乙肝疫苗新型佐剂CpG-BW006对小鼠脾NK细胞表面分子CD69的表达及γ干扰素分泌的影响

Influences of CpG Adjuvant BW006 of Hepatitis B Vaccine on Mu rine Spleen NK Cells Surface CD69 Expression and IFN-γ Secretion

目的:研究重组乙型肝炎表面抗原(HBsAg)佐剂BW006对小鼠脾自然杀伤(NK)细胞表面分子CD69的表达和γ干扰素(IFN-γ)分泌水平的影响。方法:BW006、HBsAg单用或联用体外刺激小鼠脾NK细胞,流式细胞仪检测NK细胞膜表面分子CD69的表达水平,ELISA检测IFN-γ的分泌水平。结果:5μg BW006体外刺激小鼠脾NK细胞24 h后,NK细胞表面分子CD69的表达达峰值(阳性率45.18%),显著高于40μg HBsAg组(21.44%)(P<0.05),与5μg BW006和40μg HBsAg联用组(58.49%)相比无显著差异;24 h时,5μg BW006组的IFN-γ分泌水平达56.95ng/mL,显著高于40μg HBsAg组(8.74 ng/mL)(P<0.05),与联用组(57.70 ng/mL)相比无显著差异。结论:BW006具有上调NK细胞表面分子CD69表达和诱导IFN-γ分泌的早期活化NK细胞的功能,作为疫苗的新型佐剂前景较好。

Objective： To study the influences of CpG adjuvant BW006 for hepatitis B vaccine on the pheno- type and functions of murine spleen natural killer（NK） cells. Methods： After murine spleen NK cells were stimu- lated with BW006 and/or HBsAg in vitro, the expression of CD69 on NK cells surface was analyzed by flow cy- tometer and the secretion level of IFN-~/ was detected by ELISA. Results： After 24 hours of stimulation, the ex- pression of CD69 reached peak and the expression rate was 48.18% which was significantly higher than 40 p,g HBsAg group（21.44%, P〈0.05）. However, the expression of CD69 on NK cells surface have no significant differ- ence in 5 p,g BW006 group and 5 ug BW006 combined with 40 p,g HBsAg group（58.49%）. The IFN-＇y secre- tion levels of murine spleen NK cells was up to 56.95 ng/mL after activating with 5 p^g BW006 which was signifi- cantly higher than 40 Ixg HBsAg group （8.47 ng/mL, P〈0.05）. However, the IFN-y secretion levels of routine spleen NK cells have no significant difference in 5 p,g BW006 group and 5 μg BWO06 combined with 40 ＆g HB- sag group（57.70%）. Conclusion： BW006 functions to activate NK cells on early stage, regulate expression of CD69 and induce IFN-y secretion, indicating BW006 to be a novel promising vaccine adjuvant.