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OA-37-733-24 An in vitro intracellular Mycobacterium tuberculosis and HIV-1 co-infection model suitable for imaging-based platforms

Donnellan, Samantha, Ruggiero, A., Pennington, Shaun ORCID:, Thomas, J., Pollakis, G., Biagini, Giancarlo ORCID: and Paxton, W. (2020) 'OA-37-733-24 An in vitro intracellular Mycobacterium tuberculosis and HIV-1 co-infection model suitable for imaging-based platforms' in 51st World Conference on Lung Health of the International Union Against Tuberculosis and Lung Disease, Online, 20 -24 October 2020.

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Background: Mycobacterium tuberculosis (Mtb) is an obligate, intracellular bacillus, causing the human dis- ease Tuberculosis (TB). The Human Immunodeficiency Virus (HIV-1) is a lentivirus that causes AIDS (Acquired Immune Deficiency Syndrome) in humans. Both diseases are among the leading causes of mortality worldwide. Mtb and HIV-1 are classed as hazard group 3 (HG3) pathogens by the UK Advisory Committee for Danger- ous Pathogens, as such they are worked with and stored in containment level 3 (CL3) laboratories. The aim of this work was to develop a safe methodology to co-in- fect macrophage cells with Mtb and HIV-1 suitable for UK CL3 laboratories to monitor pathogen interactions and for drug screening purposes. Design/Methods: We utilised multiple fluorescence- based methodologies to quantify the co-infection. By employing genetically modified Mtb and HIV-1, both carrying fluorescent molecules, we used flow cytometry and microscopy to study levels of infection in the pres- ence or absence of drugs. Much optimisation was required in the development of these methodologies, from cell number, MOI and viral load.
Results: Co-infections were quantified and monitored using different approaches: fluorometry, luciferase, confocal microscopy and flow cytometry. Co-infections were performed in presence or absence of antiretrovi- ral (Efavirenz [EFV]) and/or a first-line TB antibiotic (Rifampicin [RIF]). EFV completely inhibited HIV-1 infection in a single and co-infection setting, and inter- estingly RIF also reduced the HIV-1 infection rate by 12%. RIF reduced Mtb viability by more than 50% in a single infection and by 57% in a co-infection setting. The combination of drugs and presence of both virus and bacteria significantly affected infection rates and pathogen viability.
Conclusions: We have successfully developed a method- ology to co-infect THP-1 cells with HIV-1 pseudo vi- rus particles and Mtb, ensuring host cell viability. Our method can be easily applied to address biological ques- tions concerning the HIV-1/Mtb co-infection process as well as be used for drug screening.

Item Type: Conference or Workshop Item (Speech)
Additional Information: Abstract Only: Published in The International Journal of Tuberculosis and Lung Disease, 'The Abstract Book', Vol 24 No 10 October 2020 Supplement 2
Subjects: WC Communicable Diseases > Infection. Bacterial Infections > Other Bacterial Infections. Zoonotic Bacterial Infections > WC 302 Actinomycetales infections. Mycobacterium infections
WC Communicable Diseases > Virus Diseases > Acquired Immunodeficiency Syndrome. HIV Infections > WC 503 Acquired immunodeficiency syndrome. HIV infections
WF Respiratory System > Tuberculosis > WF 200 Tuberculosis (General)
Faculty: Department: Biological Sciences > Department of Tropical Disease Biology
Related URLs:
Depositing User: Cathy Waldron
Date Deposited: 06 Nov 2020 14:54
Last Modified: 11 Nov 2020 12:38


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