C3d3通过促进脾脏B细胞高表达CXCR4增强hCGβ DNA免疫体液免疫效应  

GENE CONJUGATION OF MOLECULAR ADJUVANT C3d3 TO hCGβ ENHANCED THE ANTI-hCGβ HUMORAL IMMUNE RESPONSE VIA UPREGULATION OF CXCR4 EXPRESSION ON SPLENIC B CELLS IN DNA VACCINATION

作　　者：贺晓菊[1] 李大金[1] 姚晓英[1] 袁敏敏[1] 

机构地区：[1]复旦大学上海医学院附属妇产科医院暨妇产科研究所生殖免疫研究室,上海200011

出　　处：《分子细胞生物学报》2007年第4期185-192,共8页Journal of Molecular Cell Biology

基　　金：本文受国家自然科学基金(30271235);上海市自然科学基金资助(00zB14058)

摘　　要：本文旨在探讨分子佐剂C3d3与hCGβ融合在基因免疫中增强抗hCGβ体液免疫效应的机制。分别用质粒pCMV4-hCGB-C3d3、pCMV4-hCGβ和pCMV4免疫BALB/c小鼠,间接ELISA法检测免疫小鼠外周血IgG/IgA类抗hCGβ抗体水平;ELISPOT分析免疫鼠脾脏组织IgG/IgA类抗体分泌细胞水平(ASC);RT-PCR分析免疫鼠脾脏B细胞趋化因子受体表达,RT-PCR和FCM分析CXCR4表达水平;RT-PCR和ELISA检测脾脏组织CXCL12表达水平。结果显示,pCMV4- hCGβ-C3d3免疫组外周血IgG类抗hCGβ抗体水平明显高于pCMV4-hCGβ免疫组;而IgA类抗hCGβ抗体水平在两组间无明显差异。pCMV4-hCGβ-C3d3免疫组脾脏组织IgG类ASCs水平明显高于pCMV4-hCGβ组;两组间IgA类ASCs水平无明显差异。经pCMV4-hCGB、pCMV4-hCGβ- C3d3免疫鼠脾脏B细胞CXCR4表达明显高于对照组;且pCMV4-hCGβ-C3d3组明显高于pCMV4-hCGβ免疫组。CXCR4^+细胞与ASCs呈正相关,r=0.966,(P<0.05)。pCMV4-hCGβ-C3d3和pCMV4-hCGβ组脾脏组织CXC L12表达均显著高于对照组。结果表明,分子佐剂C3d3与hCGβ基因融合,在基因免疫小鼠后能够显著升调节脾脏ASCs CXCR4表达,从而可能增强抗hcGβ基因疫苗的体液免疫效应。To investigate mechanism of the anti-hCGβ humoral immune responsive enhancement by gene conjugation of the molecular adjuvant C3d3 to hCGβ in DNA immunization, BALB/c mice were inoculated intramuscularly with pCMV4-hCGβ-C3d3, pCMV4-hCGβ and pCMV4, respectively. The titers of anti-hCGβ IgG/IgA antibody in serum were determined by indirect ELISA. The IgG/IgA ASCs levels were evaluated by ELISPOT. The expressions of chemokine receptors on B cells were analyzed by RT-PCR, and CXCR4 expression was analyzed respectively by RT-PCR and FCM. The expression of CXCL12 in spleen was investigated respectively by RT-PCR and ELISA. We found that anti-hCGβ IgG antibody titer in serum after pCMV4-hCGβ-C3d3 immunization was significantly higher than that of pCMV4-hCGβ immunization. But the anti-hCGβ IgA antibody titers appeared no difference between the two immunized groups. The level of IgG ASCs in spleen of pCMV4-hCGβ-C3d3 immunization was significantly higher than that of pCMV4-hCGβ immunization,but the levels of IgA ASCs appeared no difference between the two groups. The expression of CXCR4 on B cell increased after pCMV4-hCGβ-C3d3 immunization, and significantly higher than that of pCMV4-hCGβ immunization. The rate of CXCR4^＋ cell was correlated to the numbers of the ASC （r=-0.966, P〈0.05）. CXCL12 expression increased after pCMV4-hCGβ and pCMV4-hCGβ-C3d3 immunization, and appeared no difference between the two groups. These resuits above indicate that gene fusion of the molecular adjuvant C3d3 to hCGβ can significantly enhance the antigen-specific antibody response, and the level of IgG ASCs in spleen via upregulation of CXCR4 expression on splenic B cells in DNA vaccination.

关 键 词：分子佐剂C3d3 hCGβ基因免疫 CXCRR/CXCL12 

分 类 号：R392[医药卫生—免疫学]