细胞因子佐剂白介素23和佐剂CpG2216对呼吸道合胞病毒重组疫苗免疫原性的影响  

Effect of IL-23 and CpG2216 on the immunogenity of the respiratory syncytial virus recombinant protein vaccine

作　　者：张慧贤[1] 魏林[2] 李彩霞[2] 刘建勋[2] 崔玉秀[2] 海燕[2] 刘颖[2] 曾瑞红[2] 

机构地区：[1]河北省血液中心,石家庄050061 [2]河北医科大学免疫教研室,石家庄050017

出　　处：《国际呼吸杂志》2011年第11期853-857,共5页International Journal of Respiration

基　　金：基金项目：国家自然科学基金（30972801）;教育部科技重点项目（209009）和教育部高等学校博士学科专项科研基金（20091323120005）

摘　　要：目的研究细胞因子佐剂白介素23（IL-23）和佐剂CpG2216。对呼吸道合胞病毒（RSV）重组疫苗G1F／M2免疫原性的影响。方法将编码IL-23的质粒（简称IL-23）单独作为佐剂以及将IL-23和CpG2216联合作为佐剂与GIF／M2共免疫BABL／c小鼠三次，在每次免疫后10d取血，分离血清，用间接ELISA法测定三次免疫后的血清IgG抗体效价，观察佐剂对体液免疫的影响。第3次免疫后2周处死小鼠，取脾细胞，用LDH法检测CTI。活性、ELISPOT法检测分泌IL－4和7－干扰素（IFN－7）的淋巴细胞水平、流式细胞仪检测CD4＾＋、CD8＾＋的效应记忆和中央记忆T细胞，观察佐剂对细胞免疫的影响。结果ELISA结果表明：除PBS组外，各佐剂组均诱导了高滴度的IgG抗体，其中IL－23＋G1F／M2组的IgG水平显著低于PBS＋G1F／M2组和IL23＋CpG2216＋GIF／M2组，另外，PBS＋GIF／M2组与IL－23＋CpG2216相比差异无统计学意义；CTL杀伤实验显示：IL－23＋CpG2216＋G1F／M2组的CTL杀伤活性显著高于其他组；ELISPOT结果显示IL－23＋CpG2216＋G1F／M2组分泌IFN一7淋巴细胞数量显著多于其他组，且各组分泌IFN－γ的淋巴细胞数显著多于分泌IL－4的淋巴细胞；流式细胞仪检测结果表明，IL－23和IL23＋CpG2216对G1F／M2诱导产生记忆细胞有明显增强作用。结论佐剂IL－3和佐剂CpG2216联合使用能够增强RSV重组疫苗的免疫应答。Objective To investigate the role of IL-23 and CPG2216 adjuvant in the cellular immune response induced by recombinant protein vaccine of respiratory syncytial virus （RSV）. Methods BABL/c mice were immunized with RSV recombinant vaccines GIF/M2 protein with pcDNA3.1 IL -23 eukaryotic expression plasmid（IL-23） combined or alone with CpG2216 as immunologic adjuvant. An interval of 10 days was invariably employed for second, and third immunizations. Mice were phlebotomized 10 days after every immunization and the serum was separated. Specific antibody IgG titers in sera from vaccinated mice were assessed by indirect ELISA. Spleens from the mice immunized were removed two weeks after the third immunization. Specific lysis activity was assessed using a LDH Cytotoxicity Assay Kit. IFN-7- and IL-4-secreting cells were quantified using a ELISPOT kit. Percentages of CD4＋/CD8＋ effector cells or memory cells in the spleen cells of mice were analyzed with flow cytometer. Results The mice of all groups induced specific IgG antibodies compared with PBS group after the third immunization （ P 〈0.05） ,except the level of IgG in IL-23＋G1F/M2 group is the lowest. The specific lysis activity induced by IL-23＋CpGzz,6 ＋G1F/M2 was significantly higher than other groups. 1L-23＋ CPG2216＋G1F/M2group, induced significantly more IFN 7 secreting cells than other groups （ P〈0.05）. In addition, the level of IFN-γ secreting cells was remarkably higher than that of IL-4-secreting cells in each group. Both IL-23 and IL-23＋CpG2216, have significant advantages to enhanced immune memory response. Conclusions IL 23 ＋ Cp2216 , as an adjuvant, can enhanced significantly the cellular immune responses of RSV recombinant vaccine GIF/M2.

关 键 词：白介素23 CpG2216佐剂 呼吸道合胞病毒 重组疫苗 

分 类 号：R3[医药卫生—基础医学]