Figure 5. Enhanced inflammation in Mcpt5Cre A20^F/F mice in the HDM, IL-33, and OVA + IL-33 asthma models.

(A) Scatter plots show absolute BAL fluid cell numbers of Eos (eosinophils, SSC^hi, Siglec-F⁺, Gr-1^int, CD11c⁻), B cells (FSC/SSC^lo, CD19⁺, MHCII⁺), and T cells (FSC/SSC^lo, CD3⁺, MHCII⁻), as identified by flow cytometric analysis. Cytokine production by MLN cells and HDM-specific IgE levels were measured by ELISA. Bars indicate means, and data are representative of three independent experiments with four to six mice per group (Control, 5 “PBS” and 6 “HDM” Cre⁻ littermates). (B) Mice received 100 µg HDM extract and 10 µg OVA-AF647 intratracheally and were analyzed 18 h later. Scatter plots show absolute numbers of DCs (DCs, CD11c⁺, MHCII^hi, nonautofluorescent) and antigen positive DCs (AF647⁺) in the lung and MLNs as identified by flow cytometric analysis. Bars indicate means from six to seven mice per group (Control, 6 “PBS” and 7 “HDM” Cre⁻ littermates). (C) Representative confocal images of trachea whole mounts of HDM sensitized and challenged mice after i.v. injection of FITC⁺ 500 nm microbeads: green, FITC⁺ microbeads; red, anti-CD31. (D) Mice were administered 100 ng IL-33 i.n. on 3 consecutive days. Scatter plots show absolute lung cell numbers and DCs (CD11c⁺, MHCII^hi, nonautofluorescent) as identified by flow cytometric analysis. Bars indicate means from three to six mice per group (Control, 3 “PBS” and 5 “IL-33” Cre⁻ littermates). (E) Scatter plots show absolute BAL fluid cell numbers, granulocytes (SSC^int→hi, Ly6C⁺, Ly6G^int→hi, CD11b⁺, CD11c⁻), lymphocytes (FSC/SSC^lo, CD3⁺, or CD19⁺), and DCs (CD11c⁺, MHCII^hi, nonautofluorescent) as identified by flow cytometric analysis. Bars indicate means, and data are integrated from two independent experiments with two to six mice per group (Control, 5 “OVA” and 10 “OVA + IL-33” Cre⁻ littermates). *p<0.05, **p<0.01 (Mann-Whitney test).