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Glycolytic Metabolism of Macrophages Differs by Spatial Location and Subset in Tuberculous Granulomas

Gould, Victoria (2020) Glycolytic Metabolism of Macrophages Differs by Spatial Location and Subset in Tuberculous Granulomas. Master's Thesis, University of Pittsburgh. (Unpublished)

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Abstract

Tuberculosis (TB), caused by infection with Mycobacterium tuberculosis (Mtb), triggers the formation of granulomas in the host. Granulomas are composed of many different types of host immune cells including T and B lymphocytes, macrophages, and neutrophils (PMN). The metabolic pathways used by immune cells in granulomas are important for cell function and glycolytic metabolic pathways in granulomas may be involved limiting or promoting disease. By understanding immunometabolism in TB granulomas, we can improve diagnosis and potentially create new therapeutics to combat TB disease. GLUT1 is a glucose transporter that transports glucose into the cell. To determine what cell subsets express GLUT1 in a granuloma, IHC was performed on lung granuloma-containing tissue sections from non-human primates (NHP) that were experimentally infected with Mtb. These slides were stained for macrophage markers including CD11c and CD163, and GLUT1, before being imaged by fluorescence microscopy. Image analysis was performed using ImageJ to determine the total pixel area and percent pixel area of each cell marker occupied in granuloma cross sections, as well as co-localization between the macrophage markers and GLUT1. To identify if there is a relationship between glucose uptake and mycobacterial antigens, we performed a glucose uptake assay and hypoxic experiments using 2-NBDG on monocyte-derived macrophages that were stimulated with inactivated Mtb. Image analysis revealed co-localization of different cell markers and GLUT1. When looking at co-localization of CD11c and CD163 with GLUT1, we found that CD11c+CD163- epithelioid macrophages, the cells in granulomas that are most commonly infected with Mtb, expressed more GLUT1 than interstitial and alveolar macrophages. Moreover, we see evidence that macrophages increase their glucose (2-NBDG) uptake when stimulated with inactivated Mtb. Identifying immune cells that express GLUT1 and what triggers these immune cells to switch to glycolytic metabolism will help us further understand overall granuloma metabolism and cell differentiation after Mtb infection. In terms of public health, by better understanding the immunometabolism of granulomas, it will improve monitoring of disease progression or to aid in the treatment of Mtb infection to help reduce tuberculosis cases worldwide.


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Details

Item Type: University of Pittsburgh ETD
Status: Unpublished
Creators/Authors:
CreatorsEmailPitt UsernameORCID
Gould, Victoriavag27@pitt.eduvag27
ETD Committee:
TitleMemberEmail AddressPitt UsernameORCID
Thesis AdvisorMattila, Joshuajmattila@pitt.edujmattila
Committee ChairMailliard, Robbierbm19@pitt.edurbm19
Committee ChairAnderson, Carolyncja34@pitt.educja34
Date: 30 July 2020
Date Type: Publication
Defense Date: 26 May 2020
Approval Date: 30 July 2020
Submission Date: 1 April 2020
Access Restriction: No restriction; Release the ETD for access worldwide immediately.
Number of Pages: 63
Institution: University of Pittsburgh
Schools and Programs: Graduate School of Public Health > Infectious Diseases and Microbiology
Degree: MS - Master of Science
Thesis Type: Master's Thesis
Refereed: Yes
Uncontrolled Keywords: Nonhuman primates (NHP) Immunohistochemistry (IHC)
Date Deposited: 31 Jul 2020 02:27
Last Modified: 31 Jul 2020 02:27
URI: http://d-scholarship.pitt.edu/id/eprint/39267

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