Prolonged culture of mast cells with high-glucose medium enhances the Fc epsilon RI-mediated degranulation response and leukotriene C4 production.
BACKGROUND
Mast cell-released chemical mediators such as histamine, leukotriene (LT) C(4) and prostaglandin (PG) D(2) lead to the onset of allergic disorders. ATP provided from glycolysis is essential for histamine release and LTC(4) secretion from mast cells upon Fc epsilon RI cross-linking, indicating that glucose is a primary environmental factor for mast cell activation. In this study, we investigated whether increases in concentrations of glucose in culture media affect the activation of bone marrow-derived mouse mast cells (BMMCs) upon Fc epsilon RI cross-linking.


METHODS
BMMCs were cultured in RPMI-1640 supplemented with varying concentrations (5.5, 11, 16.5, 22, 27.5 and 33 mM) of D-glucose for 3 h, or 1, 3 or 7 days. D-Mannitol was added to the medium containing 5.5 mMD-glucose for osmotic control. After culturing, these cells were sensitized with anti-TNP IgE and then stimulated with TNP-BSA.


RESULTS
We found that long-term culture (7 days) of BMMCs with 33 mMD-glucose increases the Fc epsilon RI-dependent release of beta-hexosaminidase and LTC(4) without affecting surface expression levels of Fc epsilon RI, intracellular ATP levels or calcium signaling. Biochemical analyses demonstrated that Fc epsilon RI-dependent phosphorylation of cytosolic phospholipase A(2) (cPLA(2)) at the Ser505 residue was significantly increased by culturing with 33 mM glucose.


CONCLUSIONS
Taken together, our data suggest that glucose can augment Fc epsilon RI-mediated mast cell activation, particularly the degranulation response and LTC(4) secretion after prolonged culture of mast cells with high-glucose medium. Moreover, it is suggested that increased phosphorylation of cPLA(2) at the Ser505 residue contributes to the enhancement of LTC(4) secretion.
Division of Molecular Cell Immunology and Allergology, Advanced Medical Research Center, Nihon University Graduate School of Medicine, Tokyo, Japan.
Yuko
Y
Kitahata
Satoshi
S
Nunomura
Tadashi
T
Terui
Chisei
C
Ra
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D001094:Arachidonate 5-Lipoxygenase / Q000235:genetics; D000001:Calcimycin / Q000494:pharmacology; D020013:Calcium Signaling / Q000187:drug effects; D015550:Cell Degranulation / Q000187:drug effects*; D002470:Cell Survival / Q000187:drug effects; D051545:Cyclooxygenase 1 / Q000235:genetics; D051546:Cyclooxygenase 2 / Q000235:genetics; D015870:Gene Expression / Q000235:genetics; D005947:Glucose / Q000494:pharmacology*; D007073:Immunoglobulin E / Q000276:immunology / Q000494:pharmacology; D015850:Interleukin-6 / Q000378:metabolism; D017997:Leukotriene C4 / Q000378:metabolism*; D008407:Mast Cells / Q000187:drug effects / Q000502:physiology*; D051379:Mice; D008810:Mice, Inbred C57BL; D054510:Phospholipases A2, Cytosolic / Q000378:metabolism; D010766:Phosphorylation / Q000187:drug effects; D017455:Receptors, IgE / Q000378:metabolism*; D012694:Serine / Q000378:metabolism; D012710:Serum Albumin, Bovine / Q000276:immunology / Q000494:pharmacology; D013997:Time Factors; D001619:beta-N-Acetylhexosaminidases / Q000378:metabolism
D015850:Interleukin-6; D017455:Receptors, IgE; C038601:trinitrophenyl-bovine serum albumin; D012710:Serum Albumin, Bovine; D017997:Leukotriene C4; D007073:Immunoglobulin E; D000001:Calcimycin; D012694:Serine; D000255:Adenosine Triphosphate; D001094:Arachidonate 5-Lipoxygenase; D051545:Cyclooxygenase 1; D051546:Cyclooxygenase 2; D054510:Phospholipases A2, Cytosolic; D001619:beta-N-Acetylhexosaminidases; D005947:Glucose