Liu Ye, Li Fusheng, Qi Zhi, Hao Yanling, Hong Kunxue, Liu Yong, Cong Yulong, Shao Yiming
Department of Clinical Laboratory, Chinese P, L, A, General Hospital, No, 28 Fuxing Road, Beijing 100853, China.
Virol J. 2013 Sep 30;10:297. doi: 10.1186/1743-422X-10-297.
HIV trans-activator protein (Tat) is the crucial factor to control HIV transcription, and is usually considered as an important immunogen for the design of HIV vaccine. Recent studies reported some special bio-activities of Tat protein on immunoregulation. However, to date, few studies have focused on exploring the effects of Tat expression plasmid (pTat) on regulating the immune responses induced by HIV DNA vaccines. In this study, our main objective is to investigate the immunoregulation mediated by pTat in mice.
Four gene-coding plasmids (pTat, pGag, pEnv and pPol) were constructed, and the gene expression was detected by western blot method. The effects of pTat on regulating the immune responses to antigens Gag, Env, Pol were assessed by enzyme-linked immunospot and enzyme-linked immunosorbent assay. The data was analysed by one-way analysis of variance.
After two immunizations, mice vaccinated with antigen expressing plasmid (pGag, pEnv or pPol) plus pTat exhibited significantly stronger IFN-gamma response than that vaccinated with the corresponding antigen alone. Moreover, mice receiving two injections of antigen plus pTat exhibited the same strong IFN-gamma response as those receiving three injections of antigen alone did. Furthermore, addition of pTat not only induced a more balanced Th1 and Th2 response, but also broadened IgG subclass responses to antigens Gag and Pol.
pTat exhibited the appreciable effects on modulating immune responses to HIV antigens Gag, Env and Pol, providing us interesting clues on how to optimize HIV DNA vaccine.
HIV反式激活蛋白(Tat)是控制HIV转录的关键因子,通常被视为HIV疫苗设计的重要免疫原。最近的研究报道了Tat蛋白在免疫调节方面的一些特殊生物活性。然而,迄今为止,很少有研究关注探索Tat表达质粒(pTat)对调节HIV DNA疫苗诱导的免疫反应的影响。在本研究中,我们的主要目的是研究pTat在小鼠中介导的免疫调节作用。
构建了四种基因编码质粒(pTat、pGag、pEnv和pPol),并通过蛋白质免疫印迹法检测基因表达。通过酶联免疫斑点法和酶联免疫吸附测定法评估pTat对调节针对抗原Gag、Env、Pol的免疫反应的影响。数据采用单因素方差分析。
两次免疫后,接种抗原表达质粒(pGag、pEnv或pPol)加pTat的小鼠比单独接种相应抗原的小鼠表现出明显更强的IFN-γ反应。此外,接受两次抗原加pTat注射的小鼠与单独接受三次抗原注射的小鼠表现出相同强烈的IFN-γ反应。此外,添加pTat不仅诱导了更平衡的Th1和Th2反应,还拓宽了针对抗原Gag和Pol的IgG亚类反应。
pTat对调节针对HIV抗原Gag、Env和Pol的免疫反应具有显著作用,为我们优化HIV DNA疫苗提供了有趣的线索。