Comparison of Adjuvant Effects of Montanide ISA-720 and Heat Shock Protein 27 in Increasing Immunostimulatory Properties of HIV-1 Nef-Vif Fusion Protein Construct.

IF 1 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY
Niloofar Khairkhah, Fatemeh Shahhosseini, Elnaz Agi, Alireza Milani, Azam Bolhassani
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引用次数: 1

Abstract

Introduction: 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.

Aim: 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.

Methods: In this study, the nef-vif fusion gene with and without the heat shock protein 27 (hsp27) 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.

Results: 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.

Conclusion: 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.

Montanide ISA-720和热休克蛋白27在增强HIV-1 Nef-Vif融合蛋白构建体免疫刺激特性中的佐剂作用比较
有效的t细胞介导免疫已成为人类免疫缺陷病毒-1 (HIV-1)疫苗接种的重要组成部分。因此,诱导针对HIV蛋白(如Nef和Vif)的免疫应答可能是一种有效的方法。Nef和Vif是在HIV发病机制和免疫逃避中起关键作用的两种主要辅助蛋白。目的:我们的目标是评估和比较Montanide ISA-720和热休克蛋白27在增加HIV-1 Nef-Vif融合蛋白作为候选疫苗的免疫刺激特性。方法:在原核pET24a(+)载体上克隆含热休克蛋白27 (hsp27)基因和不含hsp27基因的nef-vif融合基因。然后,在大肠杆菌系统中生成重组Nef-Vif和Hsp27-Nef- Vif蛋白。最后,在小鼠中评价其免疫刺激特性。事实上,与Montanide ISA-720作为商业佐剂相比,研究人员研究了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。结果:我们的研究结果证实了Hsp27作为一种有效的佐剂在刺激B细胞和t细胞免疫中的作用。与与Montanide ISA-720混合的抗原相比,Hsp27与抗原的连锁反应可引起更高水平的IgG1、IgG2a、IFN-γ、IL- 5和颗粒酶B。Nef-Vif蛋白+ Montanide ISA- 720和Hsp27-Nef-Vif蛋白组IFN-γ/IL-5和IgG2a/IgG1比值显著升高,提示免疫应答途径向强Th1应答方向发展。这些比率在接受Hsp27-Nef-Vif蛋白的组高于接受Nef-Vif蛋白+ Montanide ISA-720的组。结论:我们的研究结果表明,Hsp27可以作为一种有效的佐剂,增强HIV-1感染模型中的抗原特异性免疫反应,用于治疗性疫苗的开发。
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来源期刊
Protein and Peptide Letters
Protein and Peptide Letters 生物-生化与分子生物学
CiteScore
2.90
自引率
0.00%
发文量
98
审稿时长
2 months
期刊介绍: Protein & Peptide Letters publishes letters, original research papers, mini-reviews and guest edited issues in all important aspects of protein and peptide research, including structural studies, advances in recombinant expression, function, synthesis, enzymology, immunology, molecular modeling, and drug design. Manuscripts must have a significant element of novelty, timeliness and urgency that merit rapid publication. Reports of crystallization and preliminary structure determination of biologically important proteins are considered only if they include significant new approaches or deal with proteins of immediate importance, and preliminary structure determinations of biologically important proteins. Purely theoretical/review papers should provide new insight into the principles of protein/peptide structure and function. Manuscripts describing computational work should include some experimental data to provide confirmation of the results of calculations. Protein & Peptide Letters focuses on: Structure Studies Advances in Recombinant Expression Drug Design Chemical Synthesis Function Pharmacology Enzymology Conformational Analysis Immunology Biotechnology Protein Engineering Protein Folding Sequencing Molecular Recognition Purification and Analysis
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