Strains of Mycobacterium tuberculosis differ in affinity for human osteoblasts and alveolar cells in vitro.

Although the lung is the primary site of infection of tuberculosis, Mycobacterium tuberculosis is capable of causing infection at other sites. In 5-10 % such extra-pulmonary tuberculosis is located in bone tissue of the spine. It is unknown whether host or microbial factors are responsible for the site where extra-pulmonary tuberculosis manifests itself. One MDR isolate belonging to strain F28, one susceptible F11 and one isolate each of susceptible, MDR and XDR F15/LAM4/KZN were cultured in Middlebrook 7H9 media. Human osteoblasts (SaOS-2) and human alveolar epithelial cells (A549) were exposed to these different isolates of M. tuberculosis and invasion capacity and intra-cellular multiplication rates were established. Mouse macrophage (MHS) cells exposed to M. tuberculosis H37Rv served as control. The invasion capacity of F15/LAM4/KZN representatives increased with the level of resistance. The F28 MDR strain showed similar invasion capacity as the XDR F15/LAM4/KZN for pulmonary epthelial cells, whilst the fully susceptible F11 strain displayed a propensity for osteoblasts. The differences observed may in part explain why certain strains are able to cause infection at specific extra-pulmonary sites. We postulated that the development of extra-pulmonary tuberculosis depends on the ability of the microbe to pass effectively through the alveolar epithelial lining and its affinity for cells other than those in pulmonary tissue.

Mycobacterium tuberculosis isolates grown under oxygen deprivation invade pulmonary epithelial cells.

Mycobacterium tuberculosis has the ability to adapt to and survive under different environmental conditions, including oxygen deprivation. To better understand the pathogenesis of M. tuberculosis, we studied the invasion of human alveolar (A549) and human bronchial (BBM) epithelial cell lines by M. tuberculosis isolates cultured under oxygen deprivation. We used isolates belonging to the Beijing and F15/LAM4/KZN families, isolates with unique DNA fingerprints and the laboratory strains H37Rv and H37Ra. We determined that: (1) M. tuberculosis bacilli grown under oxygen deprivation invade epithelial cells, (2) the invasion capacity of all 17 isolates differed, and (3) oxygen deprivation influenced the invasion capacity of these isolates. All isolates invaded the A549 more effectively than the BBM cells. Three of the F15/LAM4/KZN isolates, two of which had extensively drug resistance (XDR) profiles, were at least twice as invasive (≥33%) as the most invasive Beijing isolate (15%) (P < 0.05). We conclude that for a more comprehensive understanding of the pathogenesis of M. tuberculosis, studies should include isolates that have been cultured under oxygen deprivation.

Adhesion to and invasion of pulmonary epithelial cells by the F15/LAM4/KZN and Beijing strains of Mycobacterium tuberculosis.

Globally, specific genotypes of Mycobacterium tuberculosis have been shown to dominate in patients, suggesting that these are more successful pathogens. One such genotype, the F15/LAM4/KZN (KZN) family of M. tuberculosis, has predominated in KwaZulu-Natal, South Africa, since the early 1990s. This strain recently evolved from multidrug-resistant to extensively drug-resistant (XDR). The ability of M. tuberculosis strains belonging to the Beijing family, the KZN family, strains with unique DNA fingerprint patterns and laboratory strains (H37Rv and H37Ra) to adhere to and invade a human alveolar (A549) and a human bronchial (BBM) epithelial cell line was investigated. All strains displayed greater adhesion to and invasion of A549 cells as compared to BBM cells. The Beijing and KZN strains combined showed greater adhesion (28 %) than the unique strains (5 %) (P <0.05). The XDR variant of KZN invaded A549 cells more effectively than the other isolates. These results suggest that the successful spread of the Beijing and KZN strains might be related to their interaction with alveolar epithelium.