title: Mycobacterium tuberculosis invades and replicates within type II alveolar cells.
abstract: Although Mycobacterium tuberculosis is assumed to infect primarily alveolar macrophages after being aspirated into the lung in aerosol form, it is plausible to hypothesize that M. tuberculosis can come in contact with alveolar epithelial cells upon arrival into the alveolar space. Therefore, as a first step toward investigation of the interaction between M. tuberculosis and alveolar epithelial cells, we examined the ability of M. tuberculosis to bind to and invade alveolar epithelial cells in vitro. The H37Rv and H37Ra strains of M. tuberculosis were cultured to mid-log phase and used in both adherence and invasion assays. The A549 human type II alveolar cell line was cultured to confluence in RPMI 1640 supplemented with 5% fetal bovine serum, L-glutamine, and nonessential amino acids. H37Rv was more efficient in entering A549 cells than H37Ra, Mycobacterium avium, and Escherichia coli Hb101, and nonpiliated strain (4.7% +/- 1.0% of the initial inoculum in 2 h compared with 3.1% +/- 0.8%, 2.1% +/- 0.9%, and 0.03% +/- 0.0%, respectively). The invasion was more efficient at 37 degrees C than 30 degrees C (4.7% +/- 1.0% compared with 2.3% +/- 0.8%). H37Rv and H37Ra were both capable of multiplying intracellularly at a similar ration over 4 days. Binding was inhibited up to 55.7% by anti-CD51 antibody (antivitronectin receptor), up to 55% with anti-CD29 antibody (beta(1) integrin), and 79% with both antibodies used together. Update of M. tuberculosis H37Rv was microtubule and microfilament dependent. It was inhibited by 6l.4% in the presence of 10 micron colchicine and by 72.3% in the presence of 3 micron cytochalasin D, suggesting two separate pathways for uptake. Our results show that M. tuberculosis is capable of invading type II alveolar epithelial cells and raise the possibility that invasion of alveolar epithelial cells is associated with the pathogenesis of lung infection.
authors.affiliation: Kuzell Institute for Arthritis & Infectious Diseases, California Pacific Medical Center Research Institute, San Francisco, USA.
authors.forename: L E
authors.initials: LE
authors.lastname: Bermudez
authors.forename: J
authors.initials: J
authors.lastname: Goodman
mesh_terms: D000818:Animals; D015703:Antigens, CD / Q000502:physiology; D002460:Cell Line; D003078:Colchicine / Q000494:pharmacology; D015638:Cytochalasin D / Q000494:pharmacology; D006801:Humans; D039564:Integrin alphaV; D019012:Integrin beta1 / Q000502:physiology; D051379:Mice; D009169:Mycobacterium tuberculosis / Q000502:physiology*; D011650:Pulmonary Alveoli / Q000382:microbiology*
chemical_list: D015703:Antigens, CD; D039564:Integrin alphaV; D019012:Integrin beta1; D015638:Cytochalasin D; D003078:Colchicine