肥大细胞活化剂compound 48/80联合乙肝疫苗对小鼠免疫功能的影响

The effect of mast cell activator compound 48/80 on hepatitis B vaccine-induced immune response

目的初步探讨肥大细胞活化剂compound 48/80(C48/80)联合乙肝疫苗对实验小鼠抗体产生、淋巴细胞增殖的影响,寻找能够增强机体免疫应答能力的佐剂。方法 Balb/c小鼠经肩背部皮下注射C48/80,甲苯胺蓝法检测肥大细胞活化状态,流式细胞术检测肩上和腋窝局部引流淋巴结(draining lymph nodes,DLN)细胞悬液中树突状细胞(dendritic cell,DC)的数量;采用3种剂量C48/80联合乙肝疫苗免疫小鼠,ELISA法检测不同时间点血清乙肝表面抗体(抗-HBs)的含量,CCK-8试剂检测DLN淋巴细胞增殖活性。结果皮下注射C48/80可诱导小鼠局部皮肤肥大细胞活化,24 h脱颗粒肥大细胞数占局部肥大细胞总数比例为82%±5.2%;C48/80处理后DLN细胞总数和DC数量较对照组明显增多。C48/80联合乙肝疫苗免疫小鼠后3周和4周时血清中抗-HBs含量较单纯乙肝疫苗免疫组明显升高;C48/80处理组小鼠特异性淋巴细胞增殖能力较单纯乙肝疫苗免疫组明显增强。结论肥大细胞活化剂C48/80能够在较早时间点增强小鼠对乙肝疫苗的免疫应答,其机制可能与C48/80诱导DLN DC细胞数量增多有关。

The present study was designed to investigate the adjuvant efficacy of compound 48/80（C48/80） in hepatitis B vaccine immunized mice. Firstly, C48/80 was injected into Balb/c mouse scapular site to induce local mast cell degranulation; and dendritic cells in the draining brachial and axillary lymph nodes were detected by FACS. Then Balb/c mice were vaccinated with hepatitis B vaccine and C48/80; and serum anti-HBs and specific lymphocyte proliferation ability were detected by ELISA and CCK-8 kit respectively. Data showed that C48/80 could induce obvious mast cell degranulation, with an 82%±5.2% degranulated mast cell percentage at 24 h. Total cells and dendritic cells in draining lymph nodes were both increased by C48/80 treatment. In contrast to simple hepatitis B vaccine immunization group, anti-HBs concentrations were significantly higher in mice vaccinated with hepatitis B vaccine plus C48/80 at week 3 and week 4, specific lymphocyte proliferation ability was also enhanced in C48/80 immunized mice. In conclusion, C48/80 could boost the immune response of hepatitis B vaccine in relatively earlier time, which may be related with C48/80 induced dendritic cell recruitment in draining lymph nodes.