Research and Development, SEEK, 45 Beech Street, London, EC2Y 8AD, UK.
Virol J. 2013 Apr 4;10:107. doi: 10.1186/1743-422X-10-107.
This manuscript describes the development of a novel synthetic immunotherapy (HIV-v) composed of four multi-epitope polypeptides targeting conserved regions in the Nef, Rev, Vif and Vpr viral proteins. Immunogenicity and cytotoxicity of HIV-v are discussed.
Short conserved T-cell multi-epitope regions were identified in silico in the HIV proteome. The immunogenicity of the identified HIV-v polypeptides was assessed in vivo by immunisation of C57BLK6 mice transgenic for HLA-A*0201. Splenocytes from immunised animals were exposed in vitro to soluble HIV-v polypeptides or to syngeneic (T1) or allogeneic (Jurkat) cells transfected with these polypeptides. Specific T-cell reactivity was assessed by cell-based IFN-γ ELISA. Virus specific CD3 + CD8+ IFN-γ+ recall responses were also determined by flow cytometry following in vitro exposure of splenocytes from immunised mice to syngeneic (T1) and allogeneic (H9) cells infected with HIV-1 strain IIIB. HIV-v specific antibodies were quantified by ELISA whilst antibody mediated anti-viral immunotherapeutic effect on T1 cells infected with a laboratory adapted and a primary isolate of the HIV-1 virus was assessed in a LDH-based complement mediated lysis assay.
HIV-v elicited antigen-specific IgG and IFN-γ responses against the synthetic polypeptides in the formulation. HIV-v specific T cells recognised polypeptides presented either as soluble antigen or complexed to HLA-A*0201 following natural processing and presentation by syngeneic human T1 cells. Moreover, the CD3 + CD8+ component of the response recognised syngeneic T1 cells naturally infected with HIV-1 in a virus-specific and MHC restricted-manner. The HIV-v specific IgG response was also able to recognise human T1 cells naturally infected with HIV-1 and induce cell death through classic activation of complement.
HIV-v induces a vaccine-specific type I immune response characterised by activation of effector CD8+ T cell and antibody responses that recognise and kill human cell lines naturally infected with a laboratory adapted and a primary isolate of the HIV-1 virus. The data supports the hypothesis that alternative HIV protein targets can be effectively used to prime both cellular and antibody immune responses of clinical value in the prevention and treatment of HIV infection.
本文描述了一种新型合成免疫疗法（HIV-v）的开发，该疗法由针对 HIV 病毒蛋白 Nef、Rev、Vif 和 Vpr 中保守区域的四个多表位多肽组成。讨论了 HIV-v 的免疫原性和细胞毒性。
通过对 HLA-A*0201 转基因 C57BLK6 小鼠进行免疫接种，在体内评估鉴定的 HIV-v 多肽的免疫原性。从免疫动物的脾细胞在体外暴露于可溶性 HIV-v 多肽或用这些多肽转染的同基因（T1）或同种异体（Jurkat）细胞。通过细胞基础 IFN-γ ELISA 评估特异性 T 细胞反应。通过流式细胞术也确定了来自免疫小鼠的脾细胞在体外暴露于感染 HIV-1 株 IIIB 的同基因（T1）和同种异体（H9）细胞后病毒特异性 CD3+CD8+IFN-γ+回忆反应。通过 ELISA 定量 HIV-v 特异性抗体，通过 LDH 基于补体介导的溶解测定评估针对感染实验室适应和原发性 HIV-1 病毒的 T1 细胞的抗体介导抗病毒免疫治疗效果。
HIV-v 在制剂中引发针对合成多肽的抗原特异性 IgG 和 IFN-γ 反应。HIV-v 特异性 T 细胞识别作为可溶性抗原或与 HLA-A*0201 复合的多肽，通过同基因人 T1 细胞的天然加工和呈递。此外，该反应的 CD3+CD8+成分以病毒特异性和 MHC 限制方式识别自然感染 HIV-1 的同基因 T1 细胞。HIV-v 特异性 IgG 反应还能够识别自然感染 HIV-1 的人 T1 细胞，并通过经典激活补体诱导细胞死亡。
HIV-v 诱导一种疫苗特异性 I 型免疫反应，其特征在于激活效应 CD8+T 细胞和抗体反应，该反应识别并杀伤自然感染实验室适应和原发性 HIV-1 病毒的人细胞系。数据支持替代 HIV 蛋白靶标可有效用于启动细胞和抗体免疫反应的假说，这些反应具有临床价值，可用于预防和治疗 HIV 感染。
