Human monoclonal antibodies (huMAbs) for medically important arboviruses

在NIAID A、B和C类优先病原体虫媒病毒中，已获得批准疫苗 仅针对日本脑炎病毒、蜱传脑炎病毒和黄热病病毒（YFV）。此外，本发明还 即使是高效的YFV 17 D疫苗的使用现在也正在重新评估，因为越来越多的报告表明， 疫苗接种后的严重不良事件，特别是免疫功能低下的患者。有 目前还没有其他预防或治疗策略被批准用于虫媒病毒疾病。因此，我们建议 研究人（Hu）和人源化鼠（MuHu）虫媒病毒特异性单克隆抗体的效用， 抗体（MAb）作为抗病毒预防剂或治疗剂。初步研究，包括我们的 下面的进展报告表明，单克隆抗体可以是有效的预防和治疗剂， 脑炎性西尼罗河病毒和委内瑞拉马脑炎病毒（VEEV）在小鼠感染模型中的作用。 基于这些结果，我们假设使用Hu-或 MuHuMAb也将成功用于亲内脏的A类和C类登革热虫媒病毒（DENV）和 黄热病（YFV）我们将继续评估HuMAb对VEEV的保护和治疗能力 在我们的第一个赠款期间开发，并将使用两种方法来开发YFV和DENV 2反应 HuMAb或MuHuMAb。1)使用新的细胞融合伴侣，MFP 2或MFP 2D细胞，以制备人 使用来自感染免疫者的人外周血B细胞或人B细胞的杂交瘤 来自病毒免疫的人源化小鼠。2)基因重组保护性鼠 具有人免疫球蛋白恒定区的单克隆抗体。保护和治疗能力 将通过远交小鼠（VEEV）或自身免疫缺陷的近交系小鼠的外周病毒攻击来测试MAb。 干扰素应答（YFV和DENV 2）。具体目标如下：1.继续表征， 评估人或人源化VEEV反应性MAb和map的保护和治疗能力 人抗VEEV抗体库。2.开发YFV和DENV 2 E蛋白特异性HuMAb和MuHuMAb。 3.干扰素缺陷型YFV-17 D疫苗外周血攻毒动物模型的建立 AG 129小鼠。4.表征YFV和DENV 2反应性MuHuMAb和HuMAb的能力 以保护或治疗AG 129小鼠模型中的YFV和DENV 2感染，并使用HuMAb来定位 人抗E蛋白抗体库。该项目符合RMRCE综合研究重点 并将直接与RP 3.1和1.7以及核心C相互作用。

Of the arboviruses that are NIAID Category A, B and C Priority Pathogens, approved vaccines are available only for Japanese encephalitis virus, tick-borne encephalitis virus, and yellow fever virus (YFV). Additionally, use of even the highly effective YFV17D vaccine is now being re-evaluated due to increasing reports of severe adverse events following vaccination, particularly in immunocompromised patients. There are currently no other prophylactic or therapeutic strategies approved for arbovirus diseases. Thus, we propose to investigate the utility of human (Hu) and humanized murine (MuHu) arbovirus-specific monoclonal antibodies (MAbs) as antiviral prophylactic or therapeutic reagents. Initial studies, including those in our Progress Report below, indicate that MAbs can be effective prophylactic and therapeutic agents for the encephalitic West Nile and Venezuelan equine encephalitis viruses (VEEV) in mouse models of infection. Based on these results we hypothesize that antiviral MAb prophylaxis and/or therapy using Hu- or MuHuMAbs will also be successful for the viscerotropic, Category A and C arboviruses dengue (DENV) and yellow fever (YFV). We will continue to assess the protective and therapeutic capacities of HuMAb for VEEV developed during our first grant period and will use two approaches to develop YFV and DENV2-reactive HuMAb or MuHuMAb. 1) Use new cell fusion partners, MFP2 or MFP2D cells, to prepare human hybridomas using either human peripheral blood B-cells from infection-immune persons, or human B-cells from virus-immunized, humanized mice. 2) Genetically recombine the variable regions of protective murine MAbs with the constant regions of human immunoglobulin. Protective and therapeutic capacity of these MAbs will be tested by peripheral virus challenge of outbred mice (VEEV) or inbred mice deficient in their interferon response (YFV and DENV2). Specific aims are as follows: 1. Continue characterization and assessment of protective and therapeutic capacities of human or humanized VEEV-reactive MAbs and map the human anti-VEEV repertoire. 2. Develop HuMAbs and MuHuMAbs specific for YFV and DENV2 Eproteins. 3. Develop a new peripheral challenge animal model for YF using YFV17D vaccine in interferondeficient AG129 mice. 4. Characterize the ability of the YFV- and DENV2-reactive MuHuMAbs and HuMAbs to protect from or treat YFV and DENV2 infections in the AG129 mouse model and use the HuMAbs to map the human anti-E protein antibody repertoire. This project fits within the RMRCE Integrated Research Focus on Viral Therapeutics and will interact directly with RPs 3.1 and 1.7 and Core C.