title: Antigen mediated and polyclonal stimulation of human cytokine production elicit qualitatively different patterns of cytokine gene expression.
abstract: Polyclonal activators are widely used as surrogate antigens in analysis of human cytokine gene expression. An implicit assumption is that the T cell activation and cytokine production observed in response to polyclonal activation provides a more intense, but qualitatively identical, reflection of results that would be obtained with antigen. Here we demonstrate that stimulation using accessory cell independent (immobilized anti-CD3 mAb) or dependent [phytohemagglutinin (PHA) or soluble anti-CD3 mAb] polyclonal activators yields different conclusions from those that are obtained in response to antigen-specific T cell activation. Cytokine synthesis in 1-5 day bulk cultures of fresh peripheral blood mononuclear cells (PBMC) from 52 subjects evenly divided between grass pollen sensitive allergic rhinitis subjects and normal, non-atopic controls were examined. Antigen-specific re-stimulation elicited elevated IL-4 and IL-10 production and lower IFN-gamma synthesis among allergic subjects than normal non-atopic control subjects. This commitment of fresh PBMC towards a Th2-like response in atopics and the dominance of the IFN-gamma response seen in non-allergic subjects was reinforced when the ratio of IFN-gamma:IL-4 production in bulk culture was examined. Atopic individuals exhibited median IFN-gamma:IL-4 values of 0.07, whereas grass pollen stimulated cytokine production by normal subjects yielded a ratio of 4.8. In marked contrast, IL-2, IL-4, IL-10 and IFN-gamma production elicited using polyclonal activators, though much more intense, did not differ between allergic and non-allergic subjects (Wilcoxon rank sum test P > > 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)
authors.affiliation: Department of Immunology, University of Manitoba, Winnipeg, Canada.
authors.forename: M
authors.initials: M
authors.lastname: Imada
authors.forename: F E
authors.initials: FE
authors.lastname: Simons
authors.forename: F T
authors.initials: FT
authors.lastname: Jay
authors.forename: K T
authors.initials: KT
authors.lastname: HayGlass
mesh_terms: D000293:Adolescent; D000328:Adult; D002478:Cells, Cultured; D016207:Cytokines / Q000096:biosynthesis* / Q000235:genetics*; D000939:Epitopes / Q000276:immunology*; D005260:Female; D005786:Gene Expression Regulation / Q000276:immunology; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000276:immunology; D008297:Male; D010835:Phytohemagglutinins / Q000276:immunology*; D006255:Rhinitis, Allergic, Seasonal / Q000276:immunology
chemical_list: D016207:Cytokines; D000939:Epitopes; D010835:Phytohemagglutinins