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。该申请应解决此全球公共卫生优先事项。我们开发了 在糖蛋白D（GD）中删除的一种新型的单周期疫苗菌株，指定为DGD-2。 DGD-2完全 保护小鼠免受HSV-1和HSV-2的临床分离株的致命挑战。保护是 由IgG2抗体介导的，这些抗体几乎没有中和活性，但会激活FC伽马受体 促进抗体依赖性细胞毒性（ADCC）。 ADCC在调解中的核心作用 保护完全来自DGD-2接种小鼠的免疫血清的观察结果支持了保护 保护幼稚的野生型，但不能保护FCG受体IV敲除（FCGRIV - / - ）小鼠。 FCGRIV是主要受体 负责介导小鼠ADCC。我们分离了高度保护的单克隆抗体（MAB）， BMPC-23，来自DGD-2疫苗接种小鼠，该小鼠具有潜在的FCGRIV激活活性并映射 病毒糖蛋白B的域IV的表位B.特别是与对DGD-2的反应相比， 对急性HSV感染的反应或在小鼠和 人类很少或没有ADCC中和。这些观察结果表明，GD可能会干扰 促进ADCC抗体。我们假设这种新型免疫进化机制可能是 由GD与共同信号分子开关分子疱疹进入的相互作用介导 介体（HVEM），大多数免疫细胞表达。当GD绑定HVEM时，相互作用 在HVEM及其天然配体（光，淋巴毒素-A，CD160和BTLA）之间。 支持我们的假设，IgG2 ADCC响应减少了，对HSV的保护丢失了 当HVEM - / - （敲除）小鼠接种DGD-2时。 HVEM引起IgG2 ADCC的需求 由于用其他疫苗获得了相似的结果，因此可以推广反应。基于这些 数据，我们提出，与天然HSV感染相比，ADCC对DGD-2的反应增强了或 GD蛋白疫苗反映了识别的病毒靶标的差异，产生的IgG亚类和能力 克服GD-HVEM介导的免疫进化。我们将隔离并表征生成的mAb 与HVEM - / - 小鼠相比 确定HVEM信号如何有助于抗原库，亚类和抗体功能。 我们将测试针对DGD-2疫苗接种或感染而单独以及 组合（与BMPC-23和其他组合）保护和治疗急性或 阴道和皮肤（雄性和女性）感染后，复发性HSV-1和HSV-2疾病以及 新生儿疾病的模型。这些研究将共同​​产生重要的新基本和 转化知识适用于介导ADCC的疫苗和mAB的开发。