Almeida Freya M Freyre, Blanco Aracelys, Trujillo Heidy, Hernández Dunia, García Daymir, Alba José S, Abad Matilde López, Merino Nelson, Lobaina Yadira, Aguilar Rubido Julio C
Vaccine Division, Center for Genetic Engineering and Biotechnology, Havana, Cuba.
Animal Facilities, Center for Genetic Engineering and Biotechnology, Havana, Cuba.
Euroasian J Hepatogastroenterol. 2016 Jan-Jun;6(1):25-30. doi: 10.5005/jp-journals-10018-1161. Epub 2016 Jul 9.
The development of therapeutic vaccines against chronic hepatitis B requires the capacity of the formulation to subvert a tolerated immune response as well as the evaluation of histopathological damage resulting from the treatment. In the present study, the dynamicity of induced immune response to hepatitis B surface antigen (HBsAg) was evaluated in transgenic mice that constitutively express the HBsAg gene (HBsAg-tg mice). After immunization with a vaccine candidate containing both surface (HBsAg) and core (HBcAg) antigens of hepatitis B virus (HBV), the effect of vaccination on clearance of circulating HBsAg and the potential histological alterations were examined. Transgenic (tg) and non-transgenic (Ntg) mice were immunized by intranasal (IN) and subcutaneous (SC) routes simultaneously. A control group received phosphate-buffered saline (PBS) by IN route and aluminum by SC route. Positive responses, at both humoral and cellular levels, were obtained after five immunizations in HBsAg-tg mice. Such responses were delayed and of lower intensity in tg mice, compared to vaccinated Ntg mice. Serum IgG response was characterized by a similar IgG subclass pattern. Even when HBsAg-specific CD8 T cell responses were clearly detectable by gamma-interferon ELISPOT assay, histopathological alterations were not detected in any organ, including the liver and kidneys. Our study demonstrated, that it is possible to subvert the immune tolerance against HBsAg in tg mice, opening a window for new studies to optimize the schedule, dose, and formulation to improve the immune response to the therapeutic vaccine candidate. These results can be considered a safety proof to support clinical developments for the formulation under study.
Freyre FM, Blanco A, Trujillo H, Hernández D, García D, Alba JS, Lopez M, Merino N, Lobaina Y, Aguilar JC. Dynamic of Immune Response induced in Hepatitis B Surface Antigen-transgenic Mice Immunized with a Novel Therapeutic Formulation. Euroasian J Hepato-Gastroenterol 2016;6(1):25-30.
开发针对慢性乙型肝炎的治疗性疫苗需要制剂具备颠覆耐受免疫反应的能力，以及对治疗引起的组织病理学损伤进行评估。在本研究中，在组成性表达乙肝表面抗原（HBsAg）基因的转基因小鼠（HBsAg-tg小鼠）中评估了对乙肝表面抗原（HBsAg）诱导的免疫反应的动态变化。在用包含乙肝病毒（HBV）表面（HBsAg）和核心（HBcAg）抗原的候选疫苗免疫后，检查了疫苗接种对循环HBsAg清除的影响以及潜在的组织学改变。转基因（tg）和非转基因（Ntg）小鼠通过鼻内（IN）和皮下（SC）途径同时免疫。对照组通过IN途径接受磷酸盐缓冲盐水（PBS），通过SC途径接受铝。在对HBsAg-tg小鼠进行五次免疫后，在体液和细胞水平均获得了阳性反应。与接种疫苗的Ntg小鼠相比，tg小鼠中的此类反应延迟且强度较低。血清IgG反应的特征是IgG亚类模式相似。即使通过γ干扰素ELISPOT测定法可清楚检测到HBsAg特异性CD8 T细胞反应，在包括肝脏和肾脏在内的任何器官中均未检测到组织病理学改变。我们的研究表明，有可能在tg小鼠中颠覆对HBsAg的免疫耐受，为新的研究打开了一扇窗，以优化方案、剂量和制剂，从而改善对候选治疗性疫苗的免疫反应。这些结果可被视为支持所研究制剂临床开发的安全性证据。
Freyre FM，Blanco A，Trujillo H，Hernández D，García D，Alba JS，Lopez M，Merino N，Lobaina Y，Aguilar JC。用新型治疗制剂免疫的乙肝表面抗原转基因小鼠诱导的免疫反应动态。《欧亚肝脏胃肠病学杂志》2016；6（1）：25 - 30。
