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Volume 12
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10.3390/pathogens12010024
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Article

Measurement of Autophagy Activity Reveals Time-Dependent, Bacteria-Specific Turnover during Mycobacterium tuberculosis Infection

by
Naomi Okugbeni
1,2,
André du Toit
3,
Victoria Cole-Holman
2,
Glynis Johnson
1,
Ben Loos
3 and
Craig Kinnear
1,2,*
1
DSI-NRF Centre of Excellence for Biomedical Tuberculosis Research, US/SAMRC Centre for Tuberculosis Research, Division of Molecular Biology and Human Genetics, Department of Biomedical Sciences, Faculty of Medicine and Health Sciences, Stellenbosch University, Tygerberg 7505, South Africa
2
South African Medical Research Council Genomics Centre, Tygerberg 7505, South Africa
3
Neuro Research Group, Department of Physiological Sciences, Faculty of Sciences, Stellenbosch University, Stellenbosch 7602, South Africa
*
Author to whom correspondence should be addressed.
Pathogens 2023, 12(1), 24; https://doi.org/10.3390/pathogens12010024
Submission received: 17 November 2022 / Revised: 21 December 2022 / Accepted: 22 December 2022 / Published: 23 December 2022
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Abstract

The intracellular pathogen, Mycobacterium tuberculosis (M. tb) uses various mechanisms to evade its killing. One of such is phagosomal damage and cytosolic translocation which is then targeted by the host’s bactericidal autophagy pathway. It is suggested that cytosolic translocation of M. tb is time-dependent, occurring at later time points of 48 to 72 h post-infection. It is, however, not known whether increased autophagic targeting correlates with these time points of infection. We investigated the time-dependent profile of autophagy activity through the course of M. tb infection in mammalian macrophages. Autophagy activity was inferred by the turnover measurement of autophagy markers and M. tb bacilli in THP-1 and RAW 264.7 macrophages. Over a period of 4 to 72 h, we observed highest autophagy turnover at 48 h of infection in M. tb-containing cells. This was evident by the highest turnover levels of p62 and intracellular M. tb. This supports observations of phagosomal damage mostly occurring at this time point and reveal the correlation of increased autophagy activity. The findings support the preservation of autophagy activity despite M. tb infection while also highlighting time-dependent differences in M. tb-infected macrophages. Future studies may explore time-dependent exogenous autophagy targeting towards host-directed anti-tuberculosis therapy.
Keywords: autophagic flux; autophagy turnover; biomarker; immunofluorescence microscopy; LC3B; p62; RAW 264.7; THP-1; tuberculosis; western blot autophagic flux; autophagy turnover; biomarker; immunofluorescence microscopy; LC3B; p62; RAW 264.7; THP-1; tuberculosis; western blot

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MDPI and ACS Style

Okugbeni, N.; du Toit, A.; Cole-Holman, V.; Johnson, G.; Loos, B.; Kinnear, C. Measurement of Autophagy Activity Reveals Time-Dependent, Bacteria-Specific Turnover during Mycobacterium tuberculosis Infection. Pathogens 2023, 12, 24. https://doi.org/10.3390/pathogens12010024

AMA Style

Okugbeni N, du Toit A, Cole-Holman V, Johnson G, Loos B, Kinnear C. Measurement of Autophagy Activity Reveals Time-Dependent, Bacteria-Specific Turnover during Mycobacterium tuberculosis Infection. Pathogens. 2023; 12(1):24. https://doi.org/10.3390/pathogens12010024

Chicago/Turabian Style

Okugbeni, Naomi, André du Toit, Victoria Cole-Holman, Glynis Johnson, Ben Loos, and Craig Kinnear. 2023. "Measurement of Autophagy Activity Reveals Time-Dependent, Bacteria-Specific Turnover during Mycobacterium tuberculosis Infection" Pathogens 12, no. 1: 24. https://doi.org/10.3390/pathogens12010024

APA Style

Okugbeni, N., du Toit, A., Cole-Holman, V., Johnson, G., Loos, B., & Kinnear, C. (2023). Measurement of Autophagy Activity Reveals Time-Dependent, Bacteria-Specific Turnover during Mycobacterium tuberculosis Infection. Pathogens, 12(1), 24. https://doi.org/10.3390/pathogens12010024

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