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Cytokine Profiles in Monosensitized and Polysensitized Allergic Rhinitis Patients Treated with Sublingual Immunotherapy
AB262 Abstracts
855
Cytokine Profiles in Monosensitized and Polysensitized Allergic Rhinitis Patients Treated with Sublingual Immunotherapy
L. Maslova1, Leonid P. Titov, MD, PhD2, Lawrence M. DuBuske, MD, FAAAAI3,4; 1Medical Academy of Postgraduate Education, Minsk, Belarus, 2Republican Research and Practical Center for Epidemiology and Microbiology, Minsk, Belarus, 3Immunology Research Institute of New England, Gardner, MA, 4George Washington University School of Medicine, Washington, DC. RATIONALE: Cytokines induce allergic inflammation leading to respiratory allergy. This study assesses intracellular cytokine profiles in patients receiving Sublingual immunotherapy (SLIT). METHODS: 60 adult patients with allergic rhinitis 19 to 46 years old, who received SLIT for two years with standardized allergen extracts (Sevapharma) were studied. Group 1 included 30 patients receiving monotherapy with a mixture of grasses I or Artemisia. Group 2 included 30 patients, receiving combination therapy of a mixture of grasses I or Artemisia and mixture of house dust mites or indoor moulds. The control group included 30 healthy subjects. CD4+T-cells were assayed for intracellular cytokines after stimulation with PMA plus ionomycin. RESULTS: CD4+T-cells (IL-4+ cells, IL-5+ cells, IL-13+ cells, or IL-17+ cells) were significantly increased before SLIT compared to controls but were reduced after the 2nd year of SLIT. IFN-g+ cells increased after the 2nd year of SLIT. Comparing CD4+cells before and after SLIT in patients of Group 1, IL-4+ was reduced from 0.91 to 0.3; IL-17+ reduced from 0.75 to 0.38; IL-5+ reduced from 4.71 to 2.05; IL-13+ reduced from 4.9 to 1.58 and IFN-g+ cells increased from 13.37 to 18.85. In Group 2, IL-4+ cells were reduced from 0.75 to 0.28; IL-17+ reduced from 0.5 to 0.2; IL-5+ reduced from 6.11 to 1.88; IL-13+ reduced from 4.95 to 2.06 and IFN-g+ increased from 14.53 to 18.65. CONCLUSIONS: Reduction of IL-4+, IL-17+, IL-5+ and IL-13+ CD4+T-cells and increase of IFN-g+ CD4+T-cells demonstrated that SLIT can modulate immune responses in mono- and poly-sensitized patients.
856
Validation and Verification of Grass Allergen Challenge in the Allergen Biocube (ABC)
MONDAY
Endri Angjeli1, Keith Lane2, Emily Schoemmell3, Yesha Raval3, Paul Gomes4; 1Ora Inc, Andover, MA, 2Ora Inc., MA, 3Ora Inc, 4Ora Inc. RATIONALE: Grass allergy can be difficult to study in field trial due to unreliable exposure patterns and poor correlations between skin test response and severity of nasal symptoms. To overcome these challenges, the Allergen BioCube (ABC) was validated for the delivery of Timothy Grass allergen in a uniform manner to a capacity of 25 subjects inducing a clinically meaningful rhinitis response. METHODS: Twenty five subjects were screened and 14 were enrolled with SPT wheel > 5mm, and no symptoms at the screening Visit 1. Visits 2, 3, 4 and 5 consisted of 3 hour exposures to 4000+/- 450 grains per cubic meter of timothy pollen in consecutive days. PNIF, PEFR, serum specific IGE and clinician graded Nasal Inflammation Score (NIS) were captured. The study was conducted out of season. RESULTS: The average baseline Total Nasal Symptom Score (TNSS) of 0.36 increased to 6.7 62.7 after three hours at Visit 2. This response persisted at Visit 3, 4, and 5 with averages of 6.2 63.4, 5.5 63.5, and 5.5 63.5, respectively. From this population 50% of subjects reached a TNSS > 6 by Visit 5. Similarly the NIS increased to a grade of 3.7 61.6 by Visit 5. CONCLUSIONS: The Allergen BioCube creates a clinically meaningful level of rhinitis in a controlled environment which can be used to evaluate the efficacy of therapies without the concern of a poor natural season and within shorter timelines.
J ALLERGY CLIN IMMUNOL FEBRUARY 2016
857
Contrast Agent Symptoms
Reduces
Allergic
Rhinitis
Erik Viirre, MD, PhD1, J. Ernest Villafranca, PhD1, S. David Miller, MD2, Paul Gomes3, Elliott Lasser, MD1; 13E Therapeutics Corporation, La Jolla, CA, 2North-East Medical Research Associates, 3Ora Inc. RATIONALE: We hypothesized that the contrast agent iodixanol, as Nasapaque nasal solution, was effective in reducing nasal allergy symptoms. METHODS: This clinical trial enrolled 73 adult subjects with seasonal allergic rhinitis and positive ragweed skin tests. After priming at earlier visits, treatment efficacy (Nasapaque or placebo) was assessed at Visit 4 (90 minutes ragweed exposure in Ora’s Allergen BioCubeÒ [ABC]) followed by treatment and additional 7.5 hours ABC exposure) and at Visit 5 (treatment 30 minutes before 3 hours ABC exposure). The primary efficacy endpoint was total nasal symptom score (TNSS, 0-12 scale) assessed at multiple time points pre- and during exposure. Mean TNSS differences between active and placebo treatment groups and change from baseline/ pre-treatment differences were calculated (1-sided t-test, alpha 5 0.10). RESULTS: Subjects treated with Nasapaque (N 5 36) had lower TNSS scores and greater change from baseline than placebo (N 5 37) subjects. Onset of action for Nasapaque was as early as 15 minutes (mean D from baseline -3.2; treatment difference -0.9, 80% CI -1.6 to -0.2, p 5 0.0602). Statistically significant differences were seen as late as 4.25 hours posttreatment (mean D from baseline -4.1; treatment difference -0.9, 80% CI -1.7, -0.1, p 5 0.0754). Visit 4 3-hour post-treatment mean TNSS scores were 3.9 (Nasapaque) and 5.0 (placebo); -1.1 difference, 80% CI -2.0, -0.02, p 5 0.0625. Visit 5 results also indicated greater efficacy with Nasapaque treatment, which was safe and well tolerated. CONCLUSIONS: Nasapaque nasal solution is effective in reducing nasal allergy symptoms. Further evaluations of efficacy are warranted.
858
Three Complementary Pathways Characterize the Suppressive Properties of Epit-Induced Tregs
Benjamin Pelletier, Master degree1, Lucie Mondoulet, PhD1, Emilie Puteaux1, Melanie Ligouis1, Veronique Dhelft1, Camille Plaquet1, Christophe Dupont, MD, PhD2, Pierre-HenriBenhamou,MD1;1DBV Technologies, Bagneux, France, 2Hopital Necker Enfants Malades, Paris, France. RATIONALE: Epicutaneous immunotherapy (EPIT) demonstrates clinical efficacy and induces significant Foxp3+ Tregs in mice. In addition, adoptive transfer of EPIT-induced Tregs protects mice from anaphylaxis and further sensitizations (bystander effect). Nevertheless, mechanisms of action of EPITinduced Tregs are not clearly elucidated. This study analyzed the suppressive properties of EPIT-induced Tregs in specific/bystander conditions. METHODS: Milk-sensitized BALB/c mice were treated or not with milk EPIT. Tregs (CD4+CD25+ T cells) from EPIT, Sham or non-sensitized groups and effector T cells (CD4+CD25-) from milk or peanut sensitized mice were sorted and co-cultured for 4 days at different ratios, using allergen stimulation and allergen-pulsed CD11c+ antigen-presenting cells. Anti CTLA-4, antiTGF-b antibodies were also tested to determine whether EPIT-induced Tregs act via cytokines or by cell-contact dependent mediation. Suppression was analyzed by tracking divided CD4+CD25- with CFSE by flow cytometry. Supernatants were also collected to quantify cytokine secretion. RESULTS: Effector T cells proliferate up to 82.5%-92.5%, respectively with milk or peanut stimulation. Presence of EPIT-induced Tregs significantly inhibited effector T cells proliferation in specific or bystander conditions (68-71% of proliferation, i.e. 20%-25% proliferation inhibition) compared to Sham or non-sensitized Tregs. Interestingly, blocking CTLA-4 and TGF-b strongly abrogated suppression capacity of EPIT-induced Tregs in both conditions. IL-2 and TH2 cytokines were barely detectable in supernatants with EPIT Tregs. CONCLUSIONS: EPIT-induced Tregs inhibited effector T cell proliferation with the same potency in specific and bystander conditions. Suppression induced by EPIT-induced Tregs might use 3 complementary pathways: a low availability of IL-2, TGF-b secretion and CTLA-4 cell contact mediation.
855
Cytokine Profiles in Monosensitized and Polysensitized Allergic Rhinitis Patients Treated with Sublingual Immunotherapy
L. Maslova1, Leonid P. Titov, MD, PhD2, Lawrence M. DuBuske, MD, FAAAAI3,4; 1Medical Academy of Postgraduate Education, Minsk, Belarus, 2Republican Research and Practical Center for Epidemiology and Microbiology, Minsk, Belarus, 3Immunology Research Institute of New England, Gardner, MA, 4George Washington University School of Medicine, Washington, DC. RATIONALE: Cytokines induce allergic inflammation leading to respiratory allergy. This study assesses intracellular cytokine profiles in patients receiving Sublingual immunotherapy (SLIT). METHODS: 60 adult patients with allergic rhinitis 19 to 46 years old, who received SLIT for two years with standardized allergen extracts (Sevapharma) were studied. Group 1 included 30 patients receiving monotherapy with a mixture of grasses I or Artemisia. Group 2 included 30 patients, receiving combination therapy of a mixture of grasses I or Artemisia and mixture of house dust mites or indoor moulds. The control group included 30 healthy subjects. CD4+T-cells were assayed for intracellular cytokines after stimulation with PMA plus ionomycin. RESULTS: CD4+T-cells (IL-4+ cells, IL-5+ cells, IL-13+ cells, or IL-17+ cells) were significantly increased before SLIT compared to controls but were reduced after the 2nd year of SLIT. IFN-g+ cells increased after the 2nd year of SLIT. Comparing CD4+cells before and after SLIT in patients of Group 1, IL-4+ was reduced from 0.91 to 0.3; IL-17+ reduced from 0.75 to 0.38; IL-5+ reduced from 4.71 to 2.05; IL-13+ reduced from 4.9 to 1.58 and IFN-g+ cells increased from 13.37 to 18.85. In Group 2, IL-4+ cells were reduced from 0.75 to 0.28; IL-17+ reduced from 0.5 to 0.2; IL-5+ reduced from 6.11 to 1.88; IL-13+ reduced from 4.95 to 2.06 and IFN-g+ increased from 14.53 to 18.65. CONCLUSIONS: Reduction of IL-4+, IL-17+, IL-5+ and IL-13+ CD4+T-cells and increase of IFN-g+ CD4+T-cells demonstrated that SLIT can modulate immune responses in mono- and poly-sensitized patients.
856
Validation and Verification of Grass Allergen Challenge in the Allergen Biocube (ABC)
MONDAY
Endri Angjeli1, Keith Lane2, Emily Schoemmell3, Yesha Raval3, Paul Gomes4; 1Ora Inc, Andover, MA, 2Ora Inc., MA, 3Ora Inc, 4Ora Inc. RATIONALE: Grass allergy can be difficult to study in field trial due to unreliable exposure patterns and poor correlations between skin test response and severity of nasal symptoms. To overcome these challenges, the Allergen BioCube (ABC) was validated for the delivery of Timothy Grass allergen in a uniform manner to a capacity of 25 subjects inducing a clinically meaningful rhinitis response. METHODS: Twenty five subjects were screened and 14 were enrolled with SPT wheel > 5mm, and no symptoms at the screening Visit 1. Visits 2, 3, 4 and 5 consisted of 3 hour exposures to 4000+/- 450 grains per cubic meter of timothy pollen in consecutive days. PNIF, PEFR, serum specific IGE and clinician graded Nasal Inflammation Score (NIS) were captured. The study was conducted out of season. RESULTS: The average baseline Total Nasal Symptom Score (TNSS) of 0.36 increased to 6.7 62.7 after three hours at Visit 2. This response persisted at Visit 3, 4, and 5 with averages of 6.2 63.4, 5.5 63.5, and 5.5 63.5, respectively. From this population 50% of subjects reached a TNSS > 6 by Visit 5. Similarly the NIS increased to a grade of 3.7 61.6 by Visit 5. CONCLUSIONS: The Allergen BioCube creates a clinically meaningful level of rhinitis in a controlled environment which can be used to evaluate the efficacy of therapies without the concern of a poor natural season and within shorter timelines.
J ALLERGY CLIN IMMUNOL FEBRUARY 2016
857
Contrast Agent Symptoms
Reduces
Allergic
Rhinitis
Erik Viirre, MD, PhD1, J. Ernest Villafranca, PhD1, S. David Miller, MD2, Paul Gomes3, Elliott Lasser, MD1; 13E Therapeutics Corporation, La Jolla, CA, 2North-East Medical Research Associates, 3Ora Inc. RATIONALE: We hypothesized that the contrast agent iodixanol, as Nasapaque nasal solution, was effective in reducing nasal allergy symptoms. METHODS: This clinical trial enrolled 73 adult subjects with seasonal allergic rhinitis and positive ragweed skin tests. After priming at earlier visits, treatment efficacy (Nasapaque or placebo) was assessed at Visit 4 (90 minutes ragweed exposure in Ora’s Allergen BioCubeÒ [ABC]) followed by treatment and additional 7.5 hours ABC exposure) and at Visit 5 (treatment 30 minutes before 3 hours ABC exposure). The primary efficacy endpoint was total nasal symptom score (TNSS, 0-12 scale) assessed at multiple time points pre- and during exposure. Mean TNSS differences between active and placebo treatment groups and change from baseline/ pre-treatment differences were calculated (1-sided t-test, alpha 5 0.10). RESULTS: Subjects treated with Nasapaque (N 5 36) had lower TNSS scores and greater change from baseline than placebo (N 5 37) subjects. Onset of action for Nasapaque was as early as 15 minutes (mean D from baseline -3.2; treatment difference -0.9, 80% CI -1.6 to -0.2, p 5 0.0602). Statistically significant differences were seen as late as 4.25 hours posttreatment (mean D from baseline -4.1; treatment difference -0.9, 80% CI -1.7, -0.1, p 5 0.0754). Visit 4 3-hour post-treatment mean TNSS scores were 3.9 (Nasapaque) and 5.0 (placebo); -1.1 difference, 80% CI -2.0, -0.02, p 5 0.0625. Visit 5 results also indicated greater efficacy with Nasapaque treatment, which was safe and well tolerated. CONCLUSIONS: Nasapaque nasal solution is effective in reducing nasal allergy symptoms. Further evaluations of efficacy are warranted.
858
Three Complementary Pathways Characterize the Suppressive Properties of Epit-Induced Tregs
Benjamin Pelletier, Master degree1, Lucie Mondoulet, PhD1, Emilie Puteaux1, Melanie Ligouis1, Veronique Dhelft1, Camille Plaquet1, Christophe Dupont, MD, PhD2, Pierre-HenriBenhamou,MD1;1DBV Technologies, Bagneux, France, 2Hopital Necker Enfants Malades, Paris, France. RATIONALE: Epicutaneous immunotherapy (EPIT) demonstrates clinical efficacy and induces significant Foxp3+ Tregs in mice. In addition, adoptive transfer of EPIT-induced Tregs protects mice from anaphylaxis and further sensitizations (bystander effect). Nevertheless, mechanisms of action of EPITinduced Tregs are not clearly elucidated. This study analyzed the suppressive properties of EPIT-induced Tregs in specific/bystander conditions. METHODS: Milk-sensitized BALB/c mice were treated or not with milk EPIT. Tregs (CD4+CD25+ T cells) from EPIT, Sham or non-sensitized groups and effector T cells (CD4+CD25-) from milk or peanut sensitized mice were sorted and co-cultured for 4 days at different ratios, using allergen stimulation and allergen-pulsed CD11c+ antigen-presenting cells. Anti CTLA-4, antiTGF-b antibodies were also tested to determine whether EPIT-induced Tregs act via cytokines or by cell-contact dependent mediation. Suppression was analyzed by tracking divided CD4+CD25- with CFSE by flow cytometry. Supernatants were also collected to quantify cytokine secretion. RESULTS: Effector T cells proliferate up to 82.5%-92.5%, respectively with milk or peanut stimulation. Presence of EPIT-induced Tregs significantly inhibited effector T cells proliferation in specific or bystander conditions (68-71% of proliferation, i.e. 20%-25% proliferation inhibition) compared to Sham or non-sensitized Tregs. Interestingly, blocking CTLA-4 and TGF-b strongly abrogated suppression capacity of EPIT-induced Tregs in both conditions. IL-2 and TH2 cytokines were barely detectable in supernatants with EPIT Tregs. CONCLUSIONS: EPIT-induced Tregs inhibited effector T cell proliferation with the same potency in specific and bystander conditions. Suppression induced by EPIT-induced Tregs might use 3 complementary pathways: a low availability of IL-2, TGF-b secretion and CTLA-4 cell contact mediation.