Lung immune function in human TB infection and its perturbation by HIV-1.

NIH Pandemic-Era Grants

Pandemic Era Grants

2020

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Principal Investigator: Henry Charles Mwandumba
Organization: CORNELL UNIVERSITY
Fiscal Year: 2020
Award: $676,988
Funding agency: National Institute of Allergy and Infectious Diseases

Our recent advances in the use of fluorescent fitness reporter Mtb strains in the murine TB model has afforded
us a unique appreciation of the role of macrophage lineages in both the control and promotion of Mtb growth.
Moreover, the extension of these observations through the successful development of Dual RNA-seq protocols
for characterization of infected cells isolated directly from the murine lung has provided a further understanding
of how host nutritional immunity is central to the control of bacterial growth, at least in early infection.
 We propose building on our collaboration with Dr. Henry Mwandumba in Malawi and extending these
observations through an ex vivo Mtb challenge model with human bronchoalveolar lavage (BAL) cells to
functionally phenotype the macrophage lineages present in the human lung airways. Furthermore, we recently
demonstrated the persistence of transcriptionally-active HIV-1 genomes in the alveolar macrophages of ART-
naïve and ART-suppressed donors in Malawi and believe that we can take advantage of this unique human
subjects cohort to characterize the impairment of lung immunity known to occur in people living with HIV-1 that
renders them hypersusceptible to both TB and other lower respiratory tract infections.
 Our hypothesis is that the functional and phenotypic typing of Mtb-infected human lung macrophage
subsets from healthy and HIV-1-infected volunteers will generate testable models for immune-mediated control
of Mtb growth that will inform future vaccine development programs.
Specific Aim 1: Assessment of anti-Mtb immune function in a BAL ex vivo challenge model. This aim will
be overseen by Dr. Mwandumba in Malawi. We will use our fluorescent readouts of bacterial fitness to quantify
and optimize anti-microbial activities in human BAL cell cultures from HIV-1 uninfected and infected donors
challenged with Mtb ex vivo.
Specific Aim 2. Utilization of SILAC labeling and single cell RNA-seq to identify soluble modulators of
Mtb host macrophage function in HIV-1 negative and HIV-1 positive donors. This aim will be overseen by
Dr. Russell at Cornell University on human macrophages and BAL samples from Malawi. We will perform (i)
Proteomic analysis of released effector proteins by metabolic labeling studies (SILAC) of secreted proteins from
human BAL cells pulsed ex vivo in Malawi. (ii) We will perform single cell (scRNA-seq) RNA-seq analysis of
Mtb-challenged BAL cell populations from HIV-1 uninfected and infected donors to identify macrophage-
dependent pathways of immune control of Mtb growth and their impairment by HIV-1.
Specific Aim 3. The use of Loss of function and Gain of function approaches to assess candidate genes
and pathways in successful control of intracellular Mtb infection. We will use siRNA and synthetic mRNA
to manipulate host HMDMs and BAL macrophages to validate candidate genes/pathways involved in restriction
of bacterial growth, and how it is compromised in the HIV-1 lung environment.

Terms: <AIDS><AIDS Virus><Acquired Immune Deficiency><Acquired Immune Deficiency Syndrome><Acquired Immune Deficiency Syndrome Virus><Acquired Immuno-Deficiency Syndrome><Acquired Immunodeficiency Syndrome><Acquired Immunodeficiency Syndrome Virus><Acquired Immunologic Deficiency Syndrome><Address><Africa South of the Sahara><Alveolar Macrophages><Bacillus><Body Tissues><Bronchioalveolar Lavage><Bronchoalveolar Lavage><Bronchopulmonary Lavage><Candidate Disease Gene><Candidate Gene><Cause of Death><Cell Body><Cell Culture Techniques><Cell Lineage><Cells><Cellular Immune Function><Clinical Research><Clinical Study><Collaborations><Data><Development><Disease Progression><Environment><Equilibrium><FISH Technic><FISH Technique><FISH analysis><FISH assay><Fluorescence In Situ Hybridization><Fluorescent in Situ Hybridization><Foundations><Future><Gene Transcription><Generalized Growth><Genetic Transcription><Glycolysis><Granuloma><Granulomatous Lesion><Growth><HIV><HIV Genome><HIV-1><HIV-1 genome><HIV-I><HIV1><HIV1 genome><Human><Human Immunodeficiency Virus Type 1><Human Immunodeficiency Viruses><Human immunodeficiency virus 1><IMiD><Immune><Immune modulatory therapeutic><Immunes><Immunity><Immunize><Immunologic Model><Immunological Models><Immunomodulators><Impairment><In Vitro><Individual><Infection><Intermediary Metabolism><Knowledge><LAV-HTLV-III><Label><Link><Lower Respiratory Tract Infection><Lower respiratory infection><Lung><Lung Lavage><Lung Respiratory System><Lymphadenopathy-Associated Virus><Lymphocyte><Lymphocytic><Lymphoid Cell><M tb><M tuberculosis><M tuberculosis infection><M. tb><M. tb infection><M. tuberculosis><M. tuberculosis infection><M.tb infection><M.tuberculosis infection><MTB infection><Maintenance><Malawi><Mass Photometry/Spectrum Analysis><Mass Spectrometry><Mass Spectroscopy><Mass Spectrum><Mass Spectrum Analyses><Mass Spectrum Analysis><Mediating><Mediator><Mediator of Activation><Mediator of activation protein><Messenger RNA><Metabolic><Metabolic Processes><Metabolism><Mice><Mice Mammals><Modality><Modeling><Modern Man><Molecular><Murine><Mus><Mycobacterium tuberculosis><Mycobacterium tuberculosis (MTB) infection><Mycobacterium tuberculosis infection><Myeloid Cells><Nature><Nutritional Immunity><Nyasaland><Participant><Pathway interactions><Phenotype><Physiologic pulse><Population><Program Development><Proteins><Proteomics><Protocol><Protocols documentation><Pulmonary Macrophages><Pulse><RNA Expression><RNA Seq><RNA sequencing><RNAseq><Reporter><Role><Sampling><Short interfering RNA><Small Interfering RNA><Sub-Saharan Africa><Subsaharan Africa><TB infection><Tissue Growth><Tissues><Transcript><Transcription><Treatment Efficacy><Tuberculosis><Universities><Vaccines><Viral><Virus-HIV><active control><anti-microbial><antimicrobial><bacterial fitness><balance><balance function><bronchopulmonary lavage therapy><cell culture><co-morbid><co-morbidity><cohort><comorbidity><design><designing><develop a vaccine><development of a vaccine><developmental><disease control><disorder control><disseminated TB><disseminated tuberculosis><experiment><experimental research><experimental study><fatty acid oxidation><fetal stem cell><fitness><gain of function><global gene expression><global health><global transcription profile><human subject><immune function><immune modulating agents><immune modulating drug><immune modulating therapeutics><immune modulators><immune modulatory agents><immune modulatory drugs><immunomodulating agents><immunomodulatory agents><immunomodulatory drugs><immunomodulatory therapeutics><in vivo><infection due to Mycobacterium tuberculosis><inhibitor><inhibitor/antagonist><interstitial><intervention design><intervention efficacy><loss of function><lymph cell><mRNA><macrophage><mtb><novel><ontogeny><pathogen><pathway><permissiveness><programs><pulmonary><recruit><scRNA-seq><siRNA><single cell RNA-seq><single cell RNAseq><single-cell RNA sequencing><small molecule inhibitor><social role><therapeutic efficacy><therapeutically effective><therapy design><therapy efficacy><transcriptome><transcriptome sequencing><treatment design><tuberculosis infection><tuberculous spondyloarthropathy><vaccine development><vaccine formulation><volunteer>