Mechanisms of induction of protective anti-malarial CD8+ T Cells

保护性抗疟疾 CD8 T 细胞的诱导机制

基本信息

批准号：
8631476
负责人：
MORIYA TSUJI
金额：
$ 35万
依托单位：
AARON DIAMOND AIDS RESEARCH CENTER
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-03-01 至 2018-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8631476
关键词：
Address Adenoviruses Albumins Antibodies Antigen-Presenting Cells Antigens Antimalarials Attenuated CD8B1 gene Cells Chloroquine Communicable Diseases Dendritic Cells Disease Epitopes Erythrocytes Goals Hepatocyte Human ITGAX gene Immunity Immunization In Vitro Infection Intramuscular Intravenous Kupffer Cells Lead Life Life Cycle Stages Liver Malaria Malaria Vaccines Mediating Monkeys Mus Nature Parasites Plasmodium yoelii Radiation Recombinants Rodent Role Route Societies Spleen Sporozoites Staging T cell response T-Lymphocyte Testing Transgenic Mice Transgenic Organisms Vaccination Vaccines base circumsporozoite circumsporozoite protein design improved in vivo intravenous administration macrophage mouse model promoter public health relevance subcutaneous tool vector vaccine

Address Adenoviruses Albumins Antibodies Antigen-Presenting Cells Antigens Antimalarials Attenuated CD8B1 gene Cells Chloroquine Communicable Diseases Dendritic Cells Disease Epitopes Erythrocytes Goals Hepatocyte Human ITGAX gene Immunity Immunization In Vitro Infection Intramuscular Intravenous Kupffer Cells Lead Life Life Cycle Stages Liver Malaria Malaria Vaccines Mediating Monkeys Mus Nature Parasites Plasmodium yoelii Radiation Recombinants Rodent Role Route Societies Spleen Sporozoites Staging T cell response T-Lymphocyte Testing Transgenic Mice Transgenic Organisms Vaccination Vaccines base circumsporozoite circumsporozoite protein design improved in vivo intravenous administration macrophage mouse model promoter public health relevance subcutaneous tool vector vaccine

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项目摘要

PROJECT SUMMARY/ABSTRACT Malaria remains one of the most devastating infectious diseases of the world, underscoring the need to develop effective vaccines. The current candidate malaria vaccines against the liver stages induce CD8+ T- cell-mediated protection. However, what remains unknown is the manner in which the anti-malarial CD8+ T cells are elicited in vivo. This unanswered question is particularly prominent in view of a very recent study showing that only through intravenous administration (and no other routes) do radiation-attenuated sporozoites (IrSpz) induce a potent malaria-specific CD8+ T-cell response in the livers of monkeys and of mice and provide anti-malarial protection in mice. Therefore, it appears that the nature of vaccine vectors, as well as the routes of vaccination, influences the mode of induction of "protective" anti-malarial CD8+ T cells in vivo. The overall aim of this proposal is to determine the mechanisms of in vivo induction of anti-malarial CD8+ T cells. SYVPSAEQI, derived from the P. yoelii circumsporozoite (PyCS) protein, is to date the only known CD8+ epitope that mediates "protection" against P. yoelii infection in mice and is presented by an H-2Kd molecule. Therefore, in addressing our overall goal, we have generated C57BL/6 transgenic (Tg) mice, in which Kd molecule is expressed only on dendritic cell (DC) (CD11c-Kd), macrophage (huCD68-Kd), or hepatocyte (Alb- Kd), by using CD11c promoter, huCD68 promoter, or albumin promoter, respectively. We have also generated MHC-I-Kd Tg mice that express a Kd molecule under the MHC-I promoter, in which we could induce a potent, protective anti-malarial immunity, dependent on both the PyCS protein and CD8+ T cells. These MHC-I-Kd Tg mice will be used as a positive control. In the proposed study, we will immunize the Kd Tg mice with malaria vaccines, including an adenovirus expressing the PyCS antigen, IrPySpz, or live PySpz followed by treatment with chloroquine, by different routes. We will determine the quantity, quality, and durability of PyCS antigen- specific CD8+ T-cell response induced in each group of Kd Tg mice in Aim 1. In Aim 2, we will challenge these immunized Kd Tg mice with live malaria parasites to determine the level and persistence of protective immunity induced in vivo. In Aim 3, we will determine which Kd-expressing cells induce the protective anti-malarial immunity by isolating these Kd+ cells from immunized, various Kd Tg mice, and adoptively transferring them to na¿ve MHC-I-Kd Tg mice, followed by a malaria challenge. Finally, we will isolate PyCS antigen-specific CD8+ T cells from immunized, various Kd Tg mice and adoptively transfer them to na¿ve MHC-I-Kd Tg mice, followed by a malaria challenge, to determine the protective capacity of the CD8+ T cells in Aim 4. Overall, we believe that the identification of the induction mechanisms of anti-malarial "protective" CD8+ T cells could ultimately lead to the vastly improved designs of potent T-cell-based vaccines against human malaria.

项目摘要/摘要疟疾仍然是世界上最具破坏性的传染病之一，强调了需要开发有效的疫苗。目前针对肝脏的疟疾疫苗诱导CD8+ T- 细胞介导的保护。但是，尚不清楚的是抗疟疾CD8+ T的方式细胞在体内被引起。鉴于最近的一项研究，这个未解决的问题特别突出表明只有通过静脉内给药（也没有其他路线）才能进行辐射衰减的孢子菌（IRSPZ）在猴子和小鼠的生活中诱导潜在的疟疾特异性CD8+ T细胞反应，并提供小鼠的抗疟疾保护。因此，看来疫苗向量的性质以及路线疫苗接种，影响体内“保护性”抗疟疾CD8+ T细胞的诱导方式。总体该建议的目的是确定抗疟疾CD8+ T细胞体内诱导的机制。 syvpsaeqi，源自P. yoelii圆圈（PYCS）蛋白，是迄今为止唯一已知的CD8+ 介导针对小鼠叶氏疟原虫感染的“保护”的表位，并由H-2KD分子提出。因此，在解决我们的整体目标时，我们产生了C57BL/6转基因（TG）小鼠，其中KD 分子仅在树突状细胞（DC）（CD11C-KD），巨噬细胞（HUCD68-KD）或肝细胞（Alb-- KD），分别使用CD11C启动子，HUCD68启动子或专辑启动子。我们也生成了 MHC-I-KD TG小鼠在MHC-I启动子下表达KD分子，其中我们可以诱导潜力，保护性抗疟疾免疫，取决于PYCS蛋白和CD8+ T细胞。这些MHC-I-KD TG 小鼠将被用作阳性对照。在拟议的研究中，我们将用疟疾免疫KD TG小鼠疫苗，包括表达PYCS抗原，Irpyspz或Live Pyspz的腺病毒，然后进行治疗与氯喹，通过不同的路线。我们将确定pycs抗原的数量，质量和耐用性在AIM 1中，每组KD TG小鼠诱导的特定CD8+ T细胞响应。在AIM 2中，我们将挑战这些具有活疟疾寄生虫的免疫化KD TG小鼠，以确定受保护的免疫组织化学的水平和持久性诱导体内。在AIM 3中，我们将确定哪些表达KD的细胞会诱导受保护的抗疟疾通过从免疫，各种KD TG小鼠中分离这些KD+细胞，并将其自适应地转移到 NA¿VEMHC-I-KD TG小鼠，随后是疟疾挑战。最后，我们将隔离PYCS抗原特异性CD8+ 来自免疫，各种KD TG小鼠的T细胞，并将其自适应地转移到NA¿VEMHC-I-KD TG小鼠中通过疟疾挑战，确定AIM 4中CD8+ T细胞的保护能力。总体而言，我们相信鉴定抗疟疾“保护性” CD8+ T细胞的诱导机制最终可能导致对人类疟疾的潜在基于T细胞的疫苗的设计大大改进。