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and IL-6 production but not degranulation from murine bone marrow-derived mast cells
Departments of Microbiology & Immunology and Pathology, Dalhousie University, Halifax, Nova Scotia, Canada
Correspondence: Dr. J. S. Marshall, Department of Microbiology and Immunology, Sir Charles Tupper Medical Building, Dalhousie University, Halifax, Nova Scotia B3H 4H7 Canada. E-mail: Jean.Marshall{at}Dal.ca
Mast cells are sentinel cells critical to the initiation of innate
immune and inflammatory responses, particularly at mucosal surfaces. To
fulfill this function they can be activated by several
pathogen-associated stimuli to produce cytokines with or without
concurrent degranulation. We examined the ability of immunostimulatory
DNA sequences including CpG motifs, which are found in increased
quantities in bacterial DNA, to activate mouse bone marrow-derived mast
cells (mBMMC). Mast cells were treated with a range of doses of
CpG-containing oligodeoxynucleotides or control oligodeoxynucleotides
without CpG within their sequence. There was a dose-dependent increase
in the production of both interleukin-6 (IL-6) and tumor necrosis
factor 
(TNF-) by mast cells treated with the CpG-containing
oligodeoxynucleotides. The cytokine levels induced were directly
related to the number of CpG within a given length of sequence.
Treatment with oligonucleotides containing 3CpG induced an eightfold
increase in TNF production over control incubated mast cells. Other
cytokines, including granulocyte-macrophage colony-stimulating factor,
IL-4, interferon-
, and IL-12 were not induced by oligonucleotide
treatment. Neither CpG containing oligodeoxynucleotides nor control
oligodeoxynucleotides induced degranulation of mast cells. Bacterial
DNA from Escherichia coli also induced IL-6 from mBMMC but
neither calf thymus DNA nor methylase-treated E. coli DNA
had such an effect. Examination of the uptake of Texas red-labeled CpG
and non-CpG-containing oligodeoxynucleotides revealed that they were
both similarly taken up by the mBMMC. These results have important
implications for the mechanism by which mast cells respond to bacteria
and for the potential role of mast cells in DNA vaccination.
Key Words: DNA • cytokines • inflammation • bacteria
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