Cytokine profile in minor salivary glands from patients with bronchial asthma Anne Tsicopoulos, MDa, b, Anne Janin, MDc, Hikmat Akoum, PhDa, Catherine.

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Cytokine profile in minor salivary glands from patients with bronchial asthma Anne Tsicopoulos, MDa, b, Anne Janin, MDc, Hikmat Akoum, PhDa, Catherine Lamblin, MDb, Han Vorng, BSca, Qutayba Hamid, MD, PhDd, Andre-Bernard Tonnel, MDa, b, Benoît Wallaert, MDa, b Journal of Allergy and Clinical Immunology Volume 106, Issue 4, Pages 687-696 (October 2000) DOI: 10.1067/mai.2000.109826 Copyright © 2000 Mosby, Inc. Terms and Conditions

Fig. 1 Immunocytochemical pattern of MSG CD4+ T-cell and mast cell infiltration (A) and of IL-4 and IL-5 immunoreactivity (B) in untreated asthmatic subjects (A) , steroid-treated asthmatic subjects (SA) , and control subjects (C) . Results are expressed as the mean number of positive cells ± SEM per field. Statistical analysis was performed by using the Bonferroni t test for multiple comparisons. When differences were statistically significant, P values are indicated. Journal of Allergy and Clinical Immunology 2000 106, 687-696DOI: (10.1067/mai.2000.109826) Copyright © 2000 Mosby, Inc. Terms and Conditions

Fig. 2 Photomicrographs of MSG sections. Example of CD4+ (a , magnification 400×) and tryptase staining (b , magnification 400×; positive cells in red) and of IL-4 protein staining (c , magnification 1000×; positive cells in black) in MSGs from asthmatic patients. Double staining of MSGs from an asthmatic patient with IL-4 and tryptase at 1000× magnification is shown in d . Single IL-4+ cells stain brown-black (B) , single tryptase-positive cells stain red (R) , and double IL-4/tryptase-positive cells stain maroon (M) . MSG section from an asthmatic patient hybridized with an antisense (e , arrows indicate positive cells) and sense (f) probes for IL-4 at 250× magnification is also shown. Positive controls are peripheral T-cell clones from a subject with hyper-IgE syndrome hybridized with an antisense probe for IL-4 (g) and PHA-stimulated mononuclear cells hybridized with an antisense probe for IFN-γ (2 hours) at 400× magnification. Positive cells appear as a well-circumscribed area of silver grains. Journal of Allergy and Clinical Immunology 2000 106, 687-696DOI: (10.1067/mai.2000.109826) Copyright © 2000 Mosby, Inc. Terms and Conditions

Fig. 3 Linear regression analysis of the number of CD4+ cells in MSGs versus FEV1 values in asthmatic patients. Journal of Allergy and Clinical Immunology 2000 106, 687-696DOI: (10.1067/mai.2000.109826) Copyright © 2000 Mosby, Inc. Terms and Conditions

Fig. 4 IL-4 and IFN-γ mRNA expression of MSGs in untreated asthmatic subjects (A) , steroid-treated asthmatic subjects (SA) , and control subjects (C) . Results are expressed as the mean number of positive cells ± SEM per field. Statistical analysis was performed by using the Bonferroni t test for multiple comparisons. When differences were statistically significant, P values are indicated. Journal of Allergy and Clinical Immunology 2000 106, 687-696DOI: (10.1067/mai.2000.109826) Copyright © 2000 Mosby, Inc. Terms and Conditions

Fig. 5 Summary of the data obtained in different mucosal sites in asthmatic patients in the context of lymphocyte recirculation.The mucosal immune system is composed of the lymphoid tissues associated with mucosal surfaces in the respiratory (bronchus-associated lymphoid tissue) and gastrointestinal (gut-associated lymphoid tissue) tracts and other immune mucosal sites, such as salivary glands. The concept of the common mucosal immune system suggests that activated lymphocytes can migrate from one mucosal site to another one. Therefore one can hypothesize that the sensitization of bronchus-associated lymphocytes in asthma may be responsible for the induction of an immune response in other mucosal sites (see diagram; ie, salivary glands [arrow 1] and gut [arrow 2]) . Similarly activated lymphocytes sensitized at the gut level are able to migrate to the bronchial level (like in Crohn’s disease, arrow 3 ). The table indicates the biologic characteristics of bronchi, MSGs, and gut tissue in asthma. An airway-like inflammation has been reported in the gut of asthmatic patients51; an accumulation of lymphocytes and mast cells, but not eosinophils, has been reported in MSGs in asthma (see “Discussion” section for details).15 Journal of Allergy and Clinical Immunology 2000 106, 687-696DOI: (10.1067/mai.2000.109826) Copyright © 2000 Mosby, Inc. Terms and Conditions