Activation of human dendritic cells by bacillus Calmette-Guerin.

Abstract

Dendritic cells are the most potent antigen presenting cells capable of initiating antitumor immune responses. We previously showed that bacillus Calmette-Guerin (BCG) stimulates cultured human dendritic cells. We extended these studies and tested the ability of cultured human dendritic cells to express interleukin IL-8 in response to BCG. We also investigated the T cell stimulatory potential of BCG treated dendritic cells in mixed leukocyte reactions. Dendritic cells were obtained by culturing plastic adherent mononuclear cells from peripheral blood for 6 days in the presence of granulocyte-macrophage colony-stimulating factor and IL-4. Spontaneous and BCG stimulated IL-8 protein release into culture supernatants was measured by a quantitative immunoassay. IL-8 gene transcription was assessed by reverse transcription-polymerase chain reaction. Untreated and BCG exposed dendritic cells were compared as stimulators of allogeneic T cell proliferation, measured as [3H]thymidine incorporation. BCG stimulated IL-8 messenger ribonucleic acid expression and IL-8 protein release. IL-8 secretion occurred in a dose and time dependent fashion. BCG induced IL-8 release was further enhanced in the presence of indomethacin. BCG treated dendritic cells were much more potent T cell stimulators than untreated dendritic cells. These data demonstrate that BCG enhances the production of IL-8, a potent chemokine of T cells and granulocytes, as well as the T cell stimulatory potential of human dendritic cells.