H Mwandumba, CoM, Characterisation of the breakdown in immune competence of the lung that favours development of tuberculosis in HIV-infected adults
Lead Research Organisation:
Liverpool School of Tropical Medicine
Department Name: Clinical Sciences
Abstract
Tuberculosis (TB) remains a major health problem in sub-Saharan Africa where the morbidity and mortality due to this infection are high, predominantly as a consequence of human immunodeficiency virus (HIV) infection. While it is unequivocal that infection with HIV results in loss of immune control of Mycobacterium tuberculosis (Mtb) by the host, it is unclear if the increased risk of TB disease is due solely to depletion of CD4+ T-cells (a type of immune cells), or due to the loss of other immune functions. For instance, the risk of developing active TB starts to increase soon after HIV infection, even before significant depletion of CD4+ T-cells in peripheral blood. Furthermore, antiretroviral therapy (ART)-mediated immune recovery does not eliminate the increased risk of TB in HIV-infected individuals, indicating that reconstitution of the CD4+ T-cell subset alone is not sufficient to recover full protection against active TB. Accumulating evidence also indicates that in regions of high Mtb transmission, many TB cases in HIV-infected individuals result from recent Mtb infections and not reactivation of pre-existing infection. Since the majority of Mtb infections are acquired through the respiratory route, this suggests strongly that the lung environment in HIV-infected individuals is altered to permit the establishment and progression of new Mtb infections.
In view of existing limitations in current understanding of the pulmonary protective immune response against Mtb infections in humans, this project will utilize human lung immune cells and ex vivo experimental infections with characterized, reporter Mtb strains to probe the mechanistic basis underlying the successful control of Mtb in the lung. It will also explore the impact of HIV co-infection and ART on immune competence of the lung in general, and the Mtb-limiting immune response
in particular. The project will capitalize on the long-standing links between the Liverpool School of Tropical Medicine, Cornell University and the Malawi-Liverpool-Wellcome Trust Clinical Research Programme (MLW)'s strongly integrated clinical research base, high quality laboratories and excellent training infrastructure. It will strengthen existing collaborations and forge new ones to promote the development and transfer of cutting-edge technology to Malawi. By exploiting the HIV-mediated breakdown in immune control of Mtb, the project is bold and highly innovative in its approach to gain a comprehensive understanding of the optimal pulmonary immune environment for successful control of this pathogen. It will charter new areas of investigation and has tremendous potential to contribute to the field important new insights that will impact the design and implementation of public health interventions for control of TB in both HIV-uninfected and HIV-infected individuals.
In view of existing limitations in current understanding of the pulmonary protective immune response against Mtb infections in humans, this project will utilize human lung immune cells and ex vivo experimental infections with characterized, reporter Mtb strains to probe the mechanistic basis underlying the successful control of Mtb in the lung. It will also explore the impact of HIV co-infection and ART on immune competence of the lung in general, and the Mtb-limiting immune response
in particular. The project will capitalize on the long-standing links between the Liverpool School of Tropical Medicine, Cornell University and the Malawi-Liverpool-Wellcome Trust Clinical Research Programme (MLW)'s strongly integrated clinical research base, high quality laboratories and excellent training infrastructure. It will strengthen existing collaborations and forge new ones to promote the development and transfer of cutting-edge technology to Malawi. By exploiting the HIV-mediated breakdown in immune control of Mtb, the project is bold and highly innovative in its approach to gain a comprehensive understanding of the optimal pulmonary immune environment for successful control of this pathogen. It will charter new areas of investigation and has tremendous potential to contribute to the field important new insights that will impact the design and implementation of public health interventions for control of TB in both HIV-uninfected and HIV-infected individuals.
Technical Summary
Co-infection with Mycobacterium tuberculosis (Mtb) and human immunodeficiency virus (HIV) constitutes a huge burden on national healthcare services in sub-Saharan Africa where up to 70% of tuberculosis (TB) patients are HIV-infected. TB is now the leading cause of death of adults living with HIV in this region. This underscores the need for a better understanding of the mechanisms underlying host resistance and susceptibility to TB, which is central to the development of new approaches to prevent and/or treat HIV-associated TB.
The risk of developing TB starts to increase early following HIV infection, even before significant depletion of CD4 T lymphocytes occurs in peripheral blood and persists during effective antiretroviral therapy (ART), indicating that the CD4 T lymphocyte count is not the only determinant of increased TB risk, and ART-mediated quantitative reconstitution of this cell subset alone is not sufficient to recover full protection against active TB.
We propose conducting ex vivo infections of human alveolar macrophages (AMs) with fluorescent reporter Mtb strains to probe the mechanistic basis underlying the successful control of Mtb infection in the lung. The use of reporter bacterial strains is significant because they are immunologically unbiased readouts of host cell function. We will explore the impact of HIV co-infection and ART on the AM anti-mycobacterial response. By exploiting the HIV-mediated breakdown in immune control of Mtb infection in the lung, the project is bold and highly innovative in its approach to gain a comprehensive understanding of the optimal pulmonary immune environment for successful control of Mtb infection. It has tremendous potential to generate new insights that will impact the design and implementation of public health interventions in the control of TB in both HIV-uninfected and HIV-infected individuals.
The risk of developing TB starts to increase early following HIV infection, even before significant depletion of CD4 T lymphocytes occurs in peripheral blood and persists during effective antiretroviral therapy (ART), indicating that the CD4 T lymphocyte count is not the only determinant of increased TB risk, and ART-mediated quantitative reconstitution of this cell subset alone is not sufficient to recover full protection against active TB.
We propose conducting ex vivo infections of human alveolar macrophages (AMs) with fluorescent reporter Mtb strains to probe the mechanistic basis underlying the successful control of Mtb infection in the lung. The use of reporter bacterial strains is significant because they are immunologically unbiased readouts of host cell function. We will explore the impact of HIV co-infection and ART on the AM anti-mycobacterial response. By exploiting the HIV-mediated breakdown in immune control of Mtb infection in the lung, the project is bold and highly innovative in its approach to gain a comprehensive understanding of the optimal pulmonary immune environment for successful control of Mtb infection. It has tremendous potential to generate new insights that will impact the design and implementation of public health interventions in the control of TB in both HIV-uninfected and HIV-infected individuals.
Planned Impact
Current understanding of the immune response underlying successful control of Mtb infections in humans is incomplete. This knowledge gap has contributed, at least in part, to the lack of significant progress in the development of new and effective vaccines against TB. This project will address this problem by exploring new areas of investigation. It is ironic but the HIV-mediated loss of immune control of Mtb infection in the lung actually offers a unique opportunity to probe the mechanistic basis underlying the successful control of TB by airway immune cells. This is significant because it is likely to provide invaluable insights into the nature of the protective immune response that we may wish to induce using interventions such as new TB vaccines. It is my strong belief that to reduce the burden of TB in HIV-infected and HIV-uninfected individuals, it is essential that the relevant mechanisms underlying the pathogenesis of both infections are understood well and taken into account when designing and implementing public health interventions. This project will introduce improve current understanding of immune control of TB in the lung and may contribute important knowledge to ongoing efforts to develop new drugs and vaccines for TB.
Publications
Afran L
(2022)
Defective Monocyte Enzymatic Function and an Inhibitory Immune Phenotype in Human Immunodeficiency Virus-Exposed Uninfected African Infants in the Era of Antiretroviral Therapy.
in The Journal of infectious diseases
Aston SJ
(2019)
Etiology and Risk Factors for Mortality in an Adult Community-acquired Pneumonia Cohort in Malawi.
in American journal of respiratory and critical care medicine
Boliar S
(2019)
Inhibition of the lncRNA SAF drives activation of apoptotic effector caspases in HIV-1-infected human macrophages.
in Proceedings of the National Academy of Sciences of the United States of America
Burke RM
(2021)
Incidence of HIV-positive admission and inpatient mortality in Malawi (2012-2019).
in AIDS (London, England)
Gludish DW
(2020)
TZM-gfp cells: a tractable fluorescent tool for analysis of rare and early HIV-1 infection.
in Scientific reports
Gooden TE
(2022)
Incidence of Cardiometabolic Diseases in People With and Without Human Immunodeficiency Virus in the United Kingdom: A Population-Based Matched Cohort Study.
in The Journal of infectious diseases
Gupta-Wright A
(2020)
Virological failure, HIV-1 drug resistance, and early mortality in adults admitted to hospital in Malawi: an observational cohort study.
in The lancet. HIV
Gupta-Wright A
(2020)
Tuberculosis in Hospitalized Patients With Human Immunodeficiency Virus: Clinical Characteristics, Mortality, and Implications From the Rapid Urine-based Screening for Tuberculosis to Reduce AIDS Related Mortality in Hospitalized Patients in Africa.
in Clinical infectious diseases : an official publication of the Infectious Diseases Society of America
Description | Are HIV-1-infected alveolar macrophages productive sites of viral persistence? |
Amount | $909,300 (USD) |
Funding ID | 4R33AI136097-03 |
Organisation | Cornell University |
Sector | Academic/University |
Country | United States |
Start | 08/2020 |
End | 07/2023 |
Description | Effect of HIV infection on the function of CD8+ polycytotoxic T lymphocytes in human tuberculosis |
Amount | € 444,200 (EUR) |
Funding ID | STE 925/4-1 |
Organisation | German Research Foundation |
Sector | Charity/Non Profit |
Country | Germany |
Start | 03/2021 |
End | 02/2024 |
Description | Federation of African Immunological Society Travel scholarships |
Amount | $3,000 (USD) |
Organisation | Bill and Melinda Gates Foundation |
Sector | Charity/Non Profit |
Country | United States |
Start | 12/2017 |
End | 12/2017 |
Description | Infectious Diseases in Africa Symposium Travel Scholarships |
Amount | $5,000 (USD) |
Organisation | National Institutes of Health (NIH) |
Sector | Public |
Country | United States |
Start | 09/2017 |
End | 09/2017 |
Description | Lung immune function in human TB infection and its perturbation by HIV-1 |
Amount | $3,300,000 (USD) |
Funding ID | 1R01AI155319-01 |
Organisation | Cornell University |
Sector | Academic/University |
Country | United States |
Start | 06/2020 |
End | 05/2025 |
Title | Fluorescent Mycobacterium tuberculosis (Mtb) reporter strains |
Description | We have optimised the use of fluorescent Mtb reporter strains to study the pathogenesis of Mtb in human phagocytic cells. The reporters were developed by our collaborator at Cornell University. |
Type Of Material | Cell line |
Year Produced | 2017 |
Provided To Others? | Yes |
Impact | Many laboratories around the world are using the fluorescent Mtb reporter strains to study the biology of Mtb infections in different animal species and cells. |
Title | Mtb infection of human alveolar macrophages |
Description | We are building a body of data on how different human alveolar macrophage subsets control Mtb infections. The database will link immunological and transcriptomic data with cell function. |
Type Of Material | Database/Collection of data |
Year Produced | 2017 |
Provided To Others? | No |
Impact | Other researcher will use the database to study Mtb infection of other human phagocytes. |
Description | Are HIV-infected alveolar macrophages productive sites of viral persistence? |
Organisation | Cornell University |
Department | College of Veterinary Medicine |
Country | United States |
Sector | Academic/University |
PI Contribution | Collection of human lung samples and performing experiments to determine if the lung is a reservoir of HV-1. |
Collaborator Contribution | They perform single cell RNAseq on samples from Malawi. |
Impact | Yes, this is a multi-disciplinary collaboration involving clinicians, immunologists, computational biologists and cell biologists. |
Start Year | 2020 |
Description | Effect of HIV infection on the function of CD8+ polycytotoxic T lymphocytes in human tuberculosis |
Organisation | University Hospital Ulm |
Country | Germany |
Sector | Hospitals |
PI Contribution | Collecting lung and blood samples and conducting experiments in Malawi for comparison with data from the German site |
Collaborator Contribution | Collection of blood samples and conducting experiments in Germany for comparison with data from the Malawi site |
Impact | Yes, the collaboration is multi-disciplinary involving immunologists, clinicians and microbiologists There are no outputs yet from this collaboration |
Start Year | 2021 |
Description | Infection of human alveolar macrophages using Mtb reporter strains |
Organisation | Cornell University |
Department | Department of Microbiology and Immunology |
Country | United States |
Sector | Academic/University |
PI Contribution | We recruit study participants, process human samples, infect human alveolar macrophages with fluorescent Mycobacterium tuberculosis (Mtb) reporter strains, prepare samples for RNAseq and acquire/analyse flow cytometry data. |
Collaborator Contribution | They donated the fluorescent Mtb strains for use in Malawi. They will perform RNAseq analysis of Mtb-infected and uninfected cells from Malawi and will help with data analysis. |
Impact | A) Technology development: Reporter Bead and Zymosan Assays for assessing phagocyte function; Fluorescent In Situ Hybridisation (FISH) assay for detecting HIV-infected cells B) Publications: 1. Peno C, Banda DH, Jambo N, Kankwatira AM, Malamba R, Allain TJ, Ferreira DM, Heyderman RS, Russell DG, Mwandumba HC, Jambo KC. Alveolar T-Helper 17 responses to Streptococcus pneumoniae are preserved in ART-untreated and treated HIV-infected Malawian adults. J Infect 2017; pii: S0163-4453(17)30343-2, doi: 10.1016/j.jinf.2017.10.013. 2. Gupta-Wright A, Tembo D, Jambo KC, Chimbayo E, Mvaya L, Caldwell S, Russell DG, Mwandumba HC. Functional analysis of phagocyte activity in whole blood from HIV/TB-infected individuals using a novel flow cytometry-based assay. Frontiers Immunol 2017; 8:1222. 3. Jambo KC, Tembo DL, Kamng'ona AW, Musicha P, Banda DH, Kankwatira AM, Malamba RD, Allain TJ, Heyderman RS, Russell DG, Mwandumba HC. HIV-associated disruption of lung cytokines networks is incompletely restored in asymptomatic HIV-infected Malawian adults on antiretroviral therapy. ERJ Open Res 2017; 3(4). pii: 00097-2017. Doi:10.1183/23120541. 4. Wilburn KM, Mwandumba HC, Jambo KC, Boliar S, Solouki S, Russell DG, Gludish DW. Heterogeneous loss of HIV transcription and proviral DNA from 8E5/LAV lymphoblastic leukemia cells revealed by RNA FISH:FLOW analyses. Retrovirology 2016 Aug 11; 13(1):55. doi 10.1186/s12977-016-0289-2. 5. Jambo KC, Banda DH, Afran L, Kankwatira AM, Malamba RD, Allain TJ, Gordon SB, Heyderman RS, Russell DG, Mwandumba HC. Asymptomatic HIV-infected individuals on antiretroviral therapy show impaired lung CD4+ T cell responses to mycobacteria. Am J Respir Crit Care Med 2014; 190:938-47. 6. Jambo KC, Banda DH, Kankwatira AM, Sukumar N, Allain TJ, Heyderman RS, Russell DG, Mwandumba HC. Small alveolar macrophages are infected preferentially by HIV and exhibit impaired phagocytic function. Mucosal Immunol 2014; 7:1116-26 7. Podinovskaia M, VanderVen BC, Yates RM, Glennie S, Fullerton D, Mwandumba HC, and Russell DG. Dynamic, Quantitative Assays of Phagosomal Function. Curr Protocols 2013; 102:14.34.1-14.34.14 8. Russell DG, Vanderven BC, Glennie S, Mwandumba H, Heyderman RS. The macrophage marches on its phagosome: dynamic assays of phagosome function. Nat Rev Immunol 2009; 9:594-600. 9. Mwandumba HC, Squire SB, White SA, Nyirenda MH, Kampondeni DS, Rhoades ER, Zijlstra EE, Molyneux ME, Russell DG. Association between sputum smear status and local immune responses at the site of disease in HIV-infected patients with pulmonary tuberculosis. Tuberculosis 2008; 88:58-63. 10. Mwandumba HC, Squire SB, White SA, Nyirenda MH, Zijlstra EE, Molyneux ME, Russell DG, Rhoades ER. Alveolar macrophages from HIV-infected patients with pulmonary tuberculosis retain the capacity to respond to stimulation by lipopolysaccharide. Microbes Infect 2007; 9:1053-1060. 11. Mwandumba HC, Russell DG, Nyirenda MH, Anderson J, White SA, Molyneux ME, Squire SB. Mycobacterium tuberculosis resides in non-acidified vacuoles in endocytically-competent alveolar macrophages from patients with tuberculosis and HIV infection. J Immunol 2004; 172:4592-8. 12. Russell DG, Mwandumba HC and Rhoades ER. Mycobacterium and the coat of many lipids. J Cell Biol. 2002; 158: 421-426. |
Start Year | 2019 |
Description | Lung immune function in human TB infection and its perturbation by HIV-1 |
Organisation | Cornell University |
Department | College of Veterinary Medicine |
Country | United States |
Sector | Academic/University |
PI Contribution | We collect lung samples and infect them with Mycobacterium tuberculosis to assess the role of host immunity in controlling tuberculosis in the human lung. |
Collaborator Contribution | They infect mice with the same strains of Mycobacterium tuberculosis as we do in Malawi and assess their ability to control infection. |
Impact | Yes, this is a multi-disciplinary collaboration involving clinicians, immunologists, cell biologists, computational biologists and microbiologists. |
Start Year | 2020 |
Description | Radio Programme, Malawi Broadcasting Corporation (MBC) |
Form Of Engagement Activity | A broadcast e.g. TV/radio/film/podcast (other than news/press) |
Part Of Official Scheme? | No |
Geographic Reach | National |
Primary Audience | Media (as a channel to the public) |
Results and Impact | Radio programme on Malawi's biggest national broadcaster to discuss how biomedical research, particularly my award, would benefit patients with tuberculosis and the scientific community. |
Year(s) Of Engagement Activity | 2017 |