0.25 ml/剂,氢氧化铝为0.5 mg/剂,均于第1、28、56天经恒河猴后腿肌肉注射,分别于免疫后第28、56、84天采血,分离血清,微量中和试验法检测血清中和抗体效价;分别于免疫后第56和84天,取外周血,分离淋巴细胞,上流式细胞仪检测特异性抗原刺激下的Th1型细胞因子(IL-2、IFNγ、TNF)和Th2型细胞因子(IL-4、IL-5、IL-6)分泌水平,以及CD3+、CD4+、CD8+T淋巴细胞比例的分布情况。结果恒河猴经3次免疫后,IPVL-MF59组血清中和抗体阳转率明显高于其他各组,达75%以上。恒河猴免疫后第56天,IPVL-MF59组IFNγ和TNF分泌水平明显高于IPVL-AL组(P均<0.05),IL-6分泌水平明显高于IPVL-PBS和IPVH-PBS组(P均<0.05);免疫后第84天,各组IL-2、IFNγ、TNF均稳定在同一水平,IL-4、IL-5分泌水平均明显高于第56天(P均<0.05)。免疫后第84天,IPVL-MF59组CD8+T淋巴细胞百分比均明显高于IPVH-PBS和IPVL-PBS组(P均<0.05)。结论 MF59佐剂能增强IPV诱导的TNF和IL-6的分泌水平,增强Th1和Th2类反应。
OBJECTIVE: To analyze the cellular immune response induced by immunization with Rhesus macaque inactivated by MF59 adjuvant poliovirus (IPV). Methods Rhesus macaques were divided into four groups: low dose group (IPVL-PBS group), low dose MF59 group (IPVL-MF59 group), high dose group without adjuvant (IPVH-PBS group) Dose aluminum adjuvant group (IPVL-AL group). In the high-dose IPV group, the content of type I, type II and type III antigens were 30, 32 and 42 DU respectively. The content of type I, type II and type III antigens of IPV in low dose group were 3,3.2,4.2 DU and 0.5 ml / Agent, which MF59 0.25 ml / dose, aluminum hydroxide 0.5 mg / dose, were in the first 1,28,56 days after Rhesus hind muscle injection, respectively, after immunization 28,56,84 days blood , Serum was separated and the serum neutralizing antibody titers were measured by the method of micro-neutralization test. Peripheral blood was collected on day 56 and 84 after immunization, and lymphocytes were isolated. Th1 cytokines stimulated by specific antigens were detected by flow cytometry (IL-2, IFNγ, TNF) and Th2-type cytokines (IL-4, IL-5 and IL-6) as well as the distribution of CD3 +, CD4 + and CD8 + T lymphocytes. Results After 3 immunizations of rhesus monkeys, the positive rate of serum neutralizing antibody in IPVL-MF59 group was significantly higher than that in other groups, reaching over 75%. On the 56th day after immunization, the secretion of IFNγ and TNF in IPVL-MF59 group was significantly higher than that in IPVL-AL group (P <0.05), and the secretion of IL-6 was significantly higher than that in IPVL-PBS and IPVH-PBS group (All P <0.05). On the 84th day after immunization, the levels of IL-2, IFNγ and TNF in each group were all kept at the same level, and the secretion levels of IL-4 and IL-5 were significantly higher than those on the 56th day (all P <0.05). On the 84th day after immunization, the percentage of CD8 + T lymphocytes in IPVL-MF59 group was significantly higher than that in IPVH-PBS and IPVL-PBS groups (all P <0.05). Conclusion MF59 adjuvant can enhance the secretion of TNF and IL-6 induced by IPV and enhance the Th1 and Th2 responses.