CONCOMITANT IN-VIVO PRODUCTION OF 19 DIFFERENT CYTOKINES IN HUMAN TONSILS

Accumulating data indicate that cytokines, peptides involved in regula tion of both physiological and pathological immune responses, are prod uced predominantly at the site of local antigen stimulation. Cytokine- producing cells were detected at the protein level in human tonsil tis sue obtained from children with recurrent tonsillitis or infectious mo nonucleosis (IM). Concomitant production of 19 different human cytokin es, interleukin-1 alpha (IL-1 alpha), IL-1 beta, IL-1 receptor antagon ist (ra), IL-2, IL-3, IL-4, IL-5, IL-6, IL-8, IL-10, IL-13, granulocyt e-macrophage colony-stimulating factor (GM-CSF), G-CSF, tumour necrosi s factor-alpha (TNF-alpha), TNF-beta, interferon-gamma (IFN-gamma) and transforming growth factor-beta 1-3 (TGF-beta 1-3), was identified at a single-cell level by indirect immunohistochemical staining procedur es and use of carefully selected cytokine-specific antibodies (Ab). Fr esh frozen sections were fixed with 4% paraformaldehyde and permeabili zed by 0.1% saponin treatment, eluting cholesterol from the cell-surfa ce membrane and the Golgi complex. The intracellular localization of a ll cytokines, except IL-1 and IL-1ra, was demonstrated by a characteri stic local cytoplasmic perinuclear configuration in producer cells. In addition, the immunoreactivity for certain cytokines (IL-2, IL-4, IL- 5, G-CSF and GM-CSF) was expressed on the cell membranes and extended over a large extracellular area encompassing the producer cell. Locali zation of the cytokine to the Golgi organelle was established by co-st aining with a monoclonal antibody (mAb) specific to the Golgi complex. Both the extra- and intracellular cytokine staining reactions could b e blocked by preincubation of the cytokine-specific Ab with the corres ponding purified natural or recombinant cytokine. A complex cytokine p attern was established in both groups studied, where most T-helper typ e 1 (Th1) and Th2 lymphokines were expressed in the tonsils but at dif ferent frequencies and localizations. Cells expressing IL-4, IL-5, IL- 10 and IL-13, (Th2 response) were evident at higher frequencies in rec urrent tonsillitis compared to sections from IM, which were associated with a more pronounced IL-2, IFN-gamma and TNF-beta expression.