Granzyme and Granulysin Mediated Death of Trypanosoma cruzi

颗粒酶和颗粒溶素介导的克氏锥虫死亡

基本信息

批准号：
9229525
负责人：
RICARDO TOSTES GAZZINELLI
金额：
$ 62.65万
依托单位：
BOSTON CHILDREN'S HOSPITAL
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：
2016-03-01 至 2021-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9229525
关键词：
Acute Affect Apoptosis Arrhythmia Bacteria Blood Transfusion CD8-Positive T-Lymphocytes Cardiac Cardiomyopathies Cause of Death Cell Death Cell membrane Cells Cessation of life Chagas Disease Cholesterol Chronic Cleaved cell Complex Country Cytoplasmic Granules Cytotoxic T-Lymphocytes Data Electron Transport Enzymes GZMA gene Goals Granzyme Host resistance Human Immigration Immune Immune response Infection Infiltration Inflammatory Response Interferon Type II Killer Cells Latin America Leishmania major Life Listeria monocytogenes Lymphocyte Mammalian Cell Mammals Mediating Membrane Modeling Morphology Mus Natural Immunity Natural Killer Cells Organ Transplantation Outcome Parasites Pathology Pathway interactions Patients Peptide Hydrolases Pharmaceutical Preparations Plasma Play Preventive vaccine Principal Investigator Process Proteins Public Health Role Serine Protease Severity of illness Superoxides Surrogate Markers T-Lymphocyte Testing Time Tissues Toxic Megacolon Toxoplasma gondii Transgenes Transgenic Mice Trypanosoma cruzi VDAC1 gene Vaccinated Vaccines Virulence Factors Wild Type Mouse adaptive immunity base cell killing cytotoxic disorder risk drug development fungus granulysin human disease immunological synapse in vivo insight killings macrophage neglect new therapeutic target novel pathogen perforin prevent programs public health relevance response transmission process vaccine development vaccine trial vaccine-induced immunity

项目摘要

DESCRIPTION (provided by applicant): When killer lymphocytes recognize cells infected with intracellular pathogens, they release their cytotoxic granule contents to induce apoptosis of the infected target cell. However, what happens to intracellular parasites during this process is unclear. Host cell apoptosis is triggered by cytotoxic granule proteases (granzymes, Gzm), delivered into the target cell by the cholesterol-dependent membrane perturbing protein, perforin (PFN). Cytotoxic granules of humans and some other mammals, but not rodents, contain another pore-forming protein, granulysin (GNLY), which preferentially disrupts cholesterol-poor bacterial, fungal and parasite membranes. We recently found that when human killer lymphocytes recognize bacteria-infected cells, they rapidly kill intracellular bacteria. GNLY delivers Gzms into bacteria where they proteolytically attack bacterial electron transport chain complex I to generate toxic superoxide anion and also proteolytically destroy bacterial oxidative defense enzymes. Preliminary data show that GNLY also delivers Gzms into protozoan parasites (Trypanosoma cruzi, Toxoplasma gondii, Leishmania major), which are rapidly killed, also in a superoxide-dependent manner. Destruction of intracellular parasites relies on the concerted action of all three cytotoxic effector molecules - Gzms, PFN and GNLY. Mice that express a GNLY transgene in killer lymphocytes are able to survive a T. cruzi challenge that is lethal to wild-type (WT) mice. This proposal uses T. cruzi as a model to investigate the hypothesis that GNLY, Gzm and PFN-mediated killing of intracellular protozoan parasites provides strong immune protection against intracellular parasites. We hypothesize that T. cruzi amastigotes are killed before the host cell, which limits the spread of infection. We also hypothesize that killer cells rapidly induce programs of cell death in T. cruzi that share some features with death pathways activated by Gzms in mammalian cells or bacteria. We also hypothesize that direct parasite killing helps control acute infection, enhance vaccine protection and reduce the chagasic pathology of chronic infection. This proposal joins the expertise of two principal investigators, who are leaders in studying (1) how killer cells destroy infected host cels and their pathogens and (2) the immune response to T. cruzi. The aims of the proposal are to define the features of killer cell-mediated parasite death, identify T. cruzi Gzm substrates that are important in causing death and explore how direct parasite killing contributes to immune defense by comparing infection in WT mice that do not express GNLY with GNLY-transgenic mice.

描述（由申请人证明）：当杀手淋巴细胞细胞内病原体时，它们释放了totox iC颗粒，构成靶细胞的凋亡。通过胆固醇 - 末期膜蛋白。 ES识别感染细菌的细胞，它们迅速杀死细菌。复杂的I可以生成有毒的超氧化物，以破坏细菌的氧化酶，因此将GZMS交付到寄生虫中（Cruzi，Toxoplasma gondii，Leishmania Major）杀手型淋巴细胞中的Gnly转基因能够在野生型（WT（WT））小鼠中生存，该提案使用T. Cruzi作为一个模型，以研究野生型（WT（WT））的挑战。介导的细胞内原生动物寄生虫对细胞内的寄生虫提供了强烈的免疫保护假设直接的寄生虫杀伤有助于控制慢性感染的急性渗透率，他们是研究的领导者。该提议是杀手细胞细胞寄生虫死亡蚂蚁在造成死亡中的特征，并探讨了直接寄生虫杀伤是如何通过在不表现出与gnly转基因小鼠表达的WT小鼠中构成感染来有助于免疫防御的。