Mast cell histamine and cytokine assays.

Abstract

Mast cells are crucial to the development of chronic allergic inflammation and are likely to play a critical role in host defense. In this chapter methodology for histamine and cytokine assays is provided. Crosslinkage of IgE receptor I (Fc epsilonRI) on cord blood-derived mast cells by myeloma IgE and anti-human IgE is used to induce histamine release. Histamine levels were measured in the culture supernatants using an enzyme-linked immunosorbent assay. A human mast cell line (HMC-1), derived from a patient with mast cell leukemia, was activated with interleukin (IL)-1beta to study cytokine production and gene expression. Cytokine gene expression was evaluated by reverse transcriptase polymerase chain reaction and cytokine production was assayed in culture supernatants using an enzyme-linked immunosorbent assay kit.

• Authors: David S Chi, S Matthew Fitzgerald, Guha Krishnaswamy
• Journal: Methods in molecular biology (Clifton, N.J.) (Methods Mol Biol) Vol. 315 Pg. 203-15 ( 2006) ISSN: 1064-3745 [Print] United States
• PMID: 16110160 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't, Research Support, U.S. Gov't, P.H.S.)

Chemical References

• Cytokines
• Interleukin-8
• Receptors, IgE
• Immunoglobulin E
• Histamine

Topics

• Cell Line
• Cytokines (analysis, immunology)
• Enzyme-Linked Immunosorbent Assay
• Fetal Blood (cytology)
• Histamine (analysis, immunology)
• Histamine Release
• Humans
• Immunoglobulin E (immunology)
• Interleukin-8 (metabolism)
• Mast Cells (chemistry, cytology, immunology)
• Receptors, IgE (metabolism)
• Reverse Transcriptase Polymerase Chain Reaction