重组鸡白细胞介素-6/2融合蛋白对新城疫(LaSota株)活疫苗免疫增强作用研究

为探究重组鸡白细胞介素-6/2融合蛋白(rChIL-6-Linker-ChIL-2,重组融合蛋白)对新城疫病毒(NDV)活疫苗(LaSota株)的免疫增强作用,本研究将90只SPF鸡随机分为6组,分别为PBS对照组、NDV弱毒苗对照组、rChIL-6-Linker-ChIL-2免疫增强组、rChIL-6蛋白免疫增强组、rChIL-2蛋白免疫增强组及rChIL-6+rChIL-2混合蛋白免疫增强组,将鸡白细胞介素重组融合蛋白水剂与LaSota株联合接种于SPF鸡,分别采用MTS法、流式细胞术、ELISA和HA/HI法检测接种前后不同时间各组鸡外周血及脾淋巴细胞增殖活性、鸡外周血中CD3+CD4+/CD3+CD8+T淋巴细胞的百分含量、血清中Th1/Th2型细胞因子表达水平及免疫抗体水平等指标。结果显示,接种后7～21d时,与NDV弱毒苗对照组相比,同时接种重组融合蛋白组鸡淋巴细胞增殖活性、外周血CD3+CD4+/CD3+CD8+T淋巴细胞的百分含量比值及血清中重组鸡IL-4蛋白(rChIL-4)、ChIL-6、ChIL-2、重组鸡IFN-α蛋白(ChIFN-α)、ChIFN-γTh1/Th2型细胞因子表达水平明显提升;HI免疫抗体较NDV弱毒苗对照组提高了1.0～1.9个滴度,较单一rChIL-6、rChIL-2对照组分别提高了0.2～0.7、0.2～1.1个抗体滴度。综上所述,rChIL-6-Lin-ker-ChIL-2融合蛋白对NDV(LaSota株)活疫苗在鸡体内具有明显的免疫增强效果。

Immunopotentiation Effect of Recombinant Chicken Interleukin-6/2 Fusion Protein on Newcastle Disease Virus (LaSota) Live Vaccine

In order to study the immunopotentiation effect of recombinant chicken interleukin-6/2 fusion protein (rChIL-6-Linker-ChIL-2) on Newcastle disease vinus (NDV) live vaccine (LaSota vaccine),90 SPF chickens were chosen and randomly distributed into six groups which were the PBS control group,NDV live vaccine control group,rChIL-6-Linker-ChIL-2 immunopotentiation group,rChIL-6 protein immunopotentiation group,rChIL-2 protein immunopotentiation group and the rChIL-6+rChIL-2 fusion protein immunopotentiation group.The interleukin fusion protein fluid and NDV live vaccine (LaSota vaccine) were inoculated into SPF chickens.The indexes of proliferation activities of peripheral blood and splenic lymphocytes,percentages of the CD3⁺CD4⁺/CD3⁺CD8⁺ T-lymphocytes in peripheral blood,the contents of Th1/Th2-type cytokines and the specific immune antibody level were detected by assays of MTS,FCM,ELISA and HA/HI.The results showed that compared with the indexes of NDV vaccine inoculation control group,the lymphocytes proliferation activity,the percentage of CD3⁺CD4⁺/CD3⁺CD8⁺ T-lymphocytes,and the contents of Th1/Th2-type cytokines such as ChIL-4,ChIL-6,ChIL-2,ChIFN-α and ChIFN-γ of chickens in rChIL-6-Linker-ChIL-2 inoculation group were promoted obviously at 7 to 21 dpi,and the HI immune antibodies increased about 1.0 to 1.9 titer,which was 0.2 to 0.7 titer higher than that of rChIL-6 group and 0.2 to 1.1 titer higher than rChIL-2 that of group.In conclusion,the rChIL-6-Linker-ChIL-2 protein had an obvious immunopotentiation effect on NDV live vaccine (LaSota) in chickens.