Transcriptional Profile of Mycobacterium tuberculosis in an in vitro Model of Intraocular Tuberculosis
Intraocular tuberculosis (IOTB), an extrapulmonary manifestation of tuberculosis of the eye, has unique and varied clinical presentations with poorly understood pathogenesis. As it is a significant cause of inflammation and visual morbidity, particularly in TB endemic countries, it is essential to s...
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Published in | Frontiers in cellular and infection microbiology Vol. 8; p. 330 |
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Main Authors | , , , , , , , |
Format | Journal Article |
Language | English |
Published |
Switzerland
Frontiers Media S.A
02.10.2018
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Subjects | |
Online Access | Get full text |
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Summary: | Intraocular tuberculosis (IOTB), an extrapulmonary manifestation of tuberculosis of the eye, has unique and varied clinical presentations with poorly understood pathogenesis. As it is a significant cause of inflammation and visual morbidity, particularly in TB endemic countries, it is essential to study the pathogenesis of IOTB. Clinical and histopathologic studies suggest the presence of
in retinal pigment epithelium (RPE) cells.
A human retinal pigment epithelium (ARPE-19) cell line was infected with a virulent strain of
(H37Rv). Electron microscopy and colony forming units (CFU) assay were performed to monitor the
adherence, invasion, and intracellular replication, whereas confocal microscopy was done to study its intracellular fate in the RPE cells. To understand the pathogenesis, the transcriptional profile of
in ARPE-19 cells was studied by whole genome microarray. Three upregulated
transcripts were also examined in human IOTB vitreous samples.
Scanning electron micrographs of the infected ARPE-19 cells indicated adherence of bacilli, which were further observed to be internalized as monitored by transmission electron microscopy. The CFU assay showed that 22.7 and 8.4% of the initial inoculum of bacilli adhered and invaded the ARPE-19 cells, respectively, with an increase in fold CFU from 1 dpi (0.84) to 5dpi (6.58). The intracellular bacilli were co-localized with lysosomal-associated membrane protein-1 (LAMP-1) and LAMP-2 in ARPE-19 cells. The transcriptome study of intracellular bacilli showed that most of the upregulated transcripts correspond to the genes encoding the proteins involved in the processes such as adherence (e.g.,
and
), invasion (e.g.,
and
), virulence (e.g.,
and
), and intracellular survival (e.g.,
and
as well as regulators of various metabolic pathways. Two of the upregulated transcripts (
) were also present in the vitreous samples of the IOTB patients.
is phagocytosed by RPE cells and utilizes these cells for intracellular multiplication with the involvement of late endosomal/lysosomal compartments and alters its transcriptional profile plausibly for its intracellular adaptation and survival. The findings of the present study could be important to understanding the molecular pathogenesis of IOTB with a potential role in the development of diagnostics and therapeutics for IOTB. |
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Bibliography: | ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 Reviewed by: Brian Weinrick, Trudeau Institute, United States; Raul G. Barletta, University of Nebraska-Lincoln, United States; Josephine Clark-Curtiss, University of Florida, United States Present Address: Sudhanshu Abhishek, Department of Pediatrics-Infectious Diseases, Center for Infection and Inflammation Imaging Research, Johns Hopkins University School of Medicine, Baltimore, MD, United States This article was submitted to Molecular Bacterial Pathogenesis, a section of the journal Frontiers in Cellular and Infection Microbiology Edited by: Adel M. Talaat, University of Wisconsin-Madison, United States |
ISSN: | 2235-2988 2235-2988 |
DOI: | 10.3389/fcimb.2018.00330 |