Sadeghi Sahar, Bandehpour Mojgan, Haji Molla Hoseini Mostafa, Sharifnia Zarin
Department of Medical Biotechnology, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
Cellular and Molecular Biology Research Center, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
Iran J Basic Med Sci. 2021 Aug;24(8):1126-1137. doi: 10.22038/IJBMS.2021.58087.12909.
Prevalence of virus, creates the need to achieve an efficient vaccine against it. We examined whether the predicted antigenic epitopes of HA, NP, and M2 proteins of the influenza H1N1 and H3N2 viruses accompanied by chitin and chitosan biopolymers might be relevant to the induction of effective proper mucosal responses.
The construct was prepared using B and T cell predicted epitopes of HA, NP, and M2 proteins from the influenza H1N1 and H3N2 viruses by considering haplotype "d" as a dominant allele in the BALB/c mice. Intranasal immunization with purified LPS free recombinant protein together with chitin and chitosan microparticles as adjuvants was administered at an interval of 2 weeks in thirty-five BALB/c female mice which were divided into seven groups. Ten days after the last immunization, humoral and cellular immune responses were examined.
Elevated systemic IgG2a, IgA, and mucosal IgA revealed a humoral response to the construct. An increase in the number of IFN-γ-producing cells in re-stimulation of splenocytes in the culture medium by poly-tope as well as rise in the concentrations of IL-6, IL-17, and TNF-α along with the regulatory response of IL-10, presented the capacity of the designed protein to provoke significant immune responses. The neutralization test ultimately confirmed the high efficacy of the protein in inhibiting the virus.
The results support the fact that immunogenic poly-tope protein in the presence of chitin and chitosan microparticles as mucosal adjuvants is able to induce humoral and cell-mediated responses in BALB/c mice.
病毒的流行，使得研发一种有效的疫苗成为必要。我们研究了流感H1N1和H3N2病毒的血凝素（HA）、核蛋白（NP）和M2蛋白的预测抗原表位，以及几丁质和壳聚糖生物聚合物是否可能与诱导有效的适当黏膜反应相关。
构建体是通过考虑BALB/c小鼠中作为优势等位基因的单倍型“d”，利用流感H1N1和H3N2病毒HA、NP和M2蛋白的B细胞和T细胞预测表位制备而成。将纯化的无脂多糖重组蛋白与几丁质和壳聚糖微粒作为佐剂进行鼻内免疫，以2周的间隔对35只BALB/c雌性小鼠给药，这些小鼠被分为7组。最后一次免疫后10天，检测体液免疫和细胞免疫反应。
全身IgG2a、IgA以及黏膜IgA升高表明对构建体有体液免疫反应。通过多表位在培养基中再次刺激脾细胞时，产生γ干扰素的细胞数量增加，以及IL-6、IL-17和TNF-α浓度升高，同时伴有IL-10的调节反应，表明设计的蛋白有引发显著免疫反应的能力。中和试验最终证实了该蛋白在抑制病毒方面的高效性。
结果支持了这样一个事实，即在几丁质和壳聚糖微粒作为黏膜佐剂存在的情况下，免疫原性多表位蛋白能够在BALB/c小鼠中诱导体液免疫和细胞介导的反应。
