Translation Research in Plasmodium vivax

本提案中描述的研究目的是鉴定和表征针对来自寄生虫红细胞前期和蚊子阶段的已定义和新发现的间日疟原虫抗原的保护性免疫反应。然后配制诱导这些免疫应答并在Aotus猴模型中提供保护的疫苗。(A)将在终生暴露于间日疟原虫疟疾的流行区居民以及用辐照间日疟原虫子孢子免疫的志愿者中确定被认为与保护有关的免疫应答，并显示出对子孢子攻击的保护。(b)将通过评估Aotus猴和人类志愿者的寄生虫血症在体内进行保护性生物活性检测，并通过评估血清抑制肝细胞子孢子侵入、肝细胞培养物中子孢子发育和血液阶段培养物中血液阶段发育的能力在体外进行保护性生物活性检测，并通过评估饲喂含配子体的血液阶段培养物的白曼按蚊的卵囊发育进行保护性生物活性检测。(c)来自受保护和未受保护个体的血清将通过子孢子、肝脏和血液阶段IFAT和通过利用选择的全长重组蛋白、长肽或组分表位的ELISA来表征。我们将首先关注可用的试剂：PvSP和PvSSP 2/TRAP（子孢子和肝阶段抗原），PvMSP-1和PvDBP（血液阶段抗原）和Pvs 25/28（蚊子肠道中表达的抗原）。其他抗原将在获得后进行研究。淋巴细胞将通过ELISPOT响应于源自PvCSP和PvSSP 2（以及源自与各种佐剂、DNA质粒和重组痘病毒和猴模型一起配制的蛋白质或肽）的表位的CTL和干扰素γ产生来表征。我们将利用Aotus的血液阶段攻毒模型和Aotus中新开发的子孢子攻毒模型。该研究项目将综合利用过去十年在哥伦比亚西部精心开发的实验室、灵长类动物设施、昆虫实验室和实地考察点，以回答关于这种重要但相对被忽视的疟原虫的关键生物学问题。

The purpose of the research described in this proposal is to identify and characterize protective immune responses against defined and newly discovered Plasmodium vivax antigens from the pre-erythrocytic and mosquito stages of the parasite., and then to formulate vaccines that induce these immune responses and provide protection in the Aotus monkey model. (A) Immune responses thought to be involved in protection will be identified in residents of endemic areas with life-long exposure to P. vivax malaria and also in volunteers immunized with irradiated P. vivax sporozoites and shown to be protected against sporozoite challenge. (b) Testing for protective biological activity will be done in vivo by assessing parasitemia in Aotus monkeys and in human volunteers, and in vitro by assessing the ability of sera to inhibit sporozoite invasion of hepatocytes, development of sporozoites in hepatocyte culture, and development of blood stages in blood stage culture, and by assessing oocyst development in Anopheles albimanus fed on blood stage cultures containing gametocytes. (c) Sera from protected and non protected individuals will be characterized by sporozoite, liver and blood stage IFAT and by ELISA utilizing selected full length recombinant proteins, long peptides or components epitopes. We will focus initially on reagents that are available: PvSP and PvSSP2/TRAP (sporozoite and liver stage antigens), PvMSP-1 and PvDBP (blood stage antigens) and Pvs25/28 (antigens expressed in the mosquito gut). Other antigens will be studied as they become available. Lymphocytes will be characterized by CTL and interferon gamma production by ELISPOT in response to epitopes derived from the PvCSP and PvSSP2 (and from proteins or peptides formulated with various adjuvants, DNA plasmids, and recombinant poxviruses, and monkey models. We will utilize the blood stage challenge model in Aotus and a newly developed sporozoite challenge model in Aotus. This research project will integrate the use of laboratories, primate facilities, insectories and field sites painstakingly developed over the last decade in western Columbia in order to answer these critical biological questions about this important but relatively neglected malaria parasite.