Optimizing the Generation of Monoclonal Antibodies for Prevention and Treatment of HSV Disease

优化用于预防和治疗 HSV 疾病的单克隆抗体的生成

• 批准号: 10717320
• 负责人: Betsy C. Herold
• 金额: $ 62.81万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-06-06 至 2028-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10717320
• 关键词: 2019-nCoV; Acceleration; Acute; Address; Adopted; Antibodies; Antibody-Dependent Enhancement; B-Lymphocytes; Binding; Biological Markers; Cells; Clinical; Clinical Trials; Complement; Cryoelectron Microscopy; Cytolysis; Data; Development; Disease; Dose; Ebola virus; Engineering; Epitopes; Exhibits; Female; Generations; Genital; Genitalia; Glycoproteins; Goals; Herpes Simplex Infections; Herpesvirus 1; Human; Human Herpesvirus 2; IgG Receptors; IgG1; IgG2; Immune; Immune Evasion; Immune Sera; Immune response; Immunoglobulin G; Individual; Infection; Intercellular Junctions; Interferon Type II; Knock-out; Knockout Mice; Ligands; Ligation; Light; Maps; Mediating; Modeling; Molecular; Monoclonal Antibodies; Mus; Neonatal; Pathway interactions; Phagocytosis; Play; Prevention; Production; Protein Subunits; Proteins; Recombinants; Recurrence; Role; Signal Pathway; Signal Transduction; Signaling Molecule; Simplexvirus; Skin; Structure of germinal center of lymph node; Subunit Vaccines; T-Lymphocyte; TNFRSF5 gene; Testing; Tumor Necrosis Factor-Beta; Up-Regulation; Vaccinated; Vaccination; Vaccines; Vagina; Viral; Viral Proteins; Virus; Virus Diseases; antibody-dependent cell cytotoxicity; cell type; congenic; glycoprotein D-herpes simplex virus type 2; herpesvirus entry mediator; knowledge translation; male; mouse model; neonatal mice; neonate; neutralizing antibody; novel; pathogen; preclinical study; prevent; public health priorities; receptor; response; vaccine candidate; vaccine development; vaccine-induced antibodies

There are no effective vaccines or monoclonal antibodies available for the prevention or treatment of herpes simplex viral infections. This application addresses this global public health priority. We developed a novel single-cycle vaccine strain deleted in glycoprotein D (gD), designated DgD-2. DgD-2 completely protects mice from lethal challenges with clinical isolates of both HSV-1 and HSV-2. Protection is mediated by IgG2 antibodies that have little neutralizing activity but activate Fc gamma receptors to promote antibody-dependent cellular cytotoxicity (ADCC). The central role for ADCC in mediating protection is supported by the observations that immune serum from DgD-2 vaccinated mice completely protects naïve wild-type, but not Fcg receptor IV knockout (FcgRIV-/-) mice. FcgRIV is the primary receptor responsible for mediating ADCC in mice. We isolated a highly protective monoclonal antibody (mAb), BMPC-23, from DgD-2 vaccinated mice that exhibits potent FcgRIV activating activity and mapped its epitope to domain IV of viral glycoprotein B. Notably, in contrast to the response to DgD-2, the humoral response to acute HSV infection or to vaccination with gD subunit protein vaccines in both mice and humans is neutralizing with little or no ADCC. These observations suggest that gD may interfere with the elicitation of ADCC antibodies. We hypothesized that this novel immune evasion mechanism may be mediated by interactions between gD and the co-signaling molecule switch molecule herpes virus entry mediator (HVEM), which is expressed by most immune cells. When gD binds HVEM, the interactions between HVEM and its natural ligands (LIGHT, lymphotoxin-a, CD160 and BTLA) may be inhibited. Supporting our hypothesis, IgG2 ADCC responses were reduced and protection against HSV was lost when Hvem−/− (knockout) mice were vaccinated with DgD-2. This need for HVEM to elicit IgG2 ADCC responses may be generalizable since similar results were obtained with other vaccines. Based on these data, we propose that the enhanced ADCC response to DgD-2 compared to natural HSV infection or to gD protein vaccines reflects differences in viral targets recognized, IgG subclasses generated and ability to overcome gD-HVEM mediated immune evasion. We will isolate and characterize mAbs generated in response to DgD-2, primary and recurrent HSV infection in wild-type compared to Hvem−/− mice to determine how HVEM signaling contributes to the antigenic repertoire, subclass and antibody function. We will test the mAbs that are generated in response to DgD-2 vaccination or infection individually and in combination (with BMPC-23 and other combinations) for their ability to protect and treat acute or recurrent HSV-1 and HSV-2 disease following vaginal and skin (male and female) infection as well as in models of neonatal disease. Together, these studies will yield important new fundamental and translational knowledge applicable to the development of vaccines and mAbs that mediate ADCC.

目前尚无有效的疫苗或单克隆抗体可用于预防或治疗 单纯疱疹病毒感染。该应用程序解决了这一全球公共卫生优先事项。我们开发了 一种删除了糖蛋白 D (gD) 的新型单周期疫苗株，命名为 DgD-2。 DgD-2完全 保护小鼠免受 HSV-1 和 HSV-2 临床分离株的致命挑战。保护是 由 IgG2 抗体介导，该抗体几乎没有中和活性，但可激活 Fc γ 受体 促进抗体依赖性细胞毒性（ADCC）。 ADCC 在调解中的核心作用 来自 DgD-2 接种小鼠的免疫血清完全免疫的观察结果支持了保护作用 保护幼稚野生型小鼠，但不保护 Fcg 受体 IV 敲除 (FcgRIV-/-) 小鼠。 FcgRIV 是主要受体 负责介导小鼠 ADCC。我们分离出一种高度保护性的单克隆抗体（mAb）， BMPC-23，来自 DgD-2 疫苗接种小鼠，表现出有效的 FcgRIV 激活活性，并绘制了其图谱 病毒糖蛋白 B 的结构域 IV 的表位。值得注意的是，与对 DgD-2 的反应相反，体液 小鼠和小鼠对急性 HSV 感染或接种 gD 亚单位蛋白疫苗的反应 人类在很少或根本没有 ADCC 的情况下就能发挥中和作用。这些观察结果表明 gD 可能会干扰 ADCC 抗体的引发。我们假设这种新的免疫逃避机制可能是 由 gD 和共信号分子开关分子之间的相互作用介导疱疹病毒进入 介质（HVEM），由大多数免疫细胞表达。当 gD 结合 HVEM 时，相互作用 HVEM 与其天然配体（LIGHT、淋巴毒素-a、CD160 和 BTLA）之间的相互作用可能会受到抑制。 支持我们的假设，IgG2 ADCC 反应减少，并且失去了针对 HSV 的保护作用 当 Hvem−/−（基因敲除）小鼠接种 DgD-2 疫苗时。 HVEM 需要引发 IgG2 ADCC 由于其他疫苗也获得了类似的结果，因此反应可能具有普遍性。基于这些 数据，我们建议与自然 HSV 感染或与 DgD-2 相比，ADCC 反应增强 gD 蛋白疫苗反映了识别的病毒靶标、生成的 IgG 亚类和能力的差异 克服 gD-HVEM 介导的免疫逃避。我们将分离并表征产生的单克隆抗体 与 Hvem−/− 小鼠相比，野生型小鼠对 DgD-2、原发性和复发性 HSV 感染的反应 确定 HVEM 信号传导如何影响抗原库、亚类和抗体功能。 我们将单独测试因 DgD-2 疫苗接种或感染而产生的 mAb，并 组合（与 BMPC-23 和其他组合）能够保护和治疗急性或 阴道和皮肤（男性和女性）感染后复发的 HSV-1 和 HSV-2 疾病以及 新生儿疾病模型。总之，这些研究将产生重要的新基础和 适用于开发介导 ADCC 的疫苗和单克隆抗体的转化知识。