Gholami Parizad Elaheh, Imani Fooladi Abbas Ali, Sedighian Hamid, Behzadi Elham, Valizadeh Azar, Khosravi Afra
Clinical Microbiology Research Center, Ilam University of Medical Sciences, Ilam, Iran.
Applied Microbiology Research Center, Systems Biology and Poisonings Institute, Baqiyatallah University of Medical Sciences, Tehran, Iran.
Iran J Pathol. 2022 Fall;17(4):448-460. doi: 10.30699/IJP.2022.553785.2896. Epub 2022 Sep 2.
BACKGROUND & OBJECTIVE: The vaccine available to prevent Hepatitis B virus disease is ineffective in 5% of people due to the use of HBsAg as a weak immunogenic factor. In the present study, PreS2/S fused to C18-27 peptide fragment as an effective antigen and is proposed as a promising vaccine candidate compared with the conventional vaccine prescribed in the vaccination program.
After the synthesis of PreS2/S genes and C18-27 peptide fragment in pET28a, the recombinant protein was confirmed by Western blotting. The efficacy of the PreS2/S-C18-27 protein was compared with the conventional vaccine injected into five groups of rats. Finally, the cytokine level of IF-r, IL-2, IL-4, IL-10, TNF-a, IgG1, and IgG2a were measured using the ELISA method.
This study showed no significant difference between the recombinant vaccine group and PBS control group in the IF-r test, but there was a significant difference between groups testing IL-2 and IL-10. In addition, the group receiving the recombinant vaccine with CPG adjuvant at a dilution of 1/10 in the IgG total test on days 14 and 45 after the first injection showed a significant difference in comparison with other groups.
This study showed no statistically significant difference between the recombinant protein vaccine group and the conventional vaccine group. The Th1- mediated immune responses obtained from recombinant proteins with and without CPG performed better than conventional vaccines, possibly due to the functional deficiency of the available vaccines.
由于将乙肝表面抗原（HBsAg）用作弱免疫原性因子，现有的预防乙型肝炎病毒疾病的疫苗在5%的人群中无效。在本研究中，PreS2/S与C18 - 27肽片段融合作为一种有效的抗原，与疫苗接种计划中规定的传统疫苗相比，被提议作为一种有前景的候选疫苗。
在pET28a中合成PreS2/S基因和C18 - 27肽片段后，通过蛋白质印迹法确认重组蛋白。将PreS2/S - C18 - 27蛋白的效力与注射到五组大鼠体内的传统疫苗进行比较。最后，使用酶联免疫吸附测定法（ELISA）测量干扰素-γ（IF-γ）、白细胞介素-2（IL-2）、白细胞介素-4（IL-4）、白细胞介素-10（IL-10）、肿瘤坏死因子-α（TNF-α）、免疫球蛋白G1（IgG1）和免疫球蛋白G2a（IgG2a）的细胞因子水平。
本研究表明，在IF-γ检测中，重组疫苗组与磷酸盐缓冲盐水（PBS）对照组之间无显著差异，但在检测IL-2和IL-10的组间存在显著差异。此外，在首次注射后第14天和第45天的IgG总量检测中，接受1/10稀释度含胞嘧啶-磷酸-鸟嘌呤（CPG）佐剂的重组疫苗组与其他组相比存在显著差异。
本研究表明重组蛋白疫苗组与传统疫苗组之间无统计学显著差异。从含和不含CPG的重组蛋白获得的Th1介导的免疫反应比传统疫苗表现更好，这可能是由于现有疫苗的功能缺陷。
