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; 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）： 总结项目大多数暴露于结核分枝杆菌的人会被感染并发展为潜在的MTB感染。但是，在乌干达城市的结核病联系研究中，在随访的两年中，大约9％的亲密接触没有发展LTBI。在非洲，一些矿工在世界上最高的MTB感染压力下，没有超过20年的LTBI目的地就业。这些人没有LTBI目的地强烈的MTB暴露的证据似乎是LTBI撤退者（RSTRS）。 MTB感染后的单核细胞揭示了将RSTR与HHC与LTBI区分开的转录特征。这些数据支持以下假设：宿主因素可以区分RSTR，并且它们保护了在巨噬细胞中表达的先天免疫调查剂。 HIV对MTB的免疫调查失调，因此，感染HIV的人更多地依赖于先天免疫来控制MTB，而不是HIV人。我们假设在HIV+ RSTR中更容易鉴定出对LTBI抗性的先天机制。目的是：1。识别，特征和招募艾滋病毒+个体的人群，他们抵抗乌干达结核病家庭接触和南非矿工之间的潜在MTB感染。方法：以前从乌干达的结核病HHC中识别出艾滋病毒+ RSTR的后续行动，并招募了新的RSTR，而我S. Africa招募了新的HIV+ RSTR。 HIV+ LTBIS，HIV-RSTR和HIV-LTBI将作为对照。假设：在具有不同MTB暴露的人群中可以鉴定HIV+和HIV-RSTR，但将共享宿主抗性机制，从而使稳健的同伙用于先天免疫功能研究。 AIM 2。确定与HIV+患者对潜在MTB感染的抗性有关的巨噬细胞转录特征。方法：比较HIV+ RSTR和LTBI之间巨噬细胞中对MTB的早期转录反应。将这些与HIV-RSTR和HIV-LTBIS的巨噬细胞的反应进行比较，并将研究扩展到肺泡巨噬细胞。假设：巨噬细胞中的生物学途径调节对MTB感染的抗性，这些途径在HIV+患者中更容易鉴定出来。 AIM 3。确定对潜在MTB感染抗性的细胞机制。方法：通过CRISPR/CAS9技术和小分子抑制剂进行细胞敲除，以研究AIM 2中鉴定的关键基因和途径如何使巨噬细胞抵抗或清除MTB； b。 MTB杀死的全血分析，以区分HIV+ RSTR和LTBIS； c。流式细胞仪检查是否在HIV+ RSTR中富含先天的非常规的T细胞。假设： HIV+ RSTR使用巨噬细胞特异性途径来控制MTB； b。 WBA可以检测RSTR和LTBI之间MTB生长的控制差异。 c。 HIV+ RSTRS使用非常规的先天T细胞来控制MTB。在这个项目中，我们将两种经验结合了TB临床研究网站（乌干达的Mulago医院和非洲的Aurum Institute）与流行病学家，系统生物学家，遗传学家，免疫学家，微生物学家，微生物学家和临床医生在乌干达大学的Makerere University，S. novara和Case Waster anton和案例。