论文部分内容阅读
目的研究细胞因子佐剂IL-27单独或与CpG2216佐剂联合使用对呼吸道合胞病毒(RSV)重组蛋白疫苗免疫效果的影响。方法将编码IL-27的质粒(以下简称IL-27)单独或与CpG2216佐剂作为联合佐剂与G1F/M2共免疫Balb/c小鼠3次,每次免疫10 d后取血,分离血清,用间接ELISA法测定3次免疫后的血清IgG抗体效价以及末次IgG1、IgG2a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~+、CD8~+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抗体无显著性差异(P<0.05),IgG2a、IgG1分别为Th1和Th2型抗体,IL-27+G1F/M2组、IL-27+CpG2216+G1F/M2组IgG2a和IgG1的比值明显高于其他组,说明IL-27作为佐剂对G1F/M2所诱导的体液免疫应答无增强作用但可以调节免疫应答的类型,使免疫应答更平衡;CTL杀伤实验显示:IL-27~+CpG2216+G1F/M2组的CTL杀伤活性显著高于其他各组(P<0.05);ELISPOT结果显示: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),即均诱导了Th1型优势应答;流式细胞仪检测结果说明IL-27+G1F/M2组和IL-27~+CpG2216+G1F/M2组均能诱导产生大量的CD44~+CD62L~+双阳性细胞。结论细胞因子佐剂IL-27和CpG2216佐剂联合使用可以增强吸道合胞病毒重组疫苗诱导的细胞免疫应答而且IL-27对Th1型优势应答具有调节作用。
Objective To investigate the effect of cytokine adjuvant IL-27 alone or in combination with CpG2216 adjuvant on the immunogenicity of recombinant respiratory syncytial virus (RSV) vaccine. Methods Balb / c mice were co-immunized with IL-27 plasmid (IL-27) alone or with CpG2216 adjuvant for three times with G1F / M2. After each immunization for 10 days, blood was collected and the serum was separated The serum IgG antibody titers of the three immunizations and the last IgG1 and IgG2a antibody titers were measured by indirect ELISA. The mice were sacrificed two weeks after the third immunization. The splenocytes were harvested and the activity of CTL was detected by LDH. The levels of IL-4 and IFN-γ secreting lymphocytes were detected by ELISPOT. The levels of CD4 ~ + and CD8 ~ + were detected by flow cytometry The effect of memory and central memory T cells. Results The results of ELISA showed that there was no significant difference (P <0.05) between high and low level of IgG antibodies in all groups except PBS group. The levels of IgG2a and IgG1 were Th1 and Th2 antibodies, IL-27 + G1F / M2 group, The ratio of IgG2a to IgG1 in IL-27 + CpG2216 + G1F / M2 group was significantly higher than that in other groups, indicating that IL-27 did not enhance the humoral immune response induced by G1F / M2 but could regulate the type of immune response The CTL killing test showed that the cytotoxic activity of CTL in IL-27 ~ + CpG2216 + G1F / M2 group was significantly higher than that in other groups (P <0.05). ELISPOT results showed that IL-27 ~ + CpG2216 + G1F / The number of lymphocytes secreting IFN-γ in M2 group was significantly higher than that in other groups, and the number of lymphocytes secreting IFN-γ in each group was significantly higher than that of IL-4 secreting lymphocytes in the same group P <0.05), that is, Th1-type predominant response was induced. The results of flow cytometry showed that both IL-27 + G1F / M2 group and IL-27 + CpG2216 + G1F / M2 group could induce a large number of CD44 + CD62L ~ + double positive cells. Conclusions The combination of cytokine adjuvant IL-27 and CpG2216 adjuvant can enhance the cellular immune response induced by recombinant vaccine of respiratory syncytial virus and IL-27 has a regulatory effect on Th1-type predominant response.