Infection of human monocytes with Mycobacterium bovis BCG induces production of CC-chemokines

Design. CC-chemokines are potent leukocyte activators and chemoattractants, which have an important role in granuloma formation, function critical for the immune responses to mycobacterial infection. This study investigated whether infection of human monocytes with Mycobacterium bovis bacillus Calmette-Guérin (BCG) elicits secretion of RANTES, macrophage inflammatory protein (MIP)-1α and MIP-1β. Methods. RANTES, MIP-1α and MIP-1β synthesis was measured by the presence protein secretion in the cell culture supernatant as determined by enzyme-linked immunosorbent assay. To investigate the mechanism of M. bovis BCG stimulation of RANTES, we carried out inhibition assays with antibodies to CD40 and we used an intracellular calcium chelator BAPTA-AM. Results. Infection of human monocytes with M. bovis BCG induced RANTES, MIP-1α and MIP-1β secretion in a dose-dependent manner. This stimulation of CC-chemokines production was not attributed to LPS contamination. M. bovis-induced RANTES secretion was dependent upon bacterial uptake and on tumor necrosis factor (TNF)-α. Interestingly, the production of RANTES by M. bovis BCG-infected monocytes occurs through a mechanism that requires intracellular calcium and was significantly inhibited (P < 0.05) with antibodies to CD40. Conclusions. These results suggest that the ability of M. bovis BCG to produce CC-chemokines might lead to protection in the acquired immune response of mycobacterial infection and at the same time indicate that M. bovis BCG-induced RANTES secretion is mediated by CD40 and dependent on the intracellular calcium influx.