Zhang Lishu, Ningyi Jin, Yingjin Song, Yansong Sun, Hong Wang, Dawei Zhan, Ma Hewen, Shang Yupu, Jin Hongtao, Hong Baoqing, Li Chang
1The 11th Institute, Academy of Military Medical Sciences, Changchun, 130062 China.
2College of Agricultural and Biological Engineer, Tianjin University, Tianjin, 30072 China.
Chin Sci Bull. 2006;51(13):1571-1577. doi: 10.1007/s11434-006-1571-9.
An effective HIV-1 vaccine will be the ultimate solution for the prevention of HIV/AIDS, though HAART plays important roles in treating the disease. In this study, a large-scale recombinant DNA plasmid containing a designed HIV-1 multi-epitope-p24 chimeric gene was prepared and purified. Rhesus monkeys were then inoculated muscularly with the plasmid for four times in week 0, 4, 8 and 18. Whole blood was collected two weeks after the third and fourth inoculation, followed by serum and peripheral blood mononuclear cell (PBMC) separation. The CTL activity and proliferation of PBMCs stimulated by macaque MHC-I-restricted HIV-1 CTL epitope peptide were analyzed by MTT and LDH release assay, respectively. Th1 cytokines in supernatant of cultured PBMC stimulated by HIV-1 CTL epitope peptide and anti-HIV-1 antibody in serum were assayed by ELISA. The results showed that increased CTL target-killing activity, higher secretion of Th1 cytokines (IFN-γ and IL-2) and promoted proliferative reaction of monkey PBMCs stimulated by HIV-1 CTL epitope peptide were detected in the immunization group inoculated by the recombinant DNA vaccine for three times, which were further enhanced by the fourth inoculation. At the same time, HIV-1 specific antibody in serum of immunized monkeys was higher than that in controls. We concluded that the designed HIV-1 DNA vaccine may induce HIV-1 specific cellular and humoral immunity on monkeys.
尽管高效抗逆转录病毒疗法（HAART）在治疗艾滋病方面发挥着重要作用，但有效的HIV-1疫苗将是预防HIV/AIDS的最终解决方案。在本研究中，制备并纯化了一种含有设计的HIV-1多表位-p24嵌合基因的大规模重组DNA质粒。然后在第0、4、8和18周对恒河猴进行四次肌肉注射该质粒。在第三次和第四次接种后两周采集全血，随后分离血清和外周血单个核细胞（PBMC）。分别通过MTT法和LDH释放试验分析猕猴MHC-I限制性HIV-1 CTL表位肽刺激的PBMC的CTL活性和增殖情况。通过ELISA检测HIV-1 CTL表位肽刺激培养的PBMC上清液中的Th1细胞因子和血清中的抗HIV-1抗体。结果显示，在接种重组DNA疫苗三次的免疫组中，检测到HIV-1 CTL表位肽刺激的猴PBMC的CTL靶细胞杀伤活性增强、Th1细胞因子（IFN-γ和IL-2）分泌增加以及增殖反应增强，第四次接种后进一步增强。同时，免疫猴血清中的HIV-1特异性抗体高于对照组。我们得出结论，所设计的HIV-1 DNA疫苗可能在猴体内诱导HIV-1特异性细胞免疫和体液免疫。
