摘要
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 cellullar immune response induced by inactivated poliovirus vaccine（IPV）containing MF59 adjuvant in rhesus. Methods Rhesus monkeys were divided into IPVL-PBS,IPVL-MF59,IPVH-PBS and IPVL-AL groups,and immunized with adjuvant-free IPV at a low dosage,MF59 adjuvant-containing IPV at a low dosage,adjuvant-free IPV at a high dosage and aluminum adjuvant-containing IPV at a low dosage respectively. The contents of poliovirus antigen of types Ⅰ,Ⅱ and Ⅲ in IPV at a high dosage were 30,32 and 42 DU,while those in IPV at a low dosage were 3,3. 2 and 4. 2 DU,respectively. Each monkey was immunized i.m. with 0. 5 ml of IPV containing 0. 25 ml MF59 or 0. 5 mg aluminum hydroxide on days 1,28 and 56,of which serum samples were collected on days 28,56 and 84 and determined for neutralizing antibody titer by microneutralization test. Peripheral blood was collected on days 58 and 84,from which lymphocytes were isolated and determined for secretion levels of Th1（IL-2,IFNγ and TNF） and Th2（IL-4,IL-5 and IL-6） cytokines stimulated with specific antigen as well as percentages of CD3＋,CD4＋ and CD8＋ T lymphocytes by flow cytometry. Results The serum neutralizing antibody positive rate in IPVLMF59 group after immunization for 3 times was significantly higher than those in the other groups,which was more than75%. The secretion levels of IFNγ and TNF in IPVL-MF59 group on day 56 were significantly higher than those in IPVLAL group（each P〈0. 05）,while that of IL-6 was significantly higher than those in IPVL-PBS and IPVH-PBS groups（each P〈0. 05）. However,on day 84,the IL-2,IFNγ and TNF in various groups were at the same levels,while the IL-4 and IL-5 levels were significantly higher than those on day 56（each P〈0. 05）. The percentage of CD8 ＋ T lymphocytes in IPVL-MF59 group on day 84 was significantly higher than those in IPVH-PBS and IPVL-PBS groups（each P〈0. 05）. Conclusion MF59 adjuvant increased the secretions of TNF and IL-6 induced by IPV and enhanced the Th1
出处
《中国生物制品学杂志》
CAS
CSCD
2014年第10期1263-1267,共5页
Chinese Journal of Biologicals
