Modulation of alveolar macrophage leukotriene B4 released by complement component C5.

The release of neutrophil chemotactic activity by the guinea pig alveolar macrophage (AM) is dependent on the fifth component of complement (C5) on the cell surface. Because one potent chemotactic factor released by AMs is leukotriene B4 (LTB4), we hypothesized that cell surface C5 may modulate LTB4 release. To test this hypothesis, human AMs obtained by bronchoalveolar lavage from 12 subjects were cultured for 4 hours in the presence of anti-C5 Fab' antibodies with stimuli. The cultures were harvested and evaluated for LTB4 by radioimmunoassay. The LTB4 levels in supernatants obtained from AMs cultured in media alone were variable (447 +/- 63 pg/ml), but the levels were increased when AMs were cultured with the stimuli-opsonized zymosan, immune complexes, or lipopolysaccharide (233%, 49%, and 114% increase, respectively, compared with macrophages cultured in media alone, p less than 0.05). Culturing the AMs with anti-C5 Fab' antibodies inhibited the release of LTB4 induced by opsonized zymosan, immune complexes, or lipopolysaccharide (78%, 41%, and 82% inhibition, respectively, p less than 0.05). Consistent with these observations, anti-C5 Fab' antibodies also decreased the neutrophil chemotactic activity of culture supernatants obtained from AMs stimulated with the same stimuli (p less than 0.001). These data suggest that AM release of LTB4 may be C5-dependent.

Bronchial epithelial cells release chemotactic activity for lymphocytes.

Lymphocytes can frequently be observed in association with bronchial tissues. One mechanism that might account for this association is that bronchial epithelial cells might release chemotactic factors for lymphocytes. To test this hypothesis, bovine bronchial epithelial cells were cultured in serum-free media, and the supernatant fluids were harvested and evaluated for lymphocyte chemotactic activity using a blind-well chamber technique. Media alone attracted few lymphocytes (12 +/- 2 cells/high power field), but in contrast, there was a significant increase in the number of cells attracted by supernatant fluids obtained from bronchial epithelial cell cultures (40 +/- 6 cells/high power field, P = 0.002). The activity was dose dependent and was demonstrated to be chemotactic activity by checkerboard analysis. Partial characterization of the activity revealed it was not extractable into ethyl acetate but was partially inactivated by trypsin and heat (100 degrees C, 15 min). The responding cells were predominantly T-helper lymphocytes as shown by monoclonal antibody staining, with a smaller proportion being B-lymphocytes. Molecular sieve column chromatography revealed multiple peaks of lymphocyte chemotactic activity, with three of the peaks preferentially attracting T-helper lymphocytes and one of the peaks preferentially attracting B-lymphocytes. These data demonstrate that bronchial epithelial cells can release chemotactic factors for lymphocytes and suggest that bronchial epithelial cells may modulate their local population of immune effector cells.