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Updated: Aug 8, 2026
Generation of Human Monocyte-derived Dendritic Cells from Whole Blood
Published on: December 24, 2016
Activation of human dendritic cells by bacillus Calmette-Guerin
Insights
Bacillus Calmette-Guerin (BCG) enhances the immune-boosting capabilities of human dendritic cells. BCG treatment increases interleukin-8 (IL-8) production and significantly improves dendritic cells
Area of Science:
- Immunology
- Cell Biology
- Cancer Research
Background:
- Dendritic cells (DCs) are crucial antigen-presenting cells that initiate antitumor immune responses.
- Previous research demonstrated that Bacillus Calmette-Guerin (BCG) can stimulate cultured human dendritic cells.
Purpose of the Study:
- To investigate the expression of interleukin-8 (IL-8) by human dendritic cells stimulated with BCG.
- To evaluate the T cell stimulatory capacity of BCG-treated dendritic cells in mixed leukocyte reactions.
Main Methods:
- Human dendritic cells were generated from peripheral blood mononuclear cells cultured with GM-CSF and IL-4.
- Interleukin-8 (IL-8) protein release was quantified using immunoassay; gene transcription was assessed via RT-PCR.
- T cell proliferation was measured by [3H]thymidine incorporation in mixed leukocyte reactions comparing untreated and BCG-exposed DCs.
Main Results:
- BCG significantly upregulated IL-8 messenger RNA expression and protein secretion in a dose- and time-dependent manner.
- Indomethacin further enhanced BCG-induced IL-8 release.
- BCG-treated dendritic cells exhibited markedly enhanced T cell stimulatory potential compared to untreated cells.
Conclusions:
- BCG treatment boosts the production of IL-8, a key chemokine for T cells and granulocytes, by human dendritic cells.
- BCG enhances the allogeneic T cell stimulatory capacity of human dendritic cells, suggesting a role in immunotherapy.
Purpose:
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.
Materials And Methods:
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.
Results:
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.
Conclusions:
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.
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