title: Measurement of T-helper cytokines secreted by cord blood mononuclear cells in response to allergens.
abstract: It has been proposed that in utero factors may predispose towards the development of childhood atopy. To test this hypothesis, it will be necessary to measure T-helper cell (Th) cytokines secreted by human cord blood mononuclear cells (CBMC) stimulated by allergens. However, to date, it has proven impossible to measure allergen-specific CBMC secretion of the key Th cytokine interleukin-4 (IL-4) using conventional sandwich ELISA techniques. We report for the first time the successful measurement of IL-4 secreted by CBMC stimulated by the allergens timothy grass pollen and house dust mite extract. The method is an adaptation of a novel cell-based ELISA (celELISA), which demonstrated an increased (up to 20-fold) sensitivity to detect IL-4. The method is simple, precise, is no more costly than a conventional ELISA, and can identify individuals in a general population whose CBMC exhibit different cytokine biases in response to allergens. The frequency distribution of IL-4 and interferon-gamma (IFN-gamma) CBMC responses to allergens in the general population approximates to a log-normal distribution, which will permit the application of linear regression techniques in the identification of in utero factors which influence Th bias.
authors.affiliation: Department of Medicine and Therapeutics, Institute of Medical Sciences, Foresterhill, Aberdeen, UK.
authors.forename: G
authors.initials: G
authors.lastname: Devereux
authors.forename: A M
authors.initials: AM
authors.lastname: Hall
authors.forename: R N
authors.initials: RN
authors.lastname: Barker
mesh_terms: D000485:Allergens / Q000276:immunology*; D000818:Animals; D018929:Cell Culture Techniques; D002478:Cells, Cultured; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005260:Female; D005312:Fetal Blood / Q000166:cytology; D006801:Humans; D007371:Interferon-gamma / Q000276:immunology / Q000378:metabolism*; D015847:Interleukin-4 / Q000378:metabolism*; D007963:Leukocytes, Mononuclear / Q000166:cytology / Q000276:immunology; D008925:Mites / Q000276:immunology; D006109:Poaceae / Q000276:immunology; D011058:Pollen / Q000276:immunology; D011247:Pregnancy; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D006377:T-Lymphocytes, Helper-Inducer / Q000276:immunology / Q000378:metabolism*
chemical_list: D000485:Allergens; D015847:Interleukin-4; D007371:Interferon-gamma