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）

基本信息

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
项目类别：
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

项目摘要

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（由适用提供）：疟原虫（PV）是第二重要的人类疟原虫寄生虫，每年导致超过8000万例病例，包括严重的致命疾病。预防和控制受到新兴耐药性和从休眠肝寄生虫（称为催眠）的继电器的挑战。针对救济的唯一疗法，斑brimaquine会导致G6PD缺乏症患者，最普遍的人类遗传疾病的急性溶血性贫血，影响8％的疟疾 - 内态民族患者。这种治疗的障碍导致反复的PV攻击，加剧了控制问题。 Sanaria的Sporozoite（SPZ）基于PF疟疾的疫苗的高水平（100％），长期持久（至少12个月）受保护的有效性是一个重要的里程碑，这是一个重要的里程碑，这是一个重要的里程碑，并表明这种疫苗将构成一种可行的方法，用于包含和消除PF。我们认为，同样的疫苗方法可以适用于PV。在PF疫苗的开发中，受控的人类疟疾感染（CHMI）一直是进步的引擎，可以加速候选疫苗的测试。 PF CHMI最近因Sanaria的PFSPZ挑战的发展而发生了革命性的革新，无菌，纯化，冷冻保存，完全具有感染性的PFSPZ源自PF配子细胞的体外培养物，从而使志愿者在欧洲和欧洲的3个国家从未通过内部，肌内和静脉注射的志愿者成功地感染志愿者。相比之下，基于PV SPZ的产品的开发缺乏对体外培养PV寄生虫的可用技术，因此，为CHMI产生感染的蚊子，需要膜，以来自PV患者的新鲜，PV感染的血液为食。现在，我们通过使用PV Gametocyte感染的Saimiri Boliviensis非人类素数（NHP）来克服这一主要限制。实际上，我们是唯一一个由NHP感染的血液制成的小瓶中PVSPZ清单的实验室，它从2,000个蚊子中生产了多达8000万个PVSPZ。这些冷冻保存的PVSPZ在3-6天内在传统的单层格式中对肝细胞细胞系进行感染，并在12-21天的微观图案共培养的原代人肝细胞中，以及2）在VIVO中具有NHPS的感染性。现在，我们建议通过使用宽松的玻利维氏菌的特定无细菌菌落作为PV感染的血液来源，从而产生纯粹，冷冻保存，感染性PVSPZ（PVSPZ挑战）。这条新型管道将生成符合CGMP的PVSPZ批处理批次，其中包括从人类分离的各种主要和克隆PV线。 Sanaria的这一创新将提供一致的，质量控制的PVSPZ库存，以促进包括CHMI在内的PV中良好控制，可重现的体外和体内研究。这种能力技术将支持全球CHMIS抗PV药物和疫苗的开发和测试，就像PFSPZ挑战为PF CHMI所做的那样。当用抗疟疾化学预防剂（PVSPS化学预防疫苗（PVSPZ-CVAC））给药时，它还将构成一种强大的疫苗方法来预防PV疟疾的基础。