Innate & adaptive immunities in control of ocular HSV

先天

• 批准号: 6866261
• 负责人: HOMAYON GHIASI
• 金额: $ 35.1万
• 依托单位: CEDARS-SINAI MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-02-01 至 2008-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6866261
• 关键词: Herpesviridae vaccine; active immunization; cytotoxic T lymphocyte; glycoproteins; herpes simplex virus 1; immunity; interferon gamma; interleukin 12; laboratory mouse; natural killer cells; ocular herpes; vaccine development; vaccine evaluation; virus protein

DESCRIPTION (provided by applicant): Following primary herpes simplex virus type-1 (HSV-1) infection of the eye, virus replicates in the eye. Eye disease can result from direct damage inflicted on the corneal tissue by HSV-1, or indirectly, due to cytokines and chemokines released from cells infiltrating the cornea after infection. Virus is completely cleared between days 7-10 following ocular infection. The longer it takes for the immune system to clear the virus from the eye; the more extensive and protracted is the consequent ocular disease. Decreased virus load in the eye and accelerated virus clearance are probably the most efficient way to decrease latency and thus subsequent recurrent infections and loss of vision. Therefore, our main goal is to determine immune responses that are most efficacious at decreasing HSV-1 replication in the eye, reducing the establishment of latency, and reactivation from latency. We have shown recently that: (1) Immunization with a cocktail of 5 HSV-1 glycoproteins (5gP) (gB, gC, gD, gE, gl) is more efficacious than immunization with any single glycoprotein, gB+gD, or live HSV-1 vaccine; and (2) Immunization with 5gP DNA ("naked" DNA corresponding to the 5gP proteins) appeared even more efficacious than 5gP protein (subunit) immunization, resulting in further reduction of virus titers in the eye, more rapid viral clearance, lower establishment of latency in trigeminal ganglia (TGs), higher IL-12, and higher IFN-gamma (Appendix 1). Most recently, we have shown that DNA immunization also stimulates NK cells, macrophages and produces significantly higher numbers of CD11c+ and CD8 + T cells in the cornea of DNA-immunized mice than protein-immunized mice. We hypothesize that following DNA immunization signaling through dendritic cells (DCs) is important to the enhancement and maintenance of the production of CD8+/NK cells in the cornea of DNA-immunized mice. We further hypothesize that following DNA immunization signaling through macrophages (IL-12 production) is important in to the provision of the CD8+/NK cells with the signaling required to enhance and maintain IFN-gamma, production. Thus, to further elucidate the biologic and immunologic mechanisms responsible for reduced virus replication in the eye, more rapid clearance of HSV-1, and reduce latency in mice immunized with 5gP DNA, we propose to accomplish the following Specific Aims: 1. Confirm the hypothesis that the higher efficacy of 5gP DNA immunization is due to stimulation of both innate and adaptive immune responses, whereas the relatively lower efficacy of 5gP protein (the most effective protein vaccine against ocular HSV-1) immunization is mostly due to CD4-related responses. 2. Confirm the hypothesis that further stimulation of professional APCs using IL-12 and FLt3L as adjuvants will further enhance the activities of CD8 + T cells and NK cells leading to a greater reduction in ocular virus replication, latency, and reactivation in DNA-vaccinated mice.

描述（由申请人提供）：眼睛原发性单纯疱疹病毒 1 型 (HSV-1) 感染后，病毒在眼睛中复制。眼部疾病可能是由于 HSV-1 对角膜组织造成的直接损害造成的，也可能是感染后浸润角膜的细胞释放的细胞因子和趋化因子造成的间接损害。眼部感染后 7-10 天，病毒被完全清除。免疫系统清除眼睛中的病毒所需的时间越长；随之而来的眼部疾病更加广泛和持久。减少眼睛中的病毒载量和加速病毒清除可能是减少潜伏期以及随后的复发性感染和视力丧失的最有效方法。因此，我们的主要目标是确定在减少眼内 HSV-1 复制、减少潜伏期的建立和从潜伏期重新激活方面最有效的免疫反应。我们最近证明：（1）使用 5 种 HSV-1 糖蛋白（5gP）（gB、gC、gD、gE、gl）的混合物进行免疫比使用任何单一糖蛋白、gB+gD 或 HSV-1 活疫苗进行免疫更有效； (2) 5gP DNA（对应于 5gP 蛋白的“裸露”DNA）免疫似乎比 5gP 蛋白（亚基）免疫更有效，导致眼内病毒滴度进一步降低、病毒清除更快、三叉神经节 (TG) 潜伏期缩短、IL-12 升高和 IFN-γ 升高（附录 1）。最近，我们发现 DNA 免疫还可以刺激 NK 细胞、巨噬细胞，并在 DNA 免疫小鼠的角膜中产生比蛋白质免疫小鼠显着更多数量的 CD11c+ 和 CD8 + T 细胞。我们假设，通过树突状细胞 (DC) 传递 DNA 免疫信号对于增强和维持 DNA 免疫小鼠角膜中 CD8+/NK 细胞的产生非常重要。我们进一步假设，通过巨噬细胞的 DNA 免疫信号传导（IL-12 产生）对于向 CD8+/NK 细胞提供增强和维持 IFN-γ 产生所需的信号传导非常重要。因此，为了进一步阐明导致眼睛中病毒复制减少、HSV-1 更快清除以及减少 5gP DNA 免疫小鼠潜伏期的生物学和免疫机制，我们建议实现以下具体目标： 1. 证实5gP DNA免疫的较高功效是由于刺激 先天性和适应性免疫反应，而 5gP 蛋白（针对眼部 HSV-1 最有效的蛋白疫苗）免疫效果相对较低，主要是由于 CD4 相关反应。 2. 证实使用 IL-12 和 FLt3L 进一步刺激专业 APC 的假设 佐剂将进一步增强 CD8 + T 细胞和 NK 细胞的活性，从而更大程度地减少 DNA 疫苗接种小鼠眼部病毒的复制、潜伏和重新激活。