The Production of Immunoregulatory Cytokines is Localized to the Extrafollicular Area of Human Tonsils

Abstract

The localization and production at the single cell level of 19 different human cytokines, IL-1 alpha, IL-1 beta, IL-1ra, IL-2, IL-3, IL-4, IL-5, IL-6, IL-8, IL-10, IL-13, TNF alpha, TNF beta, IFN gamma, GM-CSF, G-CSF, and TGF beta 1-3, were studied in cryopreserved tonsillar tissue using immunohistochemical staining. The cytokine producing cells, with the exception of IL-1 expressing cells, had a characteristic morphology due to the accumulation of cytokine onto the Golgi organelle. The production of each cytokine was localized to specific compartments in tonsillar tissue sections from children with tonsillar hypertrophy or recurrent tonsillitis in the resting state. Immunoregulatory cytokines such as IL-2, IL-3, IL-4, G-CSF, GM-CSF and TGF beta were produced in the extrafollicular area and entrapped on the cell membranes as well as in pudels in the extracellular matrix surrounding the producer cells. The dominating cytokines both in tissues from recurrent tonsillitis and tonsillar hypertrophy were GM-CSF, G-CSF, and TGF beta 1-3 which were synthezised predominantly in the reticular crypt site. IL-1 alpha, beta and IL-1ra, on the other hand, were localized to the surface and crypt epithelium and to scattered regions in the extrafollicular area. IL-2, IL-6, IFN gamma and IL-10 were found much more often in sections obtained from recurrent tonsillitis tissue compared with those from tonsillar hypertrophy. Reversely, an excessive production of IL-4 was noted in tonsillar hypertrophy compared with that in recurrent tonsillitis. Thus, concomitant production of multiple cytokines was evident with similarities but also differences in cytokine pattern between the two groups studied. The data suggest that T-cell mediated B-cell activation and differentiation take place in the extrafollicular area. Children with recurrent tonsillitis had a higher amount of B-cells and monocytes compared with children with tonsillar hypertrophy. However, the number of CD3, CD4, CD8 or cytoplasmic Ig-positive cells did not differ between the two groups.

Distribution of Dipeptidyl Peptidase IV in Patients with Chronic Tonsillitis

“…DPP IV cell surface expression on tonsillar lymphocytes in extrafollicular areas indicates the involvement of these lymphocytes in the immune response through the modulation of B-cell functions, as well as the synthesis of different cytokines (1). Using the classical histochemical method, we could not find DPP IV expression in lymphoid follicles, although immunofluorescence analysis with CD26 monoclonal antibody demonstrated moderate fluorescence in the germinal centers of some follicles in samples from RT patients.…”

Prime-Boost Vaccination with rBCG/rAd35 Enhances CD8+ Cytolytic T-Cell Responses in Lesions from Mycobacterium Tuberculosis-Infected Primates

“…The Qwin software can detect and separate 16.7 million different colors, which enable sensitive quantification of positive staining and can easily exclude potential background staining from the analysis. This is a well-established quantitative method that has been extensively used primarily in humans (6,7,(14)(15)(16)(17)(18) and NHPs (19,20) for analysis of a wide range of proteins (that is, cell surface proteins; cytoplasmic, granule-associated or nuclear proteins; secreted proteins; and so on) to describe the phenotype and function of different cell types present in tissue.…”

“…Vaccination with rBCG AFRO-1 and rAd35 AERAS-402 induced strong Mtb-specific T-cell responses in peripheral blood of immunized animals (13). Here, we evaluated the protective efficacy of the rBCG/ rAd35 vaccine in tissue biopsies obtained from different organs of animals challenged with virulent Mtb, using a well-established technology on the basis of in situ immunohistology and quantitative computerized image analysis (6,7,(14)(15)(16)(17)(18)(19)(20). Of note, analysis of cells from the peripheral blood requires manipulation in vitro and only allows a small proportion of reactive lymphocytes to be monitored (21) but is nevertheless the principal method used to analyze vaccine-induced immune responses.…”

The immune status in situ of recurrent tonsillitis and idiopathic tonsillar hypertrophy

“…We measured T H -1 immune response in the tonsillar tissue in RT in correlation with other studies (21)(22)(23)(24). This observation was based on differences between local cytokine concentration TNF-α and IL-4.…”