HPV6L1/CtMOMP多表位融合基因在COS-7细胞的表达及其在小鼠的免疫效果观察

研究人乳头瘤病毒(human papillomavirus,HPV)6b结构蛋白L1(HPV6b L1)基因佐剂增强沙眼衣原体(Chlamydia trachomatis,Ct)主要外膜蛋白(MOMP)多表位(Ct MOMP168)DNA疫苗的免疫效果的可能性.构建pcDNA3.1(+)/HPV6b L1/Ct MOMP168融合重组质粒,转染COS-7细胞,用RT-PCR、激光共聚焦显微技术及Western印迹技术检测其表达.分别用pcDNA3.1(+)/HPV6b L1/Ct MOMP168、pcDNA3.1(+)/Ct MOMP168及pcDNA3.1(+)质粒肌肉免疫BALB/c小鼠,ELISA检测外周血中IgG及阴道分泌物中sIgA.结果表明,pcDNA3.1(+)/HPV6b L1/CtMOMP168可在COS-7细胞中表达;Ct MOMP168组和HPV6b L1/Ct MOMP168组均可刺激小鼠产生抗Ct MOMP特异性的抗体,抗体滴度随免疫次数增加而升高,且HPV6b L1/Ct MOMP168组小鼠产生的抗体滴度明显高于Ct MOMP168组(P0.05).结果提示,分子佐剂HPV6b L1与CtMOMP多表位基因融合能够显著增强Ct MOMP多表位DNA疫苗的体液免疫应答.

Expression of Fusion Gene Coding for Protective Antigen Ct MOMP Multi-epitope and HPV 6b L1 in COS-7 Cells and Its Immunogenicity in Mice

To investigate the possible enhancement of the immunological effects of a DNA vaccine to the Chlamydia trachomatis (Ct) major outer membrane protein (MOMP) multi-epitope gene (Ct MOMP 168) using the human papillomavirus (HPV) type 6b capsid protein L1 (HPV 6b L1) gene, COS-7 cells were transfected with the plasmid pcDNA3.1(+)/HPV 6b L1/Ct MOMP 168, which co-expressed HPV 6b L1 and Ct MOMP 168, or pcDNA3.1(+). Western immunoblotting, confocal microscopy and RT-PCR were used to detect the expression of HPV 6b L1-Ct MOMP 168. BALB/c mice were inoculated intramuscularly with pcDNA3.1(+)/HPV 6b L1/Ct MOMP 168, pcDNA3.1(+)/Ct MOMP 168 and pcDNA3.1(+). Sera and vaginal washes were collected for measuring IgG and secretory IgA (sIgA) antibodies, respectively. The results showed that HPV 6b L1-Ct MOMP 168 was expressed efficiently in COS-7 cells. Both pcDNA3.1(+)/HPV 6b L1/Ct MOMP 168 and pcDNA3.1(+)/ Ct MOMP 168 were able to induce production of anti-Ct IgG and sIgA antibodies in immunized mice, with the antibody titers in the pcDNA3.1(+)/HPV 6b L1/Ct MOMP 168 group higher (P<0.05). It is concluded that gene fusion of molecular adjuvant HPV 6b L1 to Ct MOMP 168 can significantly increase the antigen specific antibody response in Ct MOMP 168 DNA vaccine. It is concluded that gene fusion of molecular adjuvant HPV 6b L1 to Ct MOMP 168 can significantly increase the antigen specific antibody response in Ct MOMP 168 DNA vaccine.