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目的研究细胞因子佐剂IL-27单独或与Cp G2216佐剂联合使用对呼吸道合胞病毒(RSV)重组蛋白疫苗G1F/M2免疫病理的调节作用。方法将编码IL-27的质粒(以下简称IL-27)单独或与Cp G2216佐剂联合使用与G1F/M2共免疫Balb/c小鼠3次,第3次免疫后第10天用RSV攻击免疫后的Balb/c小鼠,攻毒后5 d处死,留取肺组织,用实时定量PCR检测RSV-N基因的表达量,Th1和Th2型细胞因子IFN-γ和IL-5,中性粒细胞和嗜酸性粒细胞趋化因子Gro-α和eotaxin,Th1和Th2特异性转录因子T-bet和GATA3及黏液因子gob-5的m RNA的表达量;另一部分肺组织做病理切片,用HE染色观察肺组织情况。结果 PBS组明显被RSV感染,而各疫苗组显著被保护。IL-27+G1F/M2组和IL-27+Cp G2216+G1F/M2组的IFN-γ表达量无差异(P>0.05),且均显著高于无佐剂的G1F/M2组(P<0.05);IL-27+Cp G2216+G1F/M2组的T-bet表达量显著高于其它组(P<0.05);IL-27+Cp G2216+G1F/M2和IL-27+G1F/M2组的GATA3、IL-5表达量均显著低于G1F/M2组(P<0.05),且IL-27+G1F/M2组的IL-5表达量在疫苗组中最低(P<0.05);IL-27+G1F/M2组的Gro-α、eotaxin、gob-5表达量显著低于其它疫苗组(P<0.05)。病理切片结果显示:与正常小鼠相比,各组经RSV攻击后肺组织均有不同程度的炎症病理损伤,而IL-27+G1F/M2组的肺部的炎症病理损伤与其它各组相比明显减轻。结论 IL-27+Cp G对疫苗增强性肺部免疫病理有一定的下调作用,而IL-27能显著抑制疫苗增强性肺部免疫病理。
Objective To investigate the regulatory effect of cytokine adjuvant IL-27 alone or in combination with Cp G2216 adjuvant on the immunopathology of respiratory syncytial virus (RSV) recombinant protein vaccine G1F / M2. Methods Balb / c mice co-immunized with G1F / M2 alone or in combination with Cp G2216 adjuvant were used to immunize Balb / c mice with the IL-27-encoding plasmid (hereinafter referred to as IL-27) for 10 days after the third immunization The Balb / c mice were sacrificed 5 d after challenge and the lung tissue was collected. The expression of RSV-N gene, Th1 and Th2 cytokines IFN-γ and IL-5, Cells and eotaxin Gro-α and eotaxin, Th1 and Th2-specific transcription factor T-bet and GATA3 and mucus gob-5 m RNA expression; the other part of the lung tissue biopsy with HE Staining observed lung tissue. Results The PBS group was significantly infected with RSV, while each vaccine group was significantly protected. There was no difference in IFN-γexpression between IL-27 + G1F / M2 group and IL-27 + Cp G2216 + G1F / M2 group (P> 0.05) 0.05). The expression of T-bet in IL-27 + Cp G2216 + G1F / M2 group was significantly higher than that in other groups (P <0.05); IL-27 + Cp G2216 + G1F / (P <0.05), and the expression of IL-5 in IL-27 + G1F / M2 group was the lowest (P <0.05) The expression of Gro-α, eotaxin and gob-5 in 27 + G1F / M2 group was significantly lower than that in other vaccine groups (P <0.05). The results of pathological examination showed that compared with normal mice, the lung tissues of all groups had different degrees of inflammatory pathological changes after RSV challenge, while the inflammatory pathological changes of lung in IL-27 + G1F / M2 group were significantly different from those in other groups Than the obvious reduction. Conclusion IL-27 + Cp G can down-regulate the immune pathology of the vaccine-enhanced lungs, while IL-27 can significantly inhibit the immunopathology of the vaccine-enhanced lungs.