Jiang Wenzheng, Ren Lisong, Jin Ningyi
Genetic Engineering Key Laboratory of PLA, The Eleventh Institute of Academy of Military Medical Sciences of PLA, Changchun 130062, People's Republic of China.
J Virol Methods. 2007 Dec;146(1-2):266-73. doi: 10.1016/j.jviromet.2007.07.016. Epub 2007 Sep 14.
Numerous strategies have been employed in an attempt to improve the immunogenicity and efficacy of nucleic acid vaccines. In the present study, the immunogenicity in the induction of humoral and cellular immune responses to HIV-1 DNA vaccine expressing a chimeric gene of gag and gp120 and the adjuvant effect of IFN-alpha on HIV-1 DNA vaccine were studied in a murine model. The DNA vaccine plasmid pVAX1-gag-gp120 and eukaryotic expression plasmid pVAX1-IFN were constructed by inserting the chimeric gene of gag and gp120 of HIV-1 and IFN-alpha into the downstream of CMV promoter of eukaryotic expression vector pVAX1, respectively. In vitro expression detected by RT-PCR and Western blotting showed that the genes of interest could be expressed in transfected HeLa cells. After BALB/c mice were immunized by three intramuscular inoculations of the HIV-1 DNA vaccine plasmids alone or in combination with IFN-alpha expression plasmids, the different levels of anti-HIV-1 humoral and cellular responses were measured comparable to the control groups immunized with pVAX1-IFN, parent plasmid pVAX1 or PBS. The percentage of CD3+CD4+ and CD3+CD8+ subgroups of spleen T lymphocytes and the specific cytotoxicity activities of splenic CTLs in the coinoculation group were significantly higher than those in the separate inoculation group, and an enhancement of antibody response was also observed in the coinoculation group compared with the separate inoculation group. Take together, coadministration of HIV-1 DNA vaccine plasmids and IFN-alpha expression plasmids can elicit stronger humoral and cellular immune responses in mice than HIV-1 DNA vaccine plasmids alone, and IFN-alpha can be an effective immunological adjuvant in DNA vaccination against HIV-1.
为提高核酸疫苗的免疫原性和效力,人们采用了多种策略。在本研究中,在小鼠模型中研究了表达gag和gp120嵌合基因的HIV-1 DNA疫苗诱导体液和细胞免疫反应的免疫原性以及IFN-α对HIV-1 DNA疫苗的佐剂作用。分别将HIV-1的gag和gp120嵌合基因以及IFN-α插入真核表达载体pVAX1的CMV启动子下游,构建DNA疫苗质粒pVAX1-gag-gp120和真核表达质粒pVAX1-IFN。通过RT-PCR和Western印迹检测的体外表达表明,目的基因可在转染的HeLa细胞中表达。用HIV-1 DNA疫苗质粒单独或与IFN-α表达质粒联合经三次肌肉注射免疫BALB/c小鼠后,检测到不同水平的抗HIV-1体液和细胞反应,与用pVAX1-IFN、亲本质粒pVAX1或PBS免疫的对照组相当。联合接种组脾脏T淋巴细胞的CD3 + CD4 +和CD3 + CD8 +亚群百分比以及脾脏CTL的特异性细胞毒性活性均显著高于单独接种组,且联合接种组与单独接种组相比抗体反应也有所增强。综上所述,HIV-1 DNA疫苗质粒与IFN-α表达质粒联合给药在小鼠中能引发比单独使用HIV-1 DNA疫苗质粒更强的体液和细胞免疫反应,且IFN-α可作为抗HIV-1 DNA疫苗接种的有效免疫佐剂。