Antibody therapeutics against biodefense-related diseases

针对生物防御相关疾病的抗体疗法

• 批准号: 8349357
• 负责人: Dimiter S Dimitrov
• 金额: $ 24.13万
• 依托单位: DIVISION OF BASIC SCIENCES - NCI
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8349357
• 关键词: Active Immunization; Acute; Animal Model; Animal Testing; Animals; Antibiotic Resistance; Antibiotics; Antibodies; Antigens; Antiviral Agents; Bacteremia; Binding; Calcium; Capsid; Categories; Clinical; Communicable Diseases; Crimean-Congo Hemorrhagic Fever Virus; Dengue; Dengue Hemorrhagic Fever; Dengue Virus; Diagnosis; Diagnostic; Disease; Ebola virus; Epitopes; Exhibits; FDA approved; Frankfurt-Marburg Syndrome Virus; Glycoproteins; Goals; Hendra Virus; Henipavirus; Human; Incidence; Infection; Libraries; Microbe; Modeling; Monoclonal Antibodies; Mus; National Institute of Allergy and Infectious Disease; Nipah Virus; Palivizumab; Passive Immunization; Patients; Peptides; Phage Display; Plague; Prevention; Prophylactic treatment; Proteins; Reagent; Recombinants; Recording of previous events; Reporting; Research; Surface; Testing; Therapeutic; Therapeutic antibodies; Time; Toxin; Vaccines; Viral Hemorrhagic Fevers; Virulent; Virus; Yersinia pestis; animal model development; base; biodefense; biothreat; human monoclonal antibodies; in vitro testing; interest; killings; mortality; mouse model; neutralizing monoclonal antibodies; novel; pathogen; prophylactic; research and development; research study; resistant strain; response; therapeutic vaccine; tool development

We have continued to identify and characterize neutralizing mAbs to the following biodefense-related microbes which are on the NIAID list of category A or C pathogens: Nipah and Hendra viruses (Henipaviruses) (causing acute infections with high, up to 70%, mortality, category C), Ebola and Marburg viruses (causing hemorrhagic fever with high, up to 90%, mortality, category A), Crimean-Congo virus (CCHFV) (causing hemorrhagic fever with relatively high, up to 50%, mortality, category C), and dengue virus (which causes dengue hemorrhagic fever with relatively low mortality, category A). 1) Previously, we reported the isolation of henipavirus-neutralizing recombinant hmAbs including one, m102.4, which exhibited exceptionally potent and cross-reactive inhibitory activity against both HeV and NiV. HeV is categorized as a biosafety level 4 agent, which has made development of animal models and testing of potential therapeutics and vaccines challenging. The results from two animal studies and three humans administered with this mAb confirm our proposition that m102.4 has potential as a therapeutic for treatment of diseases caused by henipaviruses, and could save human lives now. It could be also used for prophylaxis, diagnosis and as a research reagent. Animal studies with this antibody continue and more are planned with GMP produced m102.4. 2) We have previously identified several novel potent hmAbs against Yersinia pestis which are the first human antibodies against this category A agent. Yersinia pestis is the etiologic agent of plague that has killed more than 200 million people throughout the recorded history of mankind. Antibiotics may provide little immediate relief to patients who have a high bacteremia or to patients infected with an antibiotic resistant strain of plague. Two virulent factors of Y. pestis are the capsid F1 protein and the low-calcium response (Lcr) V-protein or V-antigen that have been proven to be the targets for both active and passive immunization. There are mouse monoclonal antibodies (mAbs) against the F1- and V-antigens that can passively protect mice in a murine model of plague; however, there are no anti-Yersinia pestis monoclonal antibodies available for prophylactic or therapeutic treatment in humans. We identified one anti-F1-specific human mAb (m252) and two anti-V-specific human mAb (m253, m254) by panning a nave phage-displayed Fab library against the F1- and V-antigens. The Fabs were converted to IgG1s and their binding and protective activities were evaluated. M252 bound weakly to peptides located at the F1 N-terminus where a protective mouse anti-F1 mAb also binds. M253 bound strongly to a V-antigen peptide indicating a linear epitope; m254 did not bind to any peptide from a panel of 53 peptides suggesting that its epitope may be conformational. M252 showed better protection than m253 and m254 against a Y, pestis challenge in a plague mouse model. A synergistic effect was observed when the three antibodies were combined. Incomplete to complete protection was achieved when m252 was given at different times post-challenge. These antibodies can be further studied to determine their potential as therapeutics or prophylactics in Y. pestis infection in humans. 3) We have also performed experiments to identify neutralizing hmAbs against dengue virus and Ebola virus. Several mAbs against dengue virus were identified and are being characterized. The mAbs against dengue could be used as candidate therapeutics and diagnostics and as tools for development of vaccine immunogens.

我们继续鉴定和鉴定针对以下生物防御相关微生物的中和单克隆抗体，这些微生物属于 NIAID A 类或 C 类病原体清单：尼帕病毒和亨德拉病毒（亨尼帕病毒）（引起死亡率高达 70% 的急性感染， C 类）、埃博拉病毒和马尔堡病毒（引起出血热，死亡率高达 90%，A 类）、克里米亚-刚果病毒(CCHFV)（引起出血热，死亡率相对较高，高达 50%，C 类）和登革热病毒（引起登革出血热，死亡率相对较低，A 类）。 1) 之前，我们报道了亨尼帕病毒中和重组 hmAb 的分离，其中包括一种 m102.4，它对 HeV 和 NiV 表现出异常有效的交叉反应抑制活性。 HeV 被归类为生物安全 4 级制剂，这使得动物模型的开发以及潜在疗法和疫苗的测试变得具有挑战性。两项动物研究和三名人类使用该单克隆抗体的结果证实了我们的观点，即 m102.4 有潜力作为治疗亨尼帕病毒引起的疾病的药物，并且现在可以挽救人类的生命。它还可用于预防、诊断和作为研究试剂。使用该抗体的动物研究仍在继续，并计划使用 GMP 生产的 m102.4 进行更多动物研究。 2) 我们之前已经鉴定出几种针对鼠疫耶尔森氏菌的新型强效 hmAb，这是针对这种 A 类病原体的第一个人类抗体。鼠疫耶尔森氏菌是鼠疫的病原体，在人类有记载的历史中，已导致超过 2 亿人死亡。对于菌血症高的患者或感染抗生素耐药鼠疫株的患者，抗生素可能无法立即缓解症状。鼠疫耶尔森氏菌的两个毒力因子是衣壳 F1 蛋白和低钙反应 (Lcr) V 蛋白或 V 抗原，已被证明是主动和被动免疫的目标。针对 F1 和 V 抗原的小鼠单克隆抗体 (mAb) 可以被动保护鼠疫小鼠模型中的小鼠；然而，尚无可用于人类预防性或治疗性治疗的抗鼠疫耶尔森氏菌单克隆抗体。我们通过针对 F1 和 V 抗原淘选天然噬菌体展示的 Fab 文库，鉴定了一种抗 F1 特异性人 mAb (m252) 和两种抗 V 特异性人 mAb (m253、m254)。 Fab 被转化为 IgG1，并评估它们的结合和保护活性。 M252 与位于 F1 N 末端的肽微弱结合，保护性小鼠抗 F1 mAb 也与该肽结合。 M253 与 V 抗原肽强烈结合，表明其为线性表位； m254 不与 53 个肽组中的任何肽结合，表明其表位可能是构象的。在鼠疫小鼠模型中，M252 对鼠疫耶尔森氏菌攻击表现出比 m253 和 m254 更好的保护作用。当三种抗体组合时观察到协同效应。当在攻击后的不同时间给予 m252 时，实现了不完全到完全的保护。可以进一步研究这些抗体，以确定它们作为人类鼠疫耶尔森氏菌感染的治疗剂或预防剂的潜力。 3) 我们还进行了实验来鉴定针对登革热病毒和埃博拉病毒的中和 hmAb。已鉴定出几种针对登革热病毒的单克隆抗体，并正在对其进行表征。针对登革热的单克隆抗体可用作候选治疗和诊断方法以及开发疫苗免疫原的工具。