摘要
目的研究细胞因子佐剂白介素27（IL-27）及佐剂白介素23（IL-23）与佐剂CpG2216对呼吸道合胞病毒（RSV）重组疫苗（G1F/M2）免疫原性的影响。方法质粒IL-27、IL-23与佐剂CpG2216单独或联合与疫苗G1F/M2共免疫Balb/c小鼠3次,每次免疫后10 d取血,分离血清,用间接ELISA法测定3次免疫后的血清IgG抗体效价,第3次免疫后2周处死小鼠,取脾细胞,用LDH法检测CTL活性,ELISPOT法检测分泌IL-4和IFN-γ的淋巴细胞水平,流式细胞仪检测CD4＋及CD8＋细胞的效应记忆和中央记忆T细胞。结果 ELISA结果表明：除PBS组外,各组均诱导了高水平的IgG抗体,与PBS＋G1F/M2组相比,只有IL-23＋G1F/M2组的IgG水平是显著降低的,而其余各组相比无显著性差异;CTL杀伤实验显示：IL-23＋CpG2216＋G1F/M2组和IL-23＋IL-27＋G1F/M2组的CTL杀伤活性显著高于其他各组（P〈0.05）,IL-23＋CpG2216＋G1F/M2组的CTL最为显著;ELISPOT结果显示：IL-23＋IL-27＋G1F/M2组、IL-23＋CpG2216＋G1F/M2组、IL-27＋CpG2216＋G1F/M2组中分泌IFN-γ的淋巴细胞数量显著高于其他组分泌IFN-γ的淋巴细胞数量,且各组分泌IFN-γ的淋巴细胞数量显著高于同组分泌IL-4的淋巴细胞数量（P〈0.05）;流式细胞仪检测结果说明IL-23＋IL-27对增强G1F/M2诱导记忆应答方面具有显著优势。结论佐剂IL-23、IL-27与CpG2216联合使用均能增强RSV重组疫苗的免疫应答。
To evaluate the effects of IL-23, IL-27 and CPG2216 as adjuvants on the cellular immune response induced by recombinant protein vaccine of respiratory syncytial virus （RSV）, Balb/c mice were immunized with pcDNA3.1/IL-23, pcDNA3.1/IL-27 alone or in combination with RSV vaccines G1F/M2 three times at an interval of 10 days. Mice were phlebotomized at day 10 after every immunization and the serum was separated for measuring specific antibody IgG titers by indirect ELISA. Two weeks after the third immunization, the mice were sacrificed and splenocytes were prepared for assessing specific lysis activity using a LDH Cytotoxicity Assay Kit. Furthermore, IFN- γ and IL--4 secreting cells were quantified using a ELISPOT kit, while the percentage of CD4＋ ／CD8＋ effector cells or memory cells in the spleen cells was analyzed with Flow Cytometer. We found that the mice in all groups, except PBS group, induced specific IgG antibodies after the third immunization; as compared with PBS＋G1F/M2 group, IL-23＋ G1F/M2 group demonstrated significant lower IgG level, but other group showed no significant difference. The CTL activity of IL-23 ＋CpG=2216＋G1F/M2 group and IL-23 ＋IL-27 ＋G1F/M2 groups, especially IL-23 ＋CPG=I6＋G1F/M2 group （P〈0.05）, was significant higher than that of the other groups. IL-23＋IL-27＋G1F/M2, IL-23＋CpGz2216＋G1F/M2 and IL-27＋CpG=16＋G1F/M2 groups, especially IL-23＋CPGzz16＋G1F/M2 group, induced significantly more INF-7 secreting cells than other groups did （P 〈 0.05）. These results indicated that IL-23 ＋IL-27 have significant advantages to enhance immune memory response. The adjuvant IL-23/IL-27 with CPG2216 could enhance significantly the cellular immune responses of RSV recombinant vaccine G1F/M2.
出处
《免疫学杂志》
CAS
CSCD
北大核心
2013年第7期566-571,共6页
Immunological Journal
基金
国家自然科学基金（30972801）
