Mast cell histamine and cytokine assays.
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.
Chi
David S
DS
Department of Internal Medicine, Quillen College of Medicine, East Tennessee State University, Johnson City, TN, USA.
Fitzgerald
S Matthew
SM
Krishnaswamy
Guha
G
D002460:Cell Line; D016207:Cytokines / Q000032:analysis* / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D005312:Fetal Blood / Q000166:cytology; D006632:Histamine / Q000032:analysis* / Q000276:immunology; D006636:Histamine Release; D006801:Humans; D007073:Immunoglobulin E / Q000276:immunology; D016209:Interleukin-8 / Q000378:metabolism; D008407:Mast Cells / Q000737:chemistry* / Q000166:cytology / Q000276:immunology*; D017455:Receptors, IgE / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction
D016207:Cytokines; D016209:Interleukin-8; D017455:Receptors, IgE; D007073:Immunoglobulin E; D006632:Histamine