Induction of IL-10–producing regulatory T cells with TCR diversity by epitope-specific immunotherapy in pollinosis Kei-ichi Yamanaka, MD, Atsushi Yuta,

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Presentation transcript:

Induction of IL-10–producing regulatory T cells with TCR diversity by epitope-specific immunotherapy in pollinosis Kei-ichi Yamanaka, MD, Atsushi Yuta, MD, Masato Kakeda, MD, Ryogen Sasaki, MD, Hiroshi Kitagawa, MD, Esteban C. Gabazza, MD, Kimihiro Okubo, MD, Ichiro Kurokawa, MD, Hitoshi Mizutani, MD Journal of Allergy and Clinical Immunology Volume 124, Issue 4, Pages 842-845.e7 (October 2009) DOI: 10.1016/j.jaci.2009.06.020 Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 1 A, The percentage of forkhead box P3+CD25highCD4+T-cells (naturally occurring regulatory T cells) was unchanged in patients with pollinosis and in those undergoing SIT compared with controls. B, Tr1 (gate 1) and non-Tr1 (gate 2) cells are gated. C, Tr1 cells show equivalent expression of TGF-β mRNA compared to non-Tr1 cells, different from that of IL-10. D, When cells were costained with anti–TGF-β antibody, the percentage of TGF-β–positive cells was similar between Tr1 (gate 1) and non-Tr1 (gate 2). E, CD4+ lymphocyte proliferation was significantly suppressed in PBMCs from patients who received SIT compared with patients with pollinosis. This suppressive function was abolished by the addition of anti–IL-10 neutralizing antibody or anti–IL-10 receptor antibody. F, The percentage of Tr1 cells was significantly decreased in patients with untreated pollinosis compared with normal controls and increased in patients receiving SIT. Journal of Allergy and Clinical Immunology 2009 124, 842-845.e7DOI: (10.1016/j.jaci.2009.06.020) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 2 T-cell receptor complementarity-determining region 3 (CDR3) size spectratyping analysis of a Tr1 population from patients undergoing immunotherapy. A Gaussian distribution of CDR3 lengths was observed in many BV subfamilies, except for oligoclonal expansion in some BV subfamilies, indicating preservation of a highly diverse T-cell receptor repertoire. Journal of Allergy and Clinical Immunology 2009 124, 842-845.e7DOI: (10.1016/j.jaci.2009.06.020) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Phenotypic characterizationof identified Tr1 cells Phenotypic characterizationof identified Tr1 cells. CD4+ T cells were purified from normal PBMCs and cultured with CD3/CD28 antibodies for 8 hours as described in the text. We analyzed the expression levels of skin-homing receptors and chemokine receptors (cutaneous lymphocyte-associated antigen (CLA), a sialyl Lewisx-related epitope expressed on P-selectin glycoprotein ligand 1, CCR6, CCR10, and CCR4), as well as gut (α4 and β7 integrins) and lymph node (CCR7)–homing receptors on CD4+ T cells. Levels of the skin-homing markers CLA, CCR10, and CCR4 were elevated in the Tr1 population compared with the non-Tr1 population. The expression level of β7 integrin was also increased in the Tr1 population, but the levels of the lymph node marker CCR7 were decreased (n = 10, Mann-Whitney test). Journal of Allergy and Clinical Immunology 2009 124, 842-845.e7DOI: (10.1016/j.jaci.2009.06.020) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

T-cell receptor complementarity-determing region 3 (CDR3) size spectratyping analysis of Tr1 or non Tr1-population from SIT-patient, pollen allergy-patient, or normal control. Journal of Allergy and Clinical Immunology 2009 124, 842-845.e7DOI: (10.1016/j.jaci.2009.06.020) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

T-cell receptor complementarity-determing region 3 (CDR3) size spectratyping analysis of Tr1 or non Tr1-population from SIT-patient, pollen allergy-patient, or normal control. Journal of Allergy and Clinical Immunology 2009 124, 842-845.e7DOI: (10.1016/j.jaci.2009.06.020) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

T-cell receptor complementarity-determing region 3 (CDR3) size spectratyping analysis of Tr1 or non Tr1-population from SIT-patient, pollen allergy-patient, or normal control. Journal of Allergy and Clinical Immunology 2009 124, 842-845.e7DOI: (10.1016/j.jaci.2009.06.020) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

T-cell receptor complementarity-determing region 3 (CDR3) size spectratyping analysis of Tr1 or non Tr1-population from SIT-patient, pollen allergy-patient, or normal control. Journal of Allergy and Clinical Immunology 2009 124, 842-845.e7DOI: (10.1016/j.jaci.2009.06.020) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

T-cell receptor complementarity-determing region 3 (CDR3) size spectratyping analysis of Tr1 or non Tr1-population from SIT-patient, pollen allergy-patient, or normal control. Journal of Allergy and Clinical Immunology 2009 124, 842-845.e7DOI: (10.1016/j.jaci.2009.06.020) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

T-cell receptor complementarity-determing region 3 (CDR3) size spectratyping analysis of Tr1 or non Tr1-population from SIT-patient, pollen allergy-patient, or normal control. Journal of Allergy and Clinical Immunology 2009 124, 842-845.e7DOI: (10.1016/j.jaci.2009.06.020) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

T-cell receptor complementarity-determing region 3 (CDR3) size spectratyping analysis of Tr1 or non Tr1-population from SIT-patient, pollen allergy-patient, or normal control. Journal of Allergy and Clinical Immunology 2009 124, 842-845.e7DOI: (10.1016/j.jaci.2009.06.020) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions