Mishra Sasmita, Lavelle Bianca J, Desrosiers Joe, Ardito Matt T, Terry Frances, Martin William D, De Groot Anne S, Gregory Stephen H
Department of Medicine, Rhode Island Hospital and the Warren Alpert Medical School of Brown University, Providence, Rhode Island, United States of America.
Institute for Immunology and Informatics, University of Rhode Island, Providence, Rhode Island, United States of America.
PLoS One. 2014 Aug 11;9(8):e104606. doi: 10.1371/journal.pone.0104606. eCollection 2014.
Hepatitis C virus (HCV) is the etiologic agent of chronic liver disease, hepatitis C. Spontaneous resolution of viral infection is associated with vigorous HLA class I- and class II-restricted T cell responses to multiple viral epitopes. Unfortunately, only 20% of patients clear infection spontaneously, most develop chronic disease and require therapy. The response to chemotherapy varies, however; therapeutic vaccination offers an additional treatment strategy. To date, therapeutic vaccines have demonstrated only limited success. Vector-mediated vaccination with multi-epitope-expressing DNA constructs alone or in combination with chemotherapy offers an additional treatment approach. Gene sequences encoding validated HLA-A2- and HLA-DRB1-restricted epitopes were synthesized and cloned into an expression vector. Dendritic cells (DCs) derived from humanized, HLA-A2/DRB1 transgenic (donor) mice were transfected with these multi-epitope-expressing DNA constructs. Recipient HLA-A2/DRB1 mice were vaccinated s.c. with transfected DCs; control mice received non-transfected DCs. Peptide-specific IFN-γ production by splenic T cells obtained at 5 weeks post-immunization was quantified by ELISpot assay; additionally, the production of IL-4, IL-10 and TNF-α were quantified by cytokine bead array. Splenocytes derived from vaccinated HLA-A2/DRB1 transgenic mice exhibited peptide-specific cytokine production to the vast majority of the vaccine-encoded HLA class I- and class II-restricted T cell epitopes. A multi-epitope-based HCV vaccine that targets DCs offers an effective approach to inducing a broad immune response and viral clearance in chronic, HCV-infected patients.
丙型肝炎病毒(HCV)是慢性肝病丙型肝炎的病原体。病毒感染的自发清除与针对多个病毒表位的强烈的HLA I类和II类限制性T细胞反应相关。不幸的是,只有20%的患者能自发清除感染,大多数患者会发展为慢性病并需要治疗。然而,对化疗的反应各不相同;治疗性疫苗提供了另一种治疗策略。迄今为止,治疗性疫苗仅取得了有限的成功。单独或与化疗联合使用表达多表位的DNA构建体进行载体介导的疫苗接种提供了另一种治疗方法。合成了编码经过验证的HLA-A2和HLA-DRB1限制性表位的基因序列,并将其克隆到一个表达载体中。用这些表达多表位的DNA构建体转染源自人源化、HLA-A2/DRB1转基因(供体)小鼠的树突状细胞(DC)。受体HLA-A2/DRB1小鼠经皮下接种转染的DC;对照小鼠接受未转染的DC。通过ELISpot测定法对免疫后5周获得的脾T细胞产生的肽特异性IFN-γ进行定量;此外,通过细胞因子珠阵列对IL-4、IL-10和TNF-α的产生进行定量。源自接种疫苗的HLA-A2/DRB1转基因小鼠的脾细胞对绝大多数疫苗编码的HLA I类和II类限制性T细胞表位表现出肽特异性细胞因子产生。一种靶向DC的基于多表位的HCV疫苗为在慢性HCV感染患者中诱导广泛的免疫反应和病毒清除提供了一种有效方法。