Resistance to MTB infection in HIV infected individuals in Uganda and S. Africa

乌干达和南非 HIV 感染者对 MTB 感染的抵抗力

• 批准号: 9495556
• 负责人: W. Henry Boom
• 金额: $ 242.22万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-06-01 至 2021-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9495556
• 关键词: Address; Adult; Africa; Alveolar Macrophages; Biological; Biological Assay; Biological Markers; Blood; CD4 Positive T Lymphocytes; CRISPR/Cas technology; Cell Count; Cells; Clinical; Communicable Diseases; Computing Methodologies; Data; Disease; Employment; Epidemiologist; Exposure to; Flow Cytometry; Fred Hutchinson Cancer Research Center; Gene Expression Profile; Genes; Genetic Transcription; Growth; HIV; HIV Infections; HIV-1; Hospitals; Host resistance; Household; Human; Immune response; Immunologist; Individual; Innate Immune Response; Institutes; Integration Host Factors; Interferon-alpha; Knock-out; Lung; Measures; Mining; Mononuclear; Mycobacterium tuberculosis; Natural Immunity; Pathogenesis; Pathway interactions; Persons; Phagocytes; Phenotype; Physicians; Plasma; Population; Recording of previous events; Resistance; South Africa; System; Systems Biology; T-Lymphocyte; Tuberculin Test; Tuberculosis; Tuberculosis Vaccines; Uganda; Universities; Viral Load result; Washington; Whole Blood; aerosolized; antiretroviral therapy; clinical phenotype; clinical research site; cohort; experience; follow-up; genome-wide linkage; innate immune function; insight; macrophage; monocyte; multidisciplinary; novel strategies; pathogen; peripheral blood; pressure; public health relevance; recruit; resistance mechanism; response; small molecule inhibitor; transcriptome sequencing; uptake

 DESCRIPTION (provided by applicant): PROJECT SUMMARY Most people heavily exposed to Mycobacterium tuberculosis become infected and develop latent Mtb infection. However, in a TB contact study in urban Uganda, approximately 9% of close adult contacts did not develop LTBI during two years of follow-up. In S. Africa, some miners do not develop LTBI despite more than 20 years of employment under the highest Mtb infection pressure in the world. These persons without evidence of LTBI despite intense Mtb exposure appear to be LTBI resisters (RSTRs). Monocytes after Mtb infection revealed transcriptional signatures that distinguished RSTRs from HHC with LTBI. These data support the hypothesis that host factors can distinguish RSTRs and that they have protective innate immune responses expressed in macrophages. HIV dysregulates immune responses to Mtb, thus HIV infected persons depend more on innate immunity to control Mtb than HIV- persons. Thus we hypothesize that innate mechanisms of resistance to LTBI are more readily identified in HIV+ RSTRs. The aims are: 1. Identify, characterize and recruit cohorts of HIV+ individuals who resist latent Mtb infection among TB household contacts in Uganda and among miners in South Africa. Approach: Follow-up previously identified HIV+ RSTRs and recruit new RSTRs from among TB HHC in Uganda, and i S. Africa recruit a new cohort of HIV+ RSTRs. HIV+ LTBIs, and HIV- RSTRs and HIV- LTBIs will serve as controls. Hypothesis: HIV+ and HIV- RSTRs can be identified in populations with different Mtb exposures but host resistance mechanisms will be shared and thus enable assembly of robust cohorts for innate immune function studies. Aim 2. Determine macrophage transcriptional signatures associated with resistance to latent Mtb infection in HIV+ persons. Approach: Compare early transcriptional responses to Mtb in macrophages between HIV+ RSTRs and LTBIs. Compare these to the responses of macrophages from HIV- RSTRs and HIV- LTBIs, and extend studies to alveolar macrophages. Hypothesis: Biological pathways in macrophages regulate resistance to Mtb infection and these pathways are more readily identified in HIV+ persons. Aim 3. Determine cellular mechanisms of resistance to latent Mtb infection. Approach: a. Cellular knockouts through CRISPR/Cas9 technology and small molecule inhibitors to investigate how key genes and pathways identified in Aim 2 allow macrophages to resist or clear Mtb; b. Whole blood assays of Mtb killing to distinguish HIV+ RSTRs from LTBIs; c. Flow cytometry to examine whether innate unconventional T-cells are enriched in HIV+ RSTRs. Hypothesis: a. HIV+ RSTR use macrophage-specific pathways to control Mtb; b. A WBA can detect differences in control of Mtb growth between RSTRs and LTBIs; c. HIV+ RSTRs use unconventional innate T cells to control Mtb. For this project, we have assembled two experienced TB clinical research sites (Mulago Hospital in Uganda & the Aurum Institute in S. Africa) with epidemiologists, systems biologists, geneticists, immunologists, microbiologists, and clinicians at Makerere University in Uganda, Aurum Institute in S. Africa, University of Washington and Case Western Reserve University.

 描述（由申请人提供）： 项目摘要 大多数严重接触结核分枝杆菌的人都会受到感染并出现潜伏性结核分枝杆菌感染，但在乌干达城市的一项结核病接触研究中，大约 9% 的密切成人接触者在两年的随访期间并未出现 LTBI。在非洲，尽管在世界上最高的结核菌感染压力下工作了 20 多年，但一些矿工并未患上潜伏性结核病。尽管这些人大量接触结核菌，但没有感染潜伏性结核病的证据。 Mtb 感染后的 Mon 揭示了将 RSTR 与 HHC 与 LTBI 区分开来的假设，即宿主因素可以区分 RSTR，并且它们具有在巨噬细胞中表达的保护性先天免疫反应。结核分枝杆菌，因此艾滋病毒感染者比艾滋病毒感染者更依赖先天免疫来控制结核分枝杆菌，因此我们面临着更容易识别结核分枝杆菌的先天机制。 HIV+ RSTR 的目标是： 1. 在乌干达的结核病家庭接触者和南非矿工中识别、描述和招募能够抵抗潜伏 Mtb 感染的 HIV+ 个体。乌干达和南非的结核病 HHC 中招募了一批新的 HIV+ RSTR，HIV+ LTBI，HIV-RSTR 和 HIV-LTBI 将作为他们的成员。假设：可以在具有不同 Mtb 暴露的人群中鉴定出 HIV+ 和 HIV- RSTR，但宿主抵抗机制将被共享，从而能够组装强大的群体以进行先天免疫功能研究。 目标 2. 确定与潜在抵抗相关的巨噬细胞转录特征。方法：比较 HIV+ RSTR 和 LTBI 之间巨噬细胞对 Mtb 的早期转录反应。假设：巨噬细胞中的生物途径调节对 Mtb 感染的抵抗力，并且这些途径在 HIV+ 人群中更容易识别。目标 3. 确定对潜伏 Mtb 抵抗力的细胞机制。方法：a. 通过 CRISPR/Cas9 技术和小分子抑制剂进行细胞敲除，以研究 Aim 2 中确定的关键基因和途径如何实现。巨噬细胞抵抗或清除 Mtb；b. 杀灭 Mtb，以区分 HIV+ RSTR 和 LTBI；c. 流式细胞术检查先天非常规 T 细胞是否富含 HIV+ RSTR。控制 Mtb；b. WBA 可以检测 RSTR 和 LTBI 之间 Mtb 生长控制的差异； c. HIV+ RSTR 使用非常规的先天 T 细胞来控制结核分枝杆菌。在这个项目中，我们聚集了两个经验丰富的结核病临床研究中心（乌干达的穆拉戈医院和南非的 Aurum 研究所），其中包括流行病学家、系统生物学家、遗传学家、免疫学家、乌干达麦克雷雷大学、南非奥鲁姆研究所、华盛顿大学和凯斯西储大学的微生物学家和新人。