title: Entry and intracellular replication of Mycobacterium tuberculosis in cultured human microvascular endothelial cells.
abstract: Establishment of pulmonary Mycobacterium tuberculosis infection requires evasion of host innate defenses. In the lung alveoli, epithelial cells naturally resist uptake by the inhaled bacilli while macrophages patrol the epithelial surface and phagocytose foreign microbes. Alveolar microvascular endothelial cells, however, have not been examined as a potential point of direct interaction with the bacilli. It has been shown with other bacterial and viral lung pathogens that the lung endothelial cells are not only a point of interaction, but a source for intracellular replication and chronic infection by the pathogen. To investigate if endothelial cells are susceptible to M. tuberculosis infection, we examined attachment, internalization, and intracellular replication of M. tuberculosis bacilli in an immortalized human lung microvascular endothelial cell line (HULEC). By 6 h post-infection, 12% of infecting bacilli were associated with the HULEC monolayer cells. This was twice the association observed following a similar infection with cells from a human foreskin microvascular endothelial cell line (HMEC-1). As measured by survival after the addition of a high extracellular concentration of the aminoglycoside amikacin, approximately one-third of the associated bacilli were internalized and unavailable to the drug in both cell lines. Using electron microscopy, large numbers of bacilli were visible in the vacuoles of HULEC cells after 48 h post-infection; the presence of bacterial septa between adjacent mycobacteria suggests intracellular replication. These in vitro findings support the hypothesis that lung endothelial cells have the potential to participate in in vivo lung infections.
authors.affiliation: Division of Tuberculosis Elimination, National Center for AIDS, STD and TB Prevention, Centers for Disease Control and Prevention, Atlanta, GA 30333, USA.
authors.forename: Parmod K
authors.initials: PK
authors.lastname: Mehta
authors.forename: Russell K
authors.initials: RK
authors.lastname: Karls
authors.forename: Elizabeth H
authors.initials: EH
authors.lastname: White
authors.forename: Edwin W
authors.initials: EW
authors.lastname: Ades
authors.forename: Frederick D
authors.initials: FD
authors.lastname: Quinn
mesh_terms: D001422:Bacterial Adhesion / Q000502:physiology; D001424:Bacterial Infections / Q000382:microbiology; D048708:Cell Growth Processes / Q000502:physiology; D002460:Cell Line; D042783:Endothelial Cells / Q000378:metabolism / Q000382:microbiology* / Q000473:pathology; D006801:Humans; D018799:Intercellular Adhesion Molecule-1 / Q000096:biosynthesis; D008854:Microscopy, Electron / Q000379:methods; D009169:Mycobacterium tuberculosis / Q000254:growth & development / Q000502:physiology*; D011650:Pulmonary Alveoli / Q000098:blood supply* / Q000378:metabolism / Q000473:pathology; D014397:Tuberculosis, Pulmonary / Q000378:metabolism / Q000382:microbiology / Q000473:pathology
chemical_list: D018799:Intercellular Adhesion Molecule-1