Characterization of dendritic cells from human oral mucosa: A new Langerhans' cell type with high constitutive FcϵRI expression Jean-Pierre Allam, MDa,

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Characterization of dendritic cells from human oral mucosa: A new Langerhans' cell type with high constitutive FcϵRI expression Jean-Pierre Allam, MDa, Natalija Novak, MDa, Christian Fuchs, MDb, Sylvia Asen, MDb, Stefaan Bergé, MDc, Thorsten Appel, MDc, Elisabeth Geiger, MDa, Jarema P. Kochan, PhDd, Thomas Bieber, MD, PhDa Journal of Allergy and Clinical Immunology Volume 112, Issue 1, Pages 141-148 (July 2003) DOI: 10.1067/mai.2003.1607 Copyright © 2003 Mosby, Inc. Terms and Conditions

Fig. 1 Analysis of typical LC markers on oLC. Dead cells were excluded by 7AAD staining, and the CD1a+ population was gated manually as demonstrated in 2 representative dot plots. Representative histograms of typical LC markers like Lag (n = 5), Langerin (Lang , n = 7), E-cadherin (E-cad , n = 5), and chemokine receptor 6 (CCR6 , n = 3) on oLC (upper panel) and sLC (lower panel) . RFI of all samples are placed in the upper part of each histogram (mean ± SEM). Journal of Allergy and Clinical Immunology 2003 112, 141-148DOI: (10.1067/mai.2003.1607) Copyright © 2003 Mosby, Inc. Terms and Conditions

Fig. 2 Expression of Fc receptors on oLC. LC were gated as demonstrated in Fig 1. Representative histograms of CD32 (oLC = 49, sLC = 14), CD16 (oLC = 11, sLC = 9), CD64 (oLC = 11, sLC = 9), γ-chain (oLC = 5, sLC = 3), and CD23 (oLC = 60, sLC = 13) of oLC (upper panel) and sLC (lower panel) . RFI of all samples are placed in the upper part of each histogram (mean ± SEM). Journal of Allergy and Clinical Immunology 2003 112, 141-148DOI: (10.1067/mai.2003.1607) Copyright © 2003 Mosby, Inc. Terms and Conditions

Fig. 3 Investigation of FcϵRI on oLC of atopic and nonatopic donors. The results are expressed as mean rFI ± SEM. A, Total FcϵRI of all samples (AS , oLC n = 49, sLC n = 40) and FcϵRI expression of samples from nonatopic (NAT , oLC n = 33, sLC n = 10) and atopic donors (AT , oLC n = 16, sLC n = 4) of oLC (closed bars ) and sLC (open bars ). B , Correlation between FcϵRI expression on oLC and sIgE of all samples (AS , n = 49), nonatopic (NAT , n = 33) and atopic donors (AT , n = 16). C, SurIgE was compared to FcϵRI expression on all samples of oLC (AS , n = 16) and in view of nonatopic status (NAT , n = 12) or atopic status (AT , n = 4). Correlation coefficient (r ) of FcϵRI and surIgE is placed above the bars. Journal of Allergy and Clinical Immunology 2003 112, 141-148DOI: (10.1067/mai.2003.1607) Copyright © 2003 Mosby, Inc. Terms and Conditions

Fig. 4 Detection of unoccupied FcϵRI on oLC. Representative histograms of total FcϵRIα expression compared to unoccupied FcϵRIα expression and detection of surIgE on oLC (n = 5) and sLC (n = 3). Total FcϵRIα was detected by noninhibitory mAb 22E7, unoccupied FcϵRIα by inhibitory mAb 15A5, and surIgE by mAb Dϵ2 directed against the constant region of human IgE. Journal of Allergy and Clinical Immunology 2003 112, 141-148DOI: (10.1067/mai.2003.1607) Copyright © 2003 Mosby, Inc. Terms and Conditions

Fig. 5 Comparison of MHC complexes and costimulatory molecules. oLC (closed bars ) and sLC (open bars ) were gated based on their CD1a expression. The results are expressed as rFI (mean ± SEM). A , Comparison of MHC class I (oLC n = 11, sLC n = 13) and II (oLC n = 56, sLC n = 25) complexes detected on oLC and sLC. B , Expression of CD40 (n = 13), CD80 (oLC n = 37, sLC n = 13), and CD86 (oLC n = 37, sLC n = 13). C, Correlation between CD40 and FcϵRI expression on freshly isolated oLC (n = 13). D, CD83 (n = 8) expression of freshly isolated oLC and sLC. Journal of Allergy and Clinical Immunology 2003 112, 141-148DOI: (10.1067/mai.2003.1607) Copyright © 2003 Mosby, Inc. Terms and Conditions

Fig. 6 Investigation of allogeneic and autologous T-cell response. Results are shown as rSI. A, Allogeneic stimulatory capacity of freshly isolated oLC compared to sLC (n = 5). Mean of T-cell background proliferation (TCB ), 980.2 cpm. B, Autologous stimulatory experiments of freshly isolated oLC and sLC and after 18 hours of culture (n = 3). TCB, 491 cpm. C, Correlation between FcϵRI expression on freshly isolated oLC and autologous stimulatory capacity (n = 9). D, Allogeneic stimulatory experiments of freshly isolated oLC and sLC and after 18 hours of culture (n = 3). TCB, 1238.5 cpm. Journal of Allergy and Clinical Immunology 2003 112, 141-148DOI: (10.1067/mai.2003.1607) Copyright © 2003 Mosby, Inc. Terms and Conditions