A Novel Prime/Pull Therapeutic Vaccine Strategy to Prevent Recurrent Genital Herpes

预防复发性生殖器疱疹的新型初免/拉动治疗疫苗策略

• 批准号: 9913971
• 负责人: Lbachir BenMohamed
• 金额: $ 69.75万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-01-10 至 2024-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9913971
• 关键词: Adult; Affect; Afferent Neurons; Age-Years; Animal Model; Antibodies; Antibody Response; Antigens; Antiviral Agents; Axon; B-Lymphocytes; Back; CD4 Positive T Lymphocytes; CD8-Positive T-Lymphocytes; CXCL10 gene; CXCL11 gene; CXCL9 gene; CXCR3 gene; Cavia; Cessation of life; Clinic; Clinical Trials; Data; Disease; Disease Outbreaks; Epithelial; Epithelium; FDA approved; Foundations; Goals; HIV; Herpesviridae Infections; Human; Human Herpesvirus 2; Immunity; Infection; Knowledge; Lead; Lesion; Ligands; Medical; Memory; Mission; Modeling; National Institute of Allergy and Infectious Disease; Neurons; Newborn Infant; Peripheral; Pharmacotherapy; Phenotype; Plant Roots; Prevalence; Primary Infection; Protein Subunits; Proteins; Publishing; Recurrence; Ribonucleotide Reductase Subunit; Risk; Safety; Scientist; Simplexvirus; Site; Spinal Ganglia; Subunit Vaccines; Surface; Symptoms; System; T cell response; T memory cell; T-Lymphocyte; Testing; Therapeutic; Tissues; Travel; United States; Ursidae Family; VP 16; Vaccine Research; Vaccine Therapy; Vaccines; Vagina; Virion; Virus; Virus Latency; Virus Replication; Woman; adeno-associated viral vector; base; chemokine; design; genital herpes; healing; human disease; improved; innovation; insight; latent infection; men; multidisciplinary; nanoparticle; neonate; nervous system disorder; neurotropic; novel; pre-clinical; preclinical study; prevent; prophylactic; protective efficacy; psychologic; reactivation from latency; reproductive tract; seropositive; therapeutic vaccine; transcriptome sequencing; transcriptomics; vaginal infection; vector; viral transmission

PROJECT SUMMARY/ABSTRACT Genital herpes simplex type virus-2 (HSV-2) infection affects over 60 million people in the U.S. and over 536 million worldwide. An FDA-approved genital herpes vaccine is currently unavailable. After primary infection of the vaginal mucocutaneous tissue (VMC), the virus spreads and establishes latency in sensory neurons of regional dorsal root ganglia (DRG). The virus reactivates sporadically from latency and sheds back in the genital tract, where it can cause severe recurrent lesions. Our long-term goal is to develop a therapeutic vaccine to prevent recurrent genital herpes. Over the last 5 years, we have made significant progress in identifying candidate HSV-2 antigens and characterizing the phenotype and function of antiviral CD4+ and CD8+ T cells that associate with protection in seropositive women and in the guinea pig recurrent genital herpes model: (A) We found that two HSV-2 tegument virion proteins (VP16 and VP22) and two ribonucleotide reductase subunit proteins, (RR1 and RR2) are mainly targeted by CD4+ and CD8+ T cells from “naturally” protected asymptomatic women (those who, despite being infected, never develop recurrent genital herpes); (B) Similarly, VP16, VP22, RR1, and RR2 proteins were the main HSV-2 antigens recognized by tissue-resident CD4+ and CD8+ T cells that reside in DRG and VMC of protected asymptomatic guinea pigs; (C) Phenotypic and transcriptomic RNA- Seq profiling of DRG- and healed VMC-resident CD4+ and CD8+ T cells in protected guinea pigs show that they bear all the hallmarks of functional tissue-resident CXCR3+CD4+ and CXCR3+CD8+ T cells; (D) While therapeutic vaccination with RR2 antigen produced strong protection in HSV-2 infected guinea pigs, the VP16, VP22 and RR1 antigens provided modest protection; and (E) Treatment of HSV-2 infected guinea pigs with a neurotropic adeno-associated virus vector (AAV8) expressing the guinea pig CXCL11 chemokine (a CXCR3 ligand) boosted the number of CD4+ and CD8+ T cells specifically in infected DRG and VMC and improved protection. Based on these published and preliminary results, we hypothesize that boosting strong and long-lasting antiviral tissue- resident CD4+ and CD8+ T cell responses locally in DRG and VMC would produce a more robust/sustained protection against HSV-2 reactivation and shedding and reduce recurrent genital herpes. To test this hypothesis, we propose two Specific Aims: Aim 1. To evaluate the safety and protective efficacy, in the guinea pig genital herpes model, of an innovative prime/pull therapeutic vaccine approach that consists of: (1) Priming T cells with VP16, VP22, RR1, and RR2 antigens; and (2) “Pulling” primed T cells into infected DRG and VMC tissues by a local delivery of T-cell attracting chemokines, CXCL9, CXCL10 and/or CXCL11, using a neurotropic AAV8 vector. Aim 2. To determine whether increasing the number and function of antiviral tissue-resident CD4+ and CD8+ T cells within: (1) DRG (central neuronal immunity); and (2) VMC (peripheral epithelial immunity) correlates with protection against genital herpes. The goal of this pre-clinical study is to bring a prime/pull vaccine to clinic.

项目概要/摘要 生殖器单纯疱疹病毒 2 型 (HSV-2) 感染影响了美国超过 6000 万人，其中超过 536 人受到感染。 全球百万。目前尚无 FDA 批准的生殖器疱疹疫苗。初次感染后 阴道粘膜皮肤组织 (VMC) 中，病毒传播并在阴道粘膜皮肤组织 (VMC) 的感觉神经元中建立潜伏期 区域背根神经节（DRG）。病毒从潜伏期开始零星地重新激活，并重新回到生殖器中 道，可能导致严重的复发性病变。我们的长期目标是开发一种治疗性疫苗 预防生殖器疱疹复发。过去 5 年里，我们在识别 候选 HSV-2 抗原并表征抗病毒 CD4+ 和 CD8+ T 细胞的表型和功能 与血清阳性女性和豚鼠复发性生殖器疱疹模型的保护相关：（A）我们 发现两个 HSV-2 外皮病毒体蛋白（VP16 和 VP22）和两个核糖核苷酸还原酶亚基 蛋白质（RR1 和 RR2）主要是来自“自然”受保护的无症状患者的 CD4+ 和 CD8+ T 细胞的靶向 女性（尽管被感染，但从未出现复发性生殖器疱疹的女性）； (B) 同样，VP16、VP22、 RR1 和 RR2 蛋白是组织驻留 CD4+ 和 CD8+ T 细胞识别的主要 HSV-2 抗原 存在于受保护的无症状豚鼠的 DRG 和 VMC 中； (C) 表型和转录组 RNA- 对受保护豚鼠中 DRG 和愈合的 VMC 驻留 CD4+ 和 CD8+ T 细胞进行的序列分析表明，它们 具有功能性组织驻留 CXCR3+CD4+ 和 CXCR3+CD8+ T 细胞的所有特征； (D) 治疗期间 RR2 抗原疫苗接种对 HSV-2 感染的豚鼠、VP16、VP22 和 RR1 抗原提供了适度的保护； (E) 用神经营养剂治疗 HSV-2 感染的豚鼠 表达豚鼠 CXCL11 趋化因子（CXCR3 配体）的腺相关病毒载体 (AAV8) 增强 感染 DRG 和 VMC 中 CD4+ 和 CD8+ T 细胞的数量，并提高了保护作用。基于 根据这些已发表的初步结果，我们假设增强强大且持久的抗病毒组织- DRG 和 VMC 中局部驻留的 CD4+ 和 CD8+ T 细胞反应将产生更强大/持续的 防止 HSV-2 重新激活和脱落，并减少生殖器疱疹复发。为了检验这个假设， 我们提出两个具体目标： 目标 1. 评估豚鼠生殖器的安全性和保护功效 疱疹模型，采用创新的初免/拉动治疗性疫苗方法，包括：(1) 用 VP16、VP22、RR1 和 RR2 抗原； (2) 通过 使用神经性 AAV8 局部递送 T 细胞吸引趋化因子 CXCL9、CXCL10 和/或 CXCL11 向量。目标 2. 确定是否增加抗病毒组织驻留 CD4+ 和 CD4+ 的数量和功能 CD8+ T 细胞内： (1) DRG（中枢神经元免疫）； (2) VMC（外周上皮免疫）相关 具有预防生殖器疱疹的作用。这项临床前研究的目标是将初免/拉动疫苗推向临床。