Manufacture of aseptic, purified, cryopreserved Plasmodium vivax sporozoites (PvSPZ Challenge) for controlled human malaria infection (CHMI)

生产无菌、纯化、冷冻保存的间日疟原虫子孢子（PvSPZ Challenge）用于控制人类疟疾感染（CHMI）

• 批准号: 9265783
• 负责人: STEPHEN Lev HOFFMAN
• 金额: $ 30万
• 依托单位: SANARIA, INC.
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-04-21 至 2019-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9265783
• 关键词: Acute; Affect; Africa; Antimalarials; Biodistribution; Biological Assay; Bite; Blood; Cell Line; Chemoprophylaxis; Clinical trial protocol document; Communities; Country; Culicidae; Development; Disease; Dose; Drug resistance; Engineering; Equipment and supply inventories; Europe; Exhibits; Falciparum Malaria; Gabon; Geography; Germ-Free; Germany; Glucosephosphate Dehydrogenase Deficiency; Hemolytic Anemia; Hepatocyte; Hereditary Disease; Human; Human Cloning; Human Genetics; Immunity; In Vitro; Infection; International; Intramuscular Injections; Investigational New Drug Application; Kenya; Laboratories; Life; Liver; Logistics; Malaria; Manuscripts; Membrane; Mosquito Control; Netherlands; Parasitemia; Parasites; Patients; Pattern; Pharmaceutical Preparations; Phase; Plasmodium falciparum; Plasmodium falciparum vaccine; Plasmodium vivax; Prevention; Primaquine; Primates; Publishing; Quality Control; Reagent; Relapse; Reproducibility; Research; Running; Saimiri; Source; Spain; Sporozoites; Supplementation; Tanzania; Technology; Testing; Texas; Vaccines; Vial device; Vivax Malaria; Work; base; clinical toxicology; controlled release; drug relapse; feeding; germ free condition; human subject; in vivo; innovation; intradermal injection; intravenous injection; malaria infection; monolayer; mortality; nonhuman primate; novel; pathogen; permissiveness; pre-clinical; prevent; protective efficacy; public health relevance; resistant strain; success; tool; vaccine candidate; vaccine development; vaccine evaluation; volunteer

 DESCRIPTION (provided by applicant): Plasmodium vivax (Pv), the second most important human malaria parasite, causes more than 80 million cases annually including severe, fatal disease. Prevention and control are challenged by emerging drug resistance and relapses from dormant liver stage parasites called hypnozoites. The only therapy against relapse, primaquine, causes life threatening acute hemolytic anemia in patients with G6PD deficiency, the most prevalent human genetic disorder, affecting 8% of people in malaria-endemic nations. This barrier to treatment results in repeated Pv attacks, aggravating the problem of control. The demonstration of high level (100%), long-lasting (at least 12 months) protective efficacy of Sanaria's sporozoite (SPZ)-based vaccines against Pf malaria is a significant milestone, and indicates that such vaccines will constitute a viable approach to containing and eliminating Pf. We believe that the same vaccine approach could work for Pv. In the development of the Pf vaccines, controlled human malaria infection (CHMI) has been an engine of progress, accelerating the testing of vaccine candidates. Pf CHMI has recently been revolutionized by the development of Sanaria's PfSPZ Challenge, aseptic, purified, cryopreserved, fully infectious PfSPZ derived from in vitro cultures of Pf gametocytes, enabling the successful infection of volunteers by intradermal, intramuscular, and intravenous injection in 3 countries in Africa and 2 countries in Europe that had never conducted CHMI before. In contrast, development of Pv SPZ-based products has suffered from lack of available technology to culture Pv parasites in vitro, such that generating infected mosquitoes for CHMI required membrane feeding on fresh, Pv-infected blood from Pv patients. We have now overcome this major limitation by using Pv gametocyte- infected Saimiri boliviensis non-human primates (NHPs) to produce PvSPZ. In fact we are the only laboratory with an inventory of vialed PvSPZ made from NHP-infected blood, having produced as much as 80 million PvSPZ vialed in 1 day from 2,000 mosquitoes. These cryopreserved PvSPZ are 1) infectious to hepatocyte cell lines in vitro in traditional monolayer formats over 3-6 days and in micro-patterned co-cultured primary human hepatocytes over 12-21 days, and 2) infectious to NHPs in vivo. We now propose to produce aseptic, purified, cryopreserved, infectious PvSPZ (PvSPZ Challenge) by using a specific germ free colony of the permissive S. boliviensis as the source for Pv-infected blood. This novel pipeline will generate cGMP-compliant, controlled batches of PvSPZ including a wide variety of primary and clonal Pv lines isolated from humans. This innovation by Sanaria will offer a consistent, quality-controlled stock of cryopreserved PvSPZ to promote well-controlled, reproducible in vitro and in vivo studies in Pv including CHMI. This enabling technology will support the development and testing of anti-Pv drugs and vaccines in CHMIs world-wide, just as PfSPZ Challenge has done for Pf CHMIs. It will also form the basis of a powerful vaccine approach to preventing Pv malaria when administered with anti-malarial chemoprophylaxis, the PvSPZ chemoprophylaxis vaccine (PvSPZ-CVac).

 描述（由申请人提供）：间日疟原虫（Pv）是第二种最重要的人类疟疾寄生虫，每年导致超过8000万例病例，包括严重的致命疾病。预防和控制受到新出现的耐药性和休眠肝脏阶段寄生虫（称为催眠虫）复发的挑战。抗复发的唯一疗法伯氨喹在G6 PD缺乏症患者中引起危及生命的急性溶血性贫血，G6 PD缺乏症是最普遍的人类遗传性疾病，影响疟疾流行国家8%的人。这种治疗障碍导致PV反复发作，加重了控制问题。Sanaria的子孢子（SPZ）为基础的疫苗对Pf疟疾的高水平（100%），持久（至少12个月）的保护效力的证明是一个重要的里程碑，并表明这种疫苗将构成一个可行的方法来遏制和消除Pf。在Pf疫苗的开发过程中，控制人类疟疾感染（CHMI）一直是进步的引擎，加速了候选疫苗的测试。Pf CHMI最近通过Sanaria PfSPZ Challenge的开发进行了革命性的改革，PfSPZ是一种无菌、纯化、冷冻保存的完全感染性PfSPZ，来源于Pf配子母细胞的体外培养物，使之前从未进行过CHMI的3个非洲国家和2个欧洲国家的志愿者能够通过皮内、肌内和静脉注射成功感染。相比之下，基于Pv SPZ的产品的开发受到缺乏体外培养Pv寄生虫的可用技术的困扰，使得产生用于CHMI的受感染蚊子需要对来自Pv患者的新鲜的、Pv感染的血液进行膜喂养。我们现在已经通过使用Pv配子体感染的Saimiri boliviensis非人灵长类动物（NHP）来生产PvSPZ而克服了这一主要限制。事实上，我们是唯一一个拥有由NHP感染的血液制成的瓶装PvSPZ库存的实验室，在1天内从2,000只蚊子中生产了多达8000万瓶PvSPZ。这些冷冻保存的PvSPZ 1）在体外以传统单层形式感染肝细胞系3-6天，在微模式共培养的原代人肝细胞中感染12-21天，和2）在体内感染NHP。我们现在建议通过使用特定的无菌菌落的许可的S.玻利维亚人作为PV感染血液的来源。这一新的管道将产生符合cGMP的受控PvSPZ批次，包括从人类分离的各种原代和克隆Pv系。Sanaria的这一创新将提供一种稳定的、质量受控的冷冻保存PvSPZ储备，以促进包括CHMI在内的Pv的良好受控、可重现的体外和体内研究。这项技术将支持世界范围内CHMIs抗Pv药物和疫苗的开发和测试，就像PfSPZ Challenge为Pf CHMIs所做的那样。它还将形成一种强大的疫苗方法的基础，以预防Pv疟疾时，与抗疟疾化学预防，PvSPZ化学预防疫苗（PvSPZ-CVac）管理。