肺炎链球菌溶血素ΔA146Ply黏膜免疫的保护效果  

Protective Effect of Mucosal Immunization with ΔA146Ply against Streptococcus pneumoniae

作　　者：姚润[1] 崔亚利[1] 袁军[1] 王虹[1] 龚艺[1] 尹一兵[1] 张雪梅[1] 

机构地区：[1]重庆医科大学检验系临床检验诊断学教育部重点实验室,重庆400016

出　　处：《中国生物制品学杂志》2011年第5期579-582,共4页Chinese Journal of Biologicals

基　　金：国家自然科学基金(31070819)

摘　　要：目的探讨肺炎链球菌146位氨基酸缺失的溶血素ΔA146Ply蛋白的保护作用,评价其作为肺炎链球菌疫苗候选蛋白的可行性。方法将BALB/c小鼠分为2组:试验组(15μgΔA146Ply蛋白与1 mg/ml CT佐剂的混合物30μl)和对照组(PBS与CT佐剂的混合物30μl),经鼻腔免疫小鼠,每周1次,连续4周。末次免疫1周后,ELISA检测免疫小鼠血清及唾液中特异性抗体水平;并进行ΔA146Ply对多种血清型肺炎链球菌感染的主动保护试验及其特异性抗血清的被动保护试验;West-ern blot分析Ply蛋白在肺炎链球菌中的保守性;间接ELISA检测免疫小鼠血清IgG抗体亚型和细胞因子IL-17A水平。结果试验组小鼠血清中IgG效价为5.12×105,唾液中IgG及IgA效价分别为4.0×103和4.8×103;ΔA146Ply黏膜免疫可有效延长小鼠的生存时间,并提高其生存率;其对肺炎链球菌NCTC7466、CMCC(B)31436、CMCC(B)31614和CMCC(B)31207的保护率分别为50.0%、41.7%、50.0%和41.7%;被动免疫保护试验试验组小鼠的存活率为70%;ΔA146Ply黏膜免疫诱导的抗体亚型主要为IgG1和IgG2b,IL-17A的水平高达(678.55±189)pg/ml,于刺激后48 h达峰值。Ply在4种菌株中均具有良好的保守性。结论ΔA146Ply黏膜免疫可诱导小鼠产生体液免疫和细胞免疫反应,能有效抵抗多株致病性肺炎链球菌的感染,提示ΔA146Ply是肺炎链球菌较好的疫苗候选蛋白。Objective To investigate the protective effect of ΔA146Ply protein（hemolysin of Streptococcus pneumonia with a single deletion of A146） and the feasibility of the said protein as a candidate S.pneumonia vaccine.Methods BALB / c mice were divided into test and control groups,and immunized i.n.with 30 μl of mixture of 15 μg ΔA146Ply with 1 mg / ml CT adjuvant and 30 μl of mixture of PBS with CT adjuvant respectively,once a week for 4 weeks.The specific antibody levels in sera and saliva of mice were determined by ELISA 1 week after the last immunization.Meanwhile,active protection test against S.pneumonia of several serotypes was performed on ΔA146Ply,while passive protection test on antisera of ΔA146Ply.The conservation of Ply protein in S.pneumonia was analyzed by Western blot.The IgG subtype and IL-17A level in sera of mice were determined by indirect ELISA.Results The IgG titer in sera of mice in test group was 5.12 × 105,while IgG and IgA titers in saliva were 4.0 × 103 and 4.8 × 103 respectively.Mucosal immunization with ΔA146Ply prolonged the survival time and increased the survival rate of mice effectively.The protective rates to S.pneumonia NCTC7466,CMCC（B） 31436,CMCC（B） 31614 and CMCC（B） 31207 were 50.0%,41.7%,50.0% and 41.7% respectively.In passive protection test,the survival rate of mice in test group was 70%.The subtypes of antibodies induced by mucosal immunization with ΔA146Ply were mainly IgG1 and IgG2b.IL-17A level reached a peak value of（678.55 ± 189.00） pg / ml 48 h after treatment.Ply was highly conserved in all the 4 S.pneumonia strains.Conclusion Mucosal immunization with ΔA146Ply induced both humoral and cellular immune responses against infections with several S.pneumonia strains effectively,indicating that ΔA146Ply might be a good candidate protein of S.pneumonia vaccine.

关 键 词：链球菌 肺炎 链球菌溶血素类 免疫 黏膜 保护效果 小鼠 

分 类 号：R378.14[医药卫生—病原生物学] R392-33[医药卫生—基础医学]