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目的研究共刺激分子4-1BBL基因免疫对HBsAg核酸疫苗诱导小鼠特异性体液和细胞免疫应答的影响。方法将HBV表面抗原核酸疫苗pcDS2单独或联合共刺激分子4-1BBL质粒肌肉注射免疫C57BL/6小鼠;ELISA法检测小鼠血清抗-HBsIgG及亚型IgG1和IgG2a;迟发型超敏反应(DTH)反应检测体内细胞反应;流式细胞仪检测CD4+T淋巴细胞分泌IL-4和IFN-γ及CD8+T淋巴细胞分泌IFN-γ水平;流式细胞仪检测小鼠脾细胞HBsAg特异性体外细胞毒性T淋巴细胞杀伤作用(CTL)。结果与单纯免疫核酸疫苗pcDS2组比较,pcDS2和4-1BBL联合免疫组小鼠的抗-HBs水平显著提高,抗-HBsIgG亚类以IgG2a占优;免疫小鼠经HBsAg脚掌皮下刺激后,联合免疫组小鼠脚掌的厚度显著高于pcDS2组;联合免疫组CD4+T淋巴细胞的IL-4和IFN-γ表达水平及CD8+T淋巴细胞的IFN-γ表达水平显著升高;DNA疫苗免疫的各组小鼠,HBsAg特异性体外CTL杀伤作用高于对照组,其中联合免疫组小鼠的体外CTL杀伤作用最强。结论共刺激分子4-1BBL不仅能增强HBV DNA疫苗诱导特异性体液免疫应答,还能增强特异性型细胞免疫反应,尤其增强体内CTL的杀伤活性。
Objective To investigate the effect of 4-1BBL gene immunization on specific humoral and cellular immune responses induced by HBsAg DNA vaccine in mice. Methods C57BL / 6 mice were immunized intramuscularly with HBV surface antigen DNA vaccine pcDS2 alone or in combination with costimulatory molecule 4-1BBL. Serum anti-HBs IgG and IgG1 and IgG2a were detected by ELISA. Delayed-type hypersensitivity (DTH) ) Were detected by flow cytometry. The levels of IL-4 and IFN-γ secreted by CD4 + T lymphocytes and the secretion of IFN-γ by CD8 + T lymphocytes were detected by flow cytometry. The levels of HBsAg secreted by mouse spleen cells were detected by flow cytometry Cytotoxic T lymphocyte cytotoxicity (CTL). Results Compared with the immunized DNA vaccine pcDS2 group, the level of anti-HBs in pcDS2 and 4-1BBL combined immunized mice was significantly increased, and the anti-HBs IgG subclass was dominated by IgG2a. The immunized mice were immunized subcutaneously with HBsAg soles The thickness of soles of mice in group A was significantly higher than that in pcDS2 group. The expression of IL-4 and IFN-γ in CD4 + T lymphocytes and the expression of IFN-γ in CD8 + T lymphocytes were significantly increased in combination group. In each group, HBsAg-specific cytotoxicity against CTL in vitro was higher than that in control group, especially in combination CTL mice. Conclusion Co-stimulatory molecule 4-1BBL not only enhances the specific humoral immune response induced by HBV DNA vaccine, but also enhances the specific type of cellular immune response, especially enhancing CTL killing activity in vivo.