Department of Hepatitis and AIDS, Pasteur Institute of Iran, Tehran, Iran.
Iranian Comprehensive Hemophilia Care Center, Tehran, Iran.
Protein Pept Lett. 2023;30(5):401-410. doi: 10.2174/0929866530666230403093538.
Effective T-cell-mediated immunity has emerged as an essential component of human immunodeficiency virus-1 (HIV-1) vaccination. Thus, inducing an immune response against HIV proteins such as Nef and Vif, two major accessory proteins with critical roles in HIV pathogenesis and immune evasion, may lead to an effective approach.
Our goal is to evaluate and compare Montanide ISA-720 and heat shock protein 27 in increasing immunostimulatory properties of HIV-1 Nef-Vif fusion protein as a vaccine candidate.
In this study, the fusion gene with and without the gene was cloned in the prokaryotic pET24a (+) vector. Then, the recombinant Nef-Vif and Hsp27-Nef- Vif proteins were generated in the E. coli system. Finally, their immunostimulatory properties were evaluated in mice. Indeed, the potency of Hsp27 as an endogenous natural adjuvant was investigated to enhance HIV-1 Nef-Vif antigen-specific immunity compared to Montanide ISA-720 as a commercial adjuvant in protein-based immunization strategy.
Our results approved the role of Hsp27 as an effective adjuvant in the stimulation of B- and T-cell immunity. The linkage of Hsp27 to antigen could elicit higher levels of IgG1, IgG2a, IFN-γ, IL- 5 and Granzyme B than antigen mixed with Montanide ISA-720. Moreover, the ratios of IFN-γ/IL-5 and IgG2a/IgG1 were significantly increased in groups receiving Nef-Vif protein + Montanide ISA- 720 and Hsp27-Nef-Vif protein indicating the direction of the immune response pathway toward strong Th1 response. These ratios were higher in the group receiving Hsp27-Nef-Vif protein than in the group receiving Nef-Vif protein + Montanide ISA-720.
Our findings suggest that Hsp27 can be used as an effective adjuvant to enhance antigenspecific immune responses in HIV-1 infectious models for therapeutic vaccine development.
有效的 T 细胞介导的免疫已成为人类免疫缺陷病毒 1（HIV-1）疫苗接种的一个重要组成部分。因此，诱导针对 HIV 蛋白（如 Nef 和 Vif）的免疫反应，这两种主要的辅助蛋白在 HIV 发病机制和免疫逃逸中起着关键作用，可能会带来一种有效的方法。
我们的目标是评估和比较 Montanide ISA-720 和热休克蛋白 27，以提高 HIV-1 Nef-Vif 融合蛋白作为候选疫苗的免疫刺激性。
在这项研究中，带有和不带有 基因的融合基因被克隆到原核 pET24a(+)载体中。然后，在大肠杆菌系统中生成重组 Nef-Vif 和 Hsp27-Nef-Vif 蛋白。最后，在小鼠中评估它们的免疫刺激性。事实上，研究了 Hsp27 作为内源性天然佐剂的效力，以增强 HIV-1 Nef-Vif 抗原特异性免疫，与 Montanide ISA-720 作为蛋白质免疫策略中的商业佐剂相比。
我们的结果证实了 Hsp27 作为 B 细胞和 T 细胞免疫刺激的有效佐剂的作用。与抗原混合的 Hsp27 可以比抗原混合 Montanide ISA-720 产生更高水平的 IgG1、IgG2a、IFN-γ、IL-5 和 Granzyme B。此外，在接受 Nef-Vif 蛋白+Montanide ISA-720 和 Hsp27-Nef-Vif 蛋白的组中，IFN-γ/IL-5 和 IgG2a/IgG1 的比值显著增加，表明免疫反应途径的方向向强烈的 Th1 反应。在接受 Hsp27-Nef-Vif 蛋白的组中，这些比值高于接受 Nef-Vif 蛋白+Montanide ISA-720 的组。
我们的研究结果表明，Hsp27 可用作增强 HIV-1 感染模型中治疗性疫苗开发的抗原特异性免疫反应的有效佐剂。
