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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

HE Xiao Ju~1 LI Da Jin~(1,2) YAO Xiao Ying~1 YUAN Min Min~1 (~1Laboratory for Reproductive Immunology,Hospital & Institute of Obstetrics and Gynecology,Shanghai Medical College,Fudan University,Shanghai 200011;~2Department of Obstetrics & Gynecology,The Affiliated Hospital of Hainan Medical College,Haikou 570102)  
To investigate mechanism of the anti-hCGβhumoral immune responsive enhance- ment by gene conjugation of the molecular adjuvant C3d3 to hCGβin DNA immunization,BALB/c mice were inoculated intramuscularly with pCMV4-hCGβ-C3d3,pCMV4-hCGβand pCMV4,re- spectively.The titers of anti-hCGβIgG/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-hCGβIgG antibody titer in serum after pCMV4-hCGβ-C3d3 immu- nization was significantly higher than that of pCMV4-hCGβimmunization.But the anti-hCGβIgA 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-hCGβimmunization,but the levels of IgA ASCs appeared no difference between the two groups.The ex- pression of CXCR4 on B cell increased after pCMV4-hCGβ-C3d3 immunization,and significantly higher than that of pCMV4-hCGβimmunization.The rate of CXCR4~+ cell was correlated to the numbers of the ASC (r=0.966,P0.05).CXCL12 expression increased after pCMV4-hCGβand pCMV4-hCGβ-C3d3 immunization,and appeared no difference between the two groups.These re- suits above indicate that gene fusion of the molecular adjuvant C3d3 to hCGβcan significantly en- hance 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.
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