Sadat Larijani Mona, Ramezani Amitis, Mashhadi Abolghasem Shirazi Maryam, Bolhassani Azam, Pouriayevali Mohammad Hassan, Shahbazi Sepideh, Sadat Seyed Mehdi
Clinical Research Department, Pasteur Institute of Iran, Tehran, Iran; Hepatitis, AIDS and Blood borne diseases Department, Pasteur Institute of Iran, Tehran, Iran.
Clinical Research Department, Pasteur Institute of Iran, Tehran, Iran.
Virus Res. 2021 Jun;298:198403. doi: 10.1016/j.virusres.2021.198403. Epub 2021 Mar 26.
Various approaches have been investigated to prevent or eliminate HIV-1 since 1981. However, the virus has been affecting human population worldwide with no effective vaccine yet. The conserved regions among the viral genes are suitable targets in mutable viruses to induce the immune responses via an effective delivery platform. In this study, we aimed at evaluation of p24 and nef in two forms of full and truncated genes as two fusion antigenic forms according to our previous bioinformatics analysis. The designed antigens were then transferred through ex vivo generated dendritic cells and also proteins in BALB/c to assess and compare immunogenicity. p24 and Nef amino acid sequences were aligned, then, the most conserved regions were selected and two fusion forms as the truncated (p24:80-231aa-Nef:120-150aa) and the full from (p24-Nef) were cloned and expressed in prokaryotic and eukaryotic systems. Lentiviral vectors were applied to generate recombinant virions harboring the genes of interest to transduce generated murine dendritic cells. BALB/c mice received the recombinant DCs or recombinant proteins according to the defined schedule. IgG development was assessed to determine humoral immune activity and cellular immune responses were evaluated by IL-5 and IFN-y induction. Granzyme B secretion was also investigated to determine CTL activity in different immunized groups. The data showed high induction of cellular immune responses in dendritic cell immunization specifically in immunized mice with the truncated form of the p24 and Nef by high secretion of IFN-y and strong CTL activity. Moreover, protein/ DC prime-boost formulation led to stronger Th1 pathway and strong CTL activation in comparison with other formulations. The generated recombinant dendritic cells expressing p24-Nef induced humoral and cellular immunity in a Th1 pathway specifically with the in silico predicted truncated antigen which could be of high value as a dendritic cell therapeutic vaccine candidate against HIV-1.
自1981年以来，人们研究了各种预防或消除HIV-1的方法。然而，这种病毒一直在全球范围内影响着人类，至今仍没有有效的疫苗。病毒基因中的保守区域是可变病毒中合适的靶点，可通过有效的递送平台诱导免疫反应。在本研究中，根据我们之前的生物信息学分析，我们旨在评估两种形式（全长和截短）的p24和nef作为两种融合抗原形式。然后将设计的抗原通过体外产生的树突状细胞以及BALB/c小鼠体内的蛋白质进行传递，以评估和比较免疫原性。对p24和Nef氨基酸序列进行比对，然后选择最保守的区域，克隆截短形式（p24:80-231aa-Nef:120-150aa）和全长形式（p24-Nef）这两种融合形式，并在原核和真核系统中表达。应用慢病毒载体产生携带感兴趣基因的重组病毒颗粒，以转导产生的小鼠树突状细胞。BALB/c小鼠按照规定的时间表接受重组树突状细胞或重组蛋白。评估IgG的产生以确定体液免疫活性，并通过IL-5和IFN-γ的诱导来评估细胞免疫反应。还研究了颗粒酶B的分泌，以确定不同免疫组中的CTL活性。数据显示，树突状细胞免疫可高度诱导细胞免疫反应，特别是在用截短形式的p24和Nef免疫的小鼠中，IFN-γ分泌量高且CTL活性强。此外，与其他制剂相比，蛋白质/树突状细胞初免-加强制剂导致更强的Th1途径和更强的CTL激活。所产生的表达p24-Nef的重组树突状细胞在Th1途径中诱导了体液和细胞免疫，特别是与计算机预测的截短抗原一起，这作为一种抗HIV-1的树突状细胞治疗疫苗候选物可能具有很高的价值。
