Wang Fu-xiang, Sun Yong-tao, Sun Yong-nian, Xu Zhe, Wang Lin-xu, Bai Xue-fan, Huang Chang-xing
Center for Diagnosis & Treatment for Infectious Diseases of PLA, Tangdu Hospital, Fourth Military Medical Univercity, Xi'an 710038, China.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2004 May;20(3):282-5.
To construct the gp120 DNA vaccine of Chinese HIV-1 strain and evaluate the immune responses induced with it in BALB/c mice.
The recombinant expression vector pVAX1-GP120 was constructed by inserting HIV gp120 gene into the eukaryotic expression vector pVAX1 and confirmed with EcoR I/Pst I and DNA sequencing. BALB/c mice were immunized with pVAX1-GP120 and pVAX1 respectively. The levels of serum anti-HIV antibody and IFN-gamma of the immunized mice were detected by ELISA. The proliferation of splenocytes was determined by MTT colorimetry and the specific cytotoxic T lymphocytes (CTLs) response by LDH assay.
Restriction enzymes digestion analysis and DNA sequencing results revealed that the pVAX1-GP120 had been constructed successfully. The titer of anti-HIV antibody and the IFN-gamma level in mice immunized with the pVAX1-GP120 were higher than those in mice immunized with pVAX1 respectively (P<0.01). As compared with mice immunized with pVAX1 alone, the cytotoxic activity of specific CTLs and antigen-specific lymphoproliferative responses in mice immunized with pVAX1-GP120 were significantly enhanced (P<0.01).
Specific cellular and humoral immune responses in mice can be induced with gp120 gene vaccine of Chinese HIV-1 strain, which lays the foundation for further development of therapeutic HIV vaccine against HIV-1 infection.
构建中国HIV-1毒株的gp120 DNA疫苗,并评估其在BALB/c小鼠中诱导的免疫反应。
将HIV gp120基因插入真核表达载体pVAX1中构建重组表达载体pVAX1-GP120,经EcoR I/Pst I酶切和DNA测序鉴定。分别用pVAX1-GP120和pVAX1免疫BALB/c小鼠。用ELISA检测免疫小鼠血清抗HIV抗体水平和IFN-γ水平。用MTT比色法测定脾细胞增殖,用LDH法测定特异性细胞毒性T淋巴细胞(CTL)反应。
限制性内切酶消化分析和DNA测序结果显示pVAX1-GP120构建成功。用pVAX1-GP120免疫的小鼠抗HIV抗体效价和IFN-γ水平分别高于用pVAX1免疫的小鼠(P<0.01)。与单独用pVAX1免疫的小鼠相比,用pVAX1-GP120免疫的小鼠特异性CTL的细胞毒性活性和抗原特异性淋巴细胞增殖反应显著增强(P<0.01)。
中国HIV-1毒株的gp120基因疫苗可诱导小鼠产生特异性细胞免疫和体液免疫反应,为进一步研发抗HIV-1感染的治疗性HIV疫苗奠定了基础。