Trujillo H, Blanco A, García D, Freyre F, Aguiar J, Lobaina Y, Aguilar J C
Hepatitis B Department, Biomedical Research Unit, Center for Genetic Engineering and Biotechnology, Havana, Cuba.
Animal Facilities, Center for Genetic Engineering and Biotechnology, Havana, Cuba.
Euroasian J Hepatogastroenterol. 2014 Jul-Dec;4(2):70-78. doi: 10.5005/jp-journals-10018-1105. Epub 2014 Jul 28.
Hepatitis B core antigens (HBcAg) and hepatitis B surface antigens (HBsAg) are the main structural antigens of hepatitis B virus (HBV). Both antigens are potent immunogens for experimental animals as well as in acutely infected patients. A novel formulation based on the combination of HBsAg and HBcAg has been developed as a therapeutic vaccine candidate, aimed at inducing an immune response capable of controlling the infection. An immunization schedule was conducted to evaluate the immunogenicity of this formulation after simultaneous immunization by the intranasal and parenteral routes using different schedules and doses. Humoral and cellular immune responses generated in blood and spleen were evaluated by engyme-linked immunosorbent assay (ELISA) and enzyme-liked immunospot (ELISPOT) assays respectively. A first experiment evaluated two groups of mice simultaneously immunized by intranasal (IN) and subcutaneous (SC) routes, one including alum by SC route and, in the other, the formulation was injected without adjuvant. As a result, alum adjuvant did not increase the immunogenicity under the studied conditions. In fact, the group without alum induced the most potent immune response. The immune response was enhanced by combining IN and SC immunization compared to the SC route alone. In a second experiment, mice were immunized by different mucosal routes at the same time, and compared to the simultaneously (IN/SC) immunized groups. It was demonstrated that there is no improvement on the resulting immune response by using multiple routes of immunizations simultaneously; however, the increase of the antigen dose induced a superior immune response. Interestingly, the increase of antigen dose only by SC route did not favor the resulting immunogenicity. In conclusion, the use of HBsAg transgenic mice has proven useful to optimize the formulation, avoiding the unnecessary use of alum as adjuvant as well as provided information of the role of different mucosal immunization routes and antigen dose on the resulting immune response. Trujillo H, Blanco A, García D, Freyre F, Aguiar J, Lobaina Y, Aguilar JC. Optimization of a Therapeutic Vaccine Candidate by Studying Routes, Immunization Schedules and Antigen Doses in HBsAg-positive Transgenic Mice. Euroasian J Hepato-Gastroenterol 2014;4(2):70-78.
乙肝核心抗原（HBcAg）和乙肝表面抗原（HBsAg）是乙肝病毒（HBV）的主要结构抗原。这两种抗原对于实验动物以及急性感染患者而言都是强效免疫原。一种基于HBsAg和HBcAg组合的新型制剂已被开发出来作为治疗性疫苗候选物，旨在诱导能够控制感染的免疫反应。通过使用不同的方案和剂量，经鼻内和肠胃外途径同时免疫后，实施了一个免疫程序来评估该制剂的免疫原性。分别通过酶联免疫吸附测定（ELISA）和酶联免疫斑点（ELISPOT）测定来评估血液和脾脏中产生的体液免疫和细胞免疫反应。第一个实验评估了两组经鼻内（IN）和皮下（SC）途径同时免疫的小鼠，一组通过皮下途径使用明矾，另一组则在不使用佐剂的情况下注射该制剂。结果，在研究条件下明矾佐剂并未增加免疫原性。事实上，未使用明矾的组诱导出了最有效的免疫反应。与单独的皮下途径相比,经鼻内和皮下联合免疫增强了免疫反应。在第二个实验中，小鼠同时通过不同的黏膜途径进行免疫，并与同时（经鼻内/皮下）免疫的组进行比较。结果表明，同时使用多种免疫途径并不会改善所产生的免疫反应；然而，抗原剂量的增加诱导出了更强的免疫反应。有趣的是，仅通过皮下途径增加抗原剂量对所产生的免疫原性并无益处。总之，已证明使用HBsAg转基因小鼠有助于优化制剂，避免不必要地使用明矾作为佐剂，并提供了不同黏膜免疫途径和抗原剂量对所产生免疫反应的作用的信息。 特鲁希略H，布兰科A，加西亚D，弗雷雷F，阿吉亚尔J，洛巴伊纳Y，阿吉拉尔JC。通过研究HBsAg阳性转基因小鼠的免疫途径、免疫程序和抗原剂量优化治疗性疫苗候选物。《欧亚肝脏胃肠病学杂志》2014年；4(2):70 - 78。
