Peanut oral immunotherapy results in increased antigen-induced regulatory T-cell function and hypomethylation of forkhead box protein 3 (FOXP3).

BACKGROUND The mechanisms contributing to clinical immune tolerance remain incompletely understood. This study provides evidence for specific immune mechanisms that are associated with a model of operationally defined clinical tolerance. OBJECTIVE Our overall objective was to study laboratory changes associated with clinical immune tolerance in antigen-induced T cells, basophils, and antibodies in subjects undergoing oral immunotherapy (OIT) for peanut allergy. METHODS In a phase 1 single-site study, we studied participants (n = 23) undergoing peanut OIT and compared them with age-matched allergic control subjects (n = 20) undergoing standard of care (abstaining from peanut) for 24 months. Participants were operationally defined as clinically immune tolerant (IT) if they had no detectable allergic reactions to a peanut oral food challenge after 3 months of therapy withdrawal (IT, n = 7), whereas those who had an allergic reaction were categorized as nontolerant (NT; n = 13). RESULTS Antibody and basophil activation measurements did not statistically differentiate between NT versus IT participants. However, T-cell function and demethylation of forkhead box protein 3 (FOXP3) CpG sites in antigen-induced regulatory T cells were significantly different between IT versus NT participants. When IT participants were withdrawn from peanut therapy for an additional 3 months (total of 6 months), only 3 participants remained classified as IT participants, and 4 participants regained sensitivity along with increased methylation of FOXP3 CpG sites in antigen-induced regulatory T cells. CONCLUSION In summary, modifications at the DNA level of antigen-induced T-cell subsets might be predictive of a state of operationally defined clinical immune tolerance during peanut OIT.

[1]  T. Nomura,et al.  Functional delineation and differentiation dynamics of human CD4+ T cells expressing the FoxP3 transcription factor. , 2009, Immunity.

[2]  P. Janson,et al.  FOXP3 Promoter Demethylation Reveals the Committed Treg Population in Humans , 2008, PloS one.

[3]  P. Medawar,et al.  ‘Actively Acquired Tolerance’ of Foreign Cells , 1953, Nature.

[4]  C. Martínez-A,et al.  Coordinated Involvement of Mast Cells and T Cells in Allergic Mucosal Inflammation: Critical Role of the CC Chemokine Ligand 1:CCR8 Axis , 2007, The Journal of Immunology.

[5]  P. Chattopadhyay,et al.  Live-cell assay to detect antigen-specific CD4+ T-cell responses by CD154 expression , 2006, Nature Protocols.

[6]  A. Rudensky,et al.  Foxp3 programs the development and function of CD4+CD25+ regulatory T cells , 2003, Nature Immunology.

[7]  S. Durham,et al.  Induction of IL-10+CD4+CD25+ T cells by grass pollen immunotherapy. , 2003, The Journal of allergy and clinical immunology.

[8]  K. Wucherpfennig,et al.  Induction of antigen-specific regulatory T cells in wild-type mice: Visualization and targets of suppression , 2008, Proceedings of the National Academy of Sciences.

[9]  K. Nadeau,et al.  Rapid oral desensitization in combination with omalizumab therapy in patients with cow's milk allergy. , 2011, The Journal of allergy and clinical immunology.

[10]  M. Levings,et al.  Helios+ and Helios− Cells Coexist within the Natural FOXP3+ T Regulatory Cell Subset in Humans , 2013, The Journal of Immunology.

[11]  S. Durham,et al.  Clinical efficacy and immune regulation with peanut oral immunotherapy. , 2009, The Journal of allergy and clinical immunology.

[12]  K. Nouri-Aria.  Foxp3 expressing regulatory T-cells in allergic disease. , 2009, Advances in experimental medicine and biology.

[13]  P. Valent,et al.  The Basophil-Specific Ectoenzyme E-NPP3 (CD203c) as a Marker for Cell Activation and Allergy Diagnosis , 2004, International Archives of Allergy and Immunology.

[14]  N. Hellings,et al.  T-cell-based immunotherapy in multiple sclerosis: induction of regulatory immune networks by T-cell vaccination , 2006, Expert review of clinical immunology.

[15]  S. Tanabe.  Epitope peptides and immunotherapy. , 2007, Current protein & peptide science.

[16]  M. Humbert,et al.  New chemokine targets for asthma therapy , 2005, Current allergy and asthma reports.

[17]  A. Kemper,et al.  A randomized controlled study of peanut oral immunotherapy: clinical desensitization and modulation of the allergic response. , 2011, The Journal of allergy and clinical immunology.

[18]  V. Kuchroo,et al.  Type 1 regulatory T cells (Tr1) in autoimmunity. , 2011, Seminars in immunology.

[19]  K. Maruyama,et al.  Immunosuppressive properties of regulatory T cells generated by incubation of peripheral blood mononuclear cells with supernatants of human RPE cells. , 2012, Investigative ophthalmology & visual science.

[20]  M. Karlsson,et al.  Allergen-responsive CD4+CD25+ Regulatory T Cells in Children who Have Outgrown Cow's Milk Allergy , 2004, The Journal of experimental medicine.

[21]  Shimon Sakaguchi,et al.  The plasticity and stability of regulatory T cells , 2013, Nature Reviews Immunology.

[22]  E. Knol,et al.  Diagnostic Tests Based on Human Basophils: Potentials, Pitfalls and Perspectives , 2006, International Archives of Allergy and Immunology.

[23]  Y. Belkaid,et al.  Expression of Helios, an Ikaros Transcription Factor Family Member, Differentiates Thymic-Derived from Peripherally Induced Foxp3+ T Regulatory Cells , 2010, The Journal of Immunology.

[24]  C. Akdis,et al.  Role of IL-10 in allergen-specific immunotherapy and normal response to allergens. , 2001, Microbes and infection.

[25]  R. Coffman,et al.  Regulatory interactions between CD45RBhigh and CD45RBlow CD4+ T cells are important for the balance between protective and pathogenic cell- mediated immunity , 1994, The Journal of experimental medicine.

[26]  A. Burks,et al.  A randomized, double-blind, placebo-controlled study of milk oral immunotherapy for cow's milk allergy. , 2008, The Journal of allergy and clinical immunology.

[27]  B. Niggemann,et al.  Oral peanut immunotherapy in children with peanut anaphylaxis. , 2010, The Journal of allergy and clinical immunology.

[28]  J. Bousquet,et al.  Double-blind, placebo-controlled immunotherapy with mixed grass-pollen allergoids. III. Efficacy and safety of unfractionated and high-molecular-weight preparations in rhinoconjunctivitis and asthma. , 1989, The Journal of allergy and clinical immunology.

[29]  B. Chipps.  Oral Immunotherapy for Treatment of Egg Allergy in Children , 2013, Pediatrics.

[30]  G. Bannon,et al.  Classic specific immunotherapy and new perspectives in specific immunotherapy for food allergy , 2001, Allergy.

[31]  D. Hepner,et al.  Symposium on the definition and management of anaphylaxis: summary report. , 2005, The Journal of allergy and clinical immunology.

[32]  C. Akdis,et al.  Cord Blood Derived CD4+CD25high T Cells Become Functional Regulatory T Cells upon Antigen Encounter , 2012, PloS one.

[33]  F. Thien,et al.  Induction of T ‘regulatory’ cells by standardized house dust mite immunotherapy: an increase in CD4+CD25+ interleukin‐10+ T cells expressing peripheral tissue trafficking markers , 2004, Clinical and experimental allergy : journal of the British Society for Allergy and Clinical Immunology.

[34]  A. Neunkirchner,et al.  Naturally occurring regulatory T cells: markers, mechanisms, and manipulation , 2012, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[35]  A. Burks,et al.  Oral immunotherapy for food allergy , 2009, Current allergy and asthma reports.

[36]  K. Nakai,et al.  T cell receptor stimulation-induced epigenetic changes and Foxp3 expression are independent and complementary events required for Treg cell development. , 2012, Immunity.

[37]  A. Ganser,et al.  Isolation strategies of regulatory T cells for clinical trials: phenotype, function, stability, and expansion capacity. , 2011, Experimental hematology.

[38]  P. Dash,et al.  Quantitative impact of thymic selection on Foxp3+ and Foxp3− subsets of self-peptide/MHC class II-specific CD4+ T cells , 2011, Proceedings of the National Academy of Sciences.

[39]  M. Humbert,et al.  Chemokine receptor expression on allergen‐specific T cells in asthma and allergic bronchopulmonary aspergillosis , 2007, Allergy.

[40]  Yeonseok Chung,et al.  Conversion of Th2 memory cells into Foxp3+ regulatory T cells suppressing Th2-mediated allergic asthma , 2010, Proceedings of the National Academy of Sciences.

[41]  Shimon Sakaguchi,et al.  Foxp3-dependent and -independent molecules specific for CD25+CD4+ natural regulatory T cells revealed by DNA microarray analysis. , 2006, International immunology.

[42]  S. Mazmanian,et al.  Inducible Foxp3+ regulatory T-cell development by a commensal bacterium of the intestinal microbiota , 2010, Proceedings of the National Academy of Sciences.

[43]  J. Lafaille,et al.  Regulatory T Cells in Immune Tolerance , 2012 .

[44]  A. Jevnikar,et al.  Transgenic rice for allergy immunotherapy. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[45]  M. Reddish,et al.  Cancer-associated MUC1 mucin inhibits human T-cell proliferation, which is reversible by IL-2 , 1998, Nature Medicine.

[46]  C. Akdis,et al.  Epitope-specific T cell tolerance to phospholipase A2 in bee venom immunotherapy and recovery by IL-2 and IL-15 in vitro. , 1996, The Journal of clinical investigation.

[47]  Alf Hamann,et al.  Epigenetic control of FOXP3 expression: the key to a stable regulatory T-cell lineage? , 2009, Nature Reviews Immunology.

[48]  A. Rudensky,et al.  Regulatory T cells: mechanisms of differentiation and function. , 2012, Annual review of immunology.

[49]  D. Robinson,et al.  Chemokine responsiveness of CD4+ CD25+ regulatory and CD4+ CD25− T cells from atopic and nonatopic donors , 2009, Allergy.

[50]  J. Rolland,et al.  Immunotherapy of allergy: anergy, deletion, and immune deviation. , 1998, Current opinion in immunology.

[51]  D. Levy,et al.  Epigenetic Regulation of Foxp3 Expression in Regulatory T Cells by DNA Methylation1 , 2009, The Journal of Immunology.

[52]  TLR ligands of ryegrass pollen microbial contaminants enhance Th1 and Th2 responses and decrease induction of Foxp3hi regulatory T cells , 2013, European journal of immunology.

[53]  C. Akdis,et al.  Specific immunotherapy and turning off the T cell: how does it work? , 2011, Annals of allergy, asthma & immunology : official publication of the American College of Allergy, Asthma, & Immunology.

[54]  T. Jakob,et al.  Birch Pollen Immunotherapy Leads to Differential Induction of Regulatory T Cells and Delayed Helper T Cell Immune Deviation , 2010, The Journal of Immunology.

[55]  Clelia Di Serio,et al.  Coexpression of CD49b and LAG-3 identifies human and mouse T regulatory type 1 cells , 2013, Nature Medicine.

[56]  Joshua Miller,et al.  Favorable effects of alemtuzumab on allospecific regulatory T-cell generation. , 2012, Human immunology.

[57]  S. Durham,et al.  Long-term tolerance after allergen immunotherapy is accompanied by selective persistence of blocking antibodies. , 2011, The Journal of allergy and clinical immunology.

[58]  T. Nomura,et al.  Control of Regulatory T Cell Development by the Transcription Factor Foxp3 , 2002 .

[59]  A. Rudensky,et al.  Extrathymically generated regulatory T cells control mucosal TH2 inflammation , 2012, Nature.

[60]  Cezmi A Akdis,et al.  T regulatory cells in allergy: novel concepts in the pathogenesis, prevention, and treatment of allergic diseases. , 2005, The Journal of allergy and clinical immunology.

[61]  C. Akdis,et al.  Immunological mechanisms of allergen‐specific immunotherapy , 2011, Allergy.

[62]  K. Nadeau,et al.  Changes in antigen-specific T cell number and function during oral desensitization in cow’s milk allergy enabled with omalizumab , 2012, Mucosal Immunology.

[63]  G. Swan,et al.  Asthma Discordance in Twins Is Linked to Epigenetic Modifications of T Cells , 2012, PLoS ONE.

[64]  A. Mcdonald,et al.  CCR8 Expression Identifies CD4 Memory T Cells Enriched for FOXP3+ Regulatory and Th2 Effector Lymphocytes , 2006, The Journal of Immunology.

[65]  H. Weiner,et al.  Regulatory T cell clones induced by oral tolerance: suppression of autoimmune encephalomyelitis. , 1994, Science.

[66]  Marco A Garcia,et al.  Epigenetic modifications and improved regulatory T-cell function in subjects undergoing dual sublingual immunotherapy. , 2012, The Journal of allergy and clinical immunology.

[67]  H. Sampson.  Peanut oral immunotherapy: is it ready for clinical practice? , 2013, The journal of allergy and clinical immunology. In practice.

[68]  C. Akdis,et al.  Mechanisms of allergen-specific immunotherapy , 2012, Clinical and Translational Allergy.

[69]  S. Rankin,et al.  Chemokines in allergic airway disease. , 2003, Current opinion in pharmacology.

[70]  D. Munn,et al.  Ido expression by dendritic cells: tolerance and tryptophan catabolism , 2004, Nature Reviews Immunology.

[71]  C. Akdis,et al.  Immune Responses in Healthy and Allergic Individuals Are Characterized by a Fine Balance between Allergen-specific T Regulatory 1 and T Helper 2 Cells , 2004, The Journal of experimental medicine.

[72]  K. Nadeau,et al.  Basophil CD203c Levels Are Increased at Baseline and Can Be Used to Monitor Omalizumab Treatment in Subjects with Nut Allergy , 2010, International Archives of Allergy and Immunology.