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Adenosine release from stimulated mast cells.

ABSTRACT: Adenosine release has been documented in lung tissue exposed to hypoxic conditions or antigen challenge. Exogenous adenosine potentiates mediator release from stimulated rat serosal and mouse bone marrow-derived mast cells. To investigate the production and release of adenosine from stimulated mast cells, rat serosal mast cells were purified on metrizamide gradients, sensitized with anti-dinitrophenol IgE for 30 min at 37 degrees C, and challenged in the presence of 1 microM deoxycoformycin with either dinitrophenol-conjugated bovine serum albumin antigen, the calcium ionophore A23187, or compound 48/80. Reactions were terminated by centrifugation, and the supernatants and pellets were assayed for adenosine and ATP content, respectively, by high performance liquid chromatography. The adenosine concentration of the supernatants increased from 0.036 +/- 0.003 nmol per 10(6) cells to 0.049, 0.056, and 0.129 nmol per 10(6) cells 60 sec after challenge with antigen, 48/80, or A23187, respectively. After ionophore stimulation, increased extracellular adenosine was evident by 15 sec, peaked by 60 sec, and remained constant for at least 5 min. A significant decline in stimulated ATP levels was observed within 30 sec, suggesting that the enhanced adenosine concentrations may result from the breakdown of ATP. Cultured mouse bone marrow-derived mast cells under similar conditions also displayed augmented extracellular adenosine levels with A23187 challenge. This endogenous source of adenosine may act locally through a positive feedback mechanism to potentiate immediate hypersensitivity reactions.

SUBMITTER: Marquardt DL

PROVIDER: S-EPMC391886 | biostudies-other | 1984 Oct

REPOSITORIES: biostudies-other

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Adenosine release from stimulated mast cells.

Marquardt D L DL Gruber H E HE Wasserman S I SI

Proceedings of the National Academy of Sciences of the United States of America 19841001 19

Adenosine release has been documented in lung tissue exposed to hypoxic conditions or antigen challenge. Exogenous adenosine potentiates mediator release from stimulated rat serosal and mouse bone marrow-derived mast cells. To investigate the production and release of adenosine from stimulated mast cells, rat serosal mast cells were purified on metrizamide gradients, sensitized with anti-dinitrophenol IgE for 30 min at 37 degrees C, and challenged in the presence of 1 microM deoxycoformycin with ...[more]

PMID: 6435127

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Project description:Mast cells are hematopoietically-derived tissue immune cells that participate in acquired and innate immunity, as well as in inflammation through release of many chemokines and cytokines, especially in response to the pro-inflammatory peptide substance P (SP). Inflammation is critical in the pathogenesis of many diseases, but the trigger(s) is often unknown. We investigated if mast cell stimulation leads to secretion of mitochondrial components and whether these could elicit autocrine and/or paracrine inflammatory effects. Here we show that human LAD2 mast cells stimulated by IgE/anti-IgE or by the SP led to secretion of mitochondrial particles, mitochondrial (mt) mtDNA and ATP without cell death. Mitochondria purified from LAD2 cells and, when mitochondria added to mast cells trigger degranulation and release of histamine, PGD(2), IL-8, TNF, and IL-1?. This stimulatory effect is partially inhibited by an ATP receptor antagonist and by DNAse. These results suggest that the mitochondrial protein fraction may also contribute. Purified mitochondria also stimulate IL-8 and vascular endothelial growth factor (VEGF) release from cultured human keratinocytes, and VEGF release from primary human microvascular endothelial cells. In order to investigate if mitochondrial components could be secreted in vivo, we injected rats intraperiotoneally (ip) with compound 48/80, which mimicks the action of SP. Peritoneal mast cells degranulated and mitochondrial particles were documented by transimission electron microscopy outside the cells. We also wished to investigate if mitochondrial components secreted locally could reach the systemic circulation. Administration ip of mtDNA isolated from LAD2 cells in rats was detected in their serum within 4 hr, indicating that extravascular mtDNA could enter the systemic circulation. Secretion of mitochondrial components from stimulated live mast cells may act as "autopathogens" contributing to the pathogenesis of inflammatory diseases and may be used as targets for novel treatments.

Dataset Information

Adenosine release from stimulated mast cells.

Publications

Adenosine release from stimulated mast cells.

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