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.

没有有效的疫苗或单克隆抗体可用于预防或治疗 单纯疱疹病毒感染本申请涉及这一全球公共卫生优先事项。我们开发 一种新型糖蛋白D（gD）缺失的单循环疫苗株，命名为DgD-2。DgD-2完全 保护小鼠免受HSV-1和HSV-2临床分离株的致命攻击。保护是 由IgG 2抗体介导，其几乎没有中和活性，但激活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免疫小鼠的免疫血清完全中和了DgD-2的免疫保护作用。 保护幼稚野生型，但不保护Fcg受体IV敲除（FcgRIV-/-）小鼠。FcgRIV是主要受体 在小鼠中负责介导ADCC。我们分离了一种高度保护性的单克隆抗体（mAb）， BMPC-23，来自DgD-2接种的小鼠，其表现出有效的FcgRIV活化活性，并将其定位于 病毒糖蛋白B的结构域IV的表位。值得注意的是，与对DgD-2的应答相反，体液免疫应答与DgD-2的应答相反。 在小鼠中对急性HSV感染或对gD亚单位蛋白疫苗接种的应答， 人类是中和的，几乎没有ADCC。这些观察结果表明，gD可能会干扰 引发ADCC抗体。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的疱疹病毒进入 介体（HVEM），其由大多数免疫细胞表达。当gD结合HVEM时， HVEM与其天然配体（LIGHT、光敏素-a、CD 160和BTLA）之间的相互作用可以被抑制。 支持我们的假设，IgG 2 ADCC反应降低，对HSV的保护作用丧失 当Hvem−/−（敲除）小鼠接种DgD-2时。HVEM需要引发IgG 2 ADCC 由于用其他疫苗获得了类似的结果，因此应答可以是可推广的。基于这些 数据，我们提出，增强ADCC反应DgD-2相比，自然HSV感染或 gD蛋白疫苗反映了识别的病毒靶标、产生的IgG亚类和能力的差异。 以克服gD-HVEM介导的免疫逃避。我们将分离和表征在 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信号传导如何有助于抗原库、亚类和抗体功能。 我们将单独测试响应DgD-2疫苗接种或感染产生的mAb， 与BMPC-23和其它组合物联合使用，以保护和治疗急性或 阴道和皮肤（男性和女性）感染后的复发性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的基础和 适用于开发介导ADCC的疫苗和mAb的翻译知识。