Lactic acid bacteria inhibit TH2 cytokine production by mononuclear cells from allergic patients Pierre Pochard, PhDab, Philippe Gosset, PhDb, Corinne.

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Lactic acid bacteria inhibit TH2 cytokine production by mononuclear cells from allergic patients Pierre Pochard, PhDab, Philippe Gosset, PhDb, Corinne Grangette, PhDa, Claude Andre, c, André-Bernard Tonnel, PU-PHb, Joël Pestel, PhDb, Annick Mercenier, PhDa Journal of Allergy and Clinical Immunology Volume 110, Issue 4, Pages 617-623 (October 2002) DOI: 10.1067/mai.2002.128528 Copyright © 2002 Mosby, Inc. Terms and Conditions

Fig. 1 LAB reduce IL-4 production and increase IFN-γ production. PBMCs from healthy donors (n = 11) were preincubated for 3 hours (at 2 × 106/mL) with 107 live colony-forming units of 2 gram-positive LAB strains (LAB1 and LAB2) and one gram-negative control bacterium (TG1) and further stimulated with SEA (2 μg/mL). IL-4 (A) and IFN-γ (B) were quantified after 48 hours' incubation by means of specific ELISA. The asterisk indicates significant inhibition (P ≤ .05) of TH2 cytokine production in response to SEA stimulation. Journal of Allergy and Clinical Immunology 2002 110, 617-623DOI: (10.1067/mai.2002.128528) Copyright © 2002 Mosby, Inc. Terms and Conditions

Fig. 2 Influence of the concentration of LAB on their immunomodulatory properties. PBMCs from healthy donors (n = 4) were preincubated for 3 hours (at 2 × 106/mL) with 4 live strains of LAB (LAB1, LAB2, LAB3, and LAB4) at 2 bacteria/cell ratios (1:1 [striped bars] or 10:1 [solid bars] ) before either being stimulated with the SEA superantigen (2 μg/mL) or not. IL-4 (A) , IL-5 (B) , and IFN-γ (C) were quantified in 48-hour supernatants by means of specific ELISA. The asterisk indicates a significant inhibition (P ≤ .05) of TH2 cytokine production in comparison with the SEA-induced response. Journal of Allergy and Clinical Immunology 2002 110, 617-623DOI: (10.1067/mai.2002.128528) Copyright © 2002 Mosby, Inc. Terms and Conditions

Fig. 3 Parameters influencing the production of cytokines. PBMCs from healthy donors (n = 4, at 2 × 106/mL) were stimulated with SEA (2 μg/mL). A, Mononuclear cells were either preincubated for 3 hours with LAB1 (10:1 ratio) before SEA stimulus (□) or stimulated simultaneously with the LAB strain and the superantigen (□), or LAB strains were added 3 hours after sea stimulus (□). B, Mononuclear cells were preincubated for 3 hours with live LAB1 (10:1), heat-killed LAB1 (HK) , or paraformaldehyde-fixed LAB1 (FK) before stimulation with sea. IL-4 concentration was measured at 48 hours by means of ELISA. The asterisk indicates a significant (P ≤ .05) Inhibition of TH2 Cytokine production in comparison with the sea-induced response. Journal of Allergy and Clinical Immunology 2002 110, 617-623DOI: (10.1067/mai.2002.128528) Copyright © 2002 Mosby, Inc. Terms and Conditions

Fig. 4 Inhibition of allergen-induced TH2 cytokine profile. PBMCs from patients allergic to D pteronyssinus (Dpt ; n = 5) were preincubated for 3 hours (at 2 × 106/mL) with 4 strains of LAB (LAB1, LAB2, LAB3, and LAB4) at 2 bacteria/cell ratios (1:1 [striped bars] or 10:1 [solid bars] ) before related allergen-dependent stimulation (1 IR/mL) or not. IL-4 (A) , IL-5 (B) , and IFN-γ (C) production was determined by means of specific ELISA after 48 hours' stimulation. The asterisk indicates a significant (P ≤ .05) inhibition of TH2 cytokine production in comparison with the SEA-induced response. Journal of Allergy and Clinical Immunology 2002 110, 617-623DOI: (10.1067/mai.2002.128528) Copyright © 2002 Mosby, Inc. Terms and Conditions

Fig. 5 Involvement of monocytes in the inhibition of TH2 cytokine production. A, Monocytes (M ; 2 × 105/mL) of healthy donors were preincubated or not in the presence of autologous CD4+ T cells (T ; 2 × 106/mL) for 3 hours with LAB1 (10:1 ratio for LAB/monocytes) before stimulation with SEA (2 μg/mL). B, CD19+ B cells (B) from healthy donors (2 × 105/mL) were preincubated or not in the presence of autologous CD4+ T cells (T ; 2 × 106/mL) for 3 hours with LAB1 (10:1 ratio) before stimulation with SEA (2 μg/mL). IL-4 concentration was determined by means of ELISA after 48 hours' culture. The asterisk indicates a significant (P ≤ .05) inhibition of TH2 cytokine production in comparison with SEA-induced production. Journal of Allergy and Clinical Immunology 2002 110, 617-623DOI: (10.1067/mai.2002.128528) Copyright © 2002 Mosby, Inc. Terms and Conditions

Fig. 6 Involvement of IL-12 and IFN-γ in the LAB anti-TH2 effect. PBMCs from healthy donors (n = 5) were preincubated for 3 hours (at 2 × 106/mL) with LAB1 strain at a bacteria/cell ratio of 10:1 in the presence of neutralizing anti-IL-12 (αIL-12) or anti-IFN-γ (αIFN- γ) antibodies (10 μg/mL) before SEA stimulation (2 μg/mL). In the negative control, IgG2a antibodies were used at the same concentration. IL-4 and IFN-γ concentrations were determined by means of specific ELISA after 48 hours' stimulation. The asterisk indicates a significant (P ≤ .05) difference between cytokine concentrations. Journal of Allergy and Clinical Immunology 2002 110, 617-623DOI: (10.1067/mai.2002.128528) Copyright © 2002 Mosby, Inc. Terms and Conditions