Allergen-driven limiting dilution analysis of human IL-4 and IFN-gamma production in allergic rhinitis and clinically tolerant individuals.
Difficulties in detecting human IL-4 synthesis in antigen-driven primary culture have led to widespread reliance on less physiologic approaches to T cell activation. Although there is general agreement of a Th2-like bias, the precise defects in cytokine responsiveness remain controversial. Analysis of cytokine protein production by fresh, unselected cell populations in response to cognate, antigen-driven stimulation remains an important goal. Here, limiting dilution analysis (LDA) was used to evaluate antigen-stimulated cytokine gene expression by fresh peripheral blood mononuclear cells (PBMC). PBMC from 19 grass pollen sensitive, allergic rhinitis subjects and normal, non-atopic controls were evaluated 1 month after natural reimmunization (the peak of the local grass pollen season). Surprisingly, highly atopic subjects and clinically non-allergic individuals exhibited virtually equivalent antigen-specific, CD4-dependent cytokine production in response to short-term culture with these common environmental antigens. As anticipated, pronounced increases in Th2-like activity were evident in the circulating immune repertoire of grass pollen sensitive individuals, leading to a median ratio of antigen-stimulated IFN-gamma:IL-4 frequencies of 117:1 among normal subjects versus 4:1 among those with allergic rhinitis (Mann-Whitney U-test, P = 0. 00067). This Th2-like bias reflected both a lower frequency of IFN-gamma-producing cells and a markedly increased frequency of IL-4-producing cells in the circulating grass-pollen specific repertoire of atopic donors. The observation that every atopic and normal subject produced IFN-gamma (+/-IL-4) following antigen re-stimulation argues that the decision as to whether allergy or (clinical) tolerance results, hinges not on a genetically determined capacity of whether allergen-reactive T cells can be stimulated in any given individual by chronic exposure to ubiquitous environmental antigens, but on the nature of the cytokine response that comes to dominate that individual's response.
Li
Y
Y
Department of Immunology, University of Manitoba, Winnipeg, Canada.
Simons
F E
FE
Jay
F T
FT
HayGlass
K T
KT
D006801:Humans; D007108:Immune Tolerance / Q000276:immunology*; D007158:Immunologic Techniques; D007371:Interferon-gamma / Q000032:analysis*; D015847:Interleukin-4 / Q000032:analysis*; D006109:Poaceae / Q000276:immunology*; D011058:Pollen / Q000276:immunology*; D006255:Rhinitis, Allergic, Seasonal / Q000276:immunology*; D018418:Th2 Cells / Q000276:immunology
D015847:Interleukin-4; D007371:Interferon-gamma