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B-Dependent Network1
Department of Infectious Diseases, Imperial College of Science Technology and Medicine, Hammersmith Campus, London, United Kingdom
Pulmonary epithelial cells, covering a 70-m2 surface
area, have not previously been considered an important source of
chemokines in pulmonary tuberculosis. We analyzed IL-8 secretion from
A549 cells and primary normal human bronchial epithelial cells (NHBE)
infected by Mycobacterium tuberculosis. Direct infection
of A549 cells by M. tuberculosis caused IL-8 secretion
of 7720 ± 1610 pg/106 cells, but no IL-8 secretion
from NHBE after 24 h. In contrast, conditioned media from
M. tuberculosis-infected human monocytes (CoMTB) induced
a much greater IL-8 secretion of 92,635 ± 13,180
pg/106 A549 cells and 13,416 ± 3,529
pg/106 NHBE after 24 h. CoMTB induced rapid IL-8 mRNA
accumulation, which was stable over 24 h, compared with
TNF-
-induced transcripts. CoMTB stimulated nuclear binding of p65,
p50, and c-Rel subunits of NF-
B to IL-8 promoter sequences.
Transient transfections with IL-8 promoter reporter constructs showed
NF-B binding-site mutations abolished IL-8 promoter activity while
NF-IL-6 binding-site mutations decreased promoter activity to 50.2
± 6.3% of wild-type activity. IL-1R antagonist but not neutralizing
anti-TNF- inhibited epithelial cell IL-8 secretion, mRNA
accumulation, and NF-B binding. Recombinant IL-1ß (2 ng/ml)
induced similar levels of IL-8 secretion to CoMTB in both A549 cells
and NHBE. Pulmonary epithelial cells are a major source of IL-8 in the
initial host response to pulmonary tuberculosis. Such IL-8 secretion is
NF-B dependent, NF-IL-6 requiring, and activated by an IL-1-mediated
pathway as a consequence of phagocytosis of M.
tuberculosis by monocytes.
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