Department of Clinical Laboratory, The First Affiliated Hospital of Xinjiang Medical University, Urumqi 830011, Xinjiang, PR China.
Department of Clinical Laboratory, The Third Affiliated Hospital of Xinjiang Medical University, Urumqi 830011, Xinjiang, PR China.
Mol Immunol. 2018 Feb;94:183-189. doi: 10.1016/j.molimm.2018.01.002. Epub 2018 Jan 10.
This study is to predict and purify the T-B combined epitopes of egG1Y162 antigen in Echinococcus granulosus, and to evaluate their immunogenicity in mice.
The bioinformatics software was used to predict the T-B combined epitopes of egG1Y162 antigen. Recombinant egG1Y161/2 peptides were constructed, expressed and purified. Mice were immunized with egG1Y161/2 peptides. The serum and spleen cells were isolated. The isolated spleen cells were stimulated with egG1Y161/2 peptides in vitro and the culture supernatant was collected. The levels of IgG in serum and levels of IL-4 and IFN-γ in the culture supernatant were measured by ELISA. The weight and number of the fresh hydatid cysts were evaluated. The serum ptotoscolicidal activity was measured by the complement dependent cytotoxicity assay.
Peptides of 6-19aa, 64-82aa, 106-119aa were predicted as T-B combined epitopes of egG1Y162 antigen. And, recombinant protein egG1Y162-1 or egG1Y162-2, which contained T-B combined epitope(s) of the 6-19aa, or the 64-82aa and the 106-119aa in egG1Y162 antigen, respectively, was successfully expressed and purified. Serum IgG levels of mice immunized with egG1Y162-1/2 were significantly increased during the immune response to Echinococcus granulosus. The levels of IFN-γ, IL-4 and the ratio of IFN-γ/IL-4 after egG1Y162-1/2 immunization were significantly higher. Weight and number of the fresh hydatid cysts in egG1Y162-1/2 immunized mice was significantly decreased. And, the serum protoscolicidal activity after egG1Y162-1/2 immunization was enhanced.
The egG1Y162-1/2 induces production of serum IgG levels and Th1 cell immune response, which enhances the protective immunity in Echinococcus granulosus challenged mice and thus may be used as a potential vaccine candidate.
本研究旨在预测并纯化细粒棘球绦虫 egG1Y162 抗原的 T-B 联合表位，并评估其在小鼠中的免疫原性。
使用生物信息学软件预测 egG1Y162 抗原的 T-B 联合表位。构建、表达和纯化重组 egG1Y161/2 肽。用 egG1Y161/2 肽免疫小鼠，分离血清和脾细胞。分离的脾细胞用 egG1Y161/2 肽体外刺激，收集培养上清液。ELISA 法检测血清 IgG 水平和培养上清液中 IL-4 和 IFN-γ 水平。评估新鲜包虫囊肿的重量和数量。用补体依赖性细胞毒性试验测定血清原虫杀伤活性。
预测 egG1Y162 抗原的 6-19aa、64-82aa、106-119aa 肽段为 T-B 联合表位。成功表达并纯化了含有 egG1Y162 抗原 6-19aa、64-82aa 和 106-119aa 中 T-B 联合表位之一的重组蛋白 egG1Y162-1 或 egG1Y162-2。egG1Y162-1/2 免疫后，小鼠对细粒棘球蚴的免疫反应中血清 IgG 水平显著升高。egG1Y162-1/2 免疫后 IFN-γ、IL-4 水平及 IFN-γ/IL-4 比值显著升高。egG1Y162-1/2 免疫小鼠的新鲜包虫囊肿重量和数量显著减少。egG1Y162-1/2 免疫后血清原虫杀伤活性增强。
egG1Y162-1/2 诱导产生血清 IgG 水平和 Th1 细胞免疫应答，增强了细粒棘球蚴感染小鼠的保护性免疫，因此可能成为一种有潜力的疫苗候选物。
