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A Rationally Designed Microbial Consortium Attenuates Allergic Asthma in a Murine Model
Abstracts AB97
J ALLERGY CLIN IMMUNOL VOLUME 137, NUMBER 2
A Rationally Designed Microbial Consortium Attenuates Allergic Asthma in a Murine Model
Nikole E. Kimes, PhD, Ricardo B. Valladares, PhD, Din L. Lin, PhD, Susan V. Lynch, PhD; University of California, San Francisco. RATIONALE: Lactobacillus supplementation provides partial attenuation of allergic responses in the airway and alters the composition of the gastrointestinal microbiota. Using microbial profiling data from clinical studies, we designed a bacterial consortium based on enriched taxa and functional pathways associated with Lactobacillussupplementation. We hypothesized that intervention using a microbial consortium would provide improved protection against allergic asthma due to the functional synergism of the consortium members. METHODS: To investigate the protective effects of supplementation, C57BL/6 mice were intratracheally sensitized and challenged with cockroach allergen (CRA). The mice were concurrently supplemented with Lactobacillus johnsonii, the microbial consortium, or both over a three-week period. We evaluated allergic responses and potential mechanisms of protection using flow cytometry, qPCR, 16S rRNA gene sequencing, and histology. RESULTS: Supplementation with the microbial consortium and L. johnsoniicombined provided the most robust protection against allergic asthma, including significant decreases in lung mucin hyperplasia and Muc5 gene expression, as well as decreases in Th2-related cytokines. These responses were correlated with systemic increases in IL-17 secreting leukocytes. CONCLUSIONS: The gut microbiota forms a complex functional network that influences both individual microbial members and host immune responses. Here we show that a rationally designed microbial consortium can provide greater attenuation of allergic asthma than an individual probiotic species. Future research will be aimed at characterizing functional interactions and determining the mechanism of improved protection.
317
The Clinical and Immunological Effects of Pru p 3 Slit on Peach and Peanut Tolerance in Patients with Systemic Allergic Reactions
Francisca G omez, MD, PhD1, Gador Bogas, MD2, Miguel Gonzalez3, Paloma Campo, MD, PhD1, Maria Salas, MD, PhD1, J. A. Huertas1, Araceli Diaz-Perales, PhD, Prof4, Domingo Barber, MD5, Marıa J. Rodriguez3, Miguel Blanca, MD, PhD6, Cristobalina Mayorga, PhD7, Marıa Jose Torres, MD, PhD8; 1Allergy Unit, IBIMA-Regional University Hospital of Malaga, Malaga, Spain, 2Allergy Unit, Regional University Hospital of Malaga-IBIMA, UMA, Malaga, Spain, 3Research Laboratory, IBIMA-Regional University Hospital of Malaga, Malaga, Spain, 4Centre for Plant Biotechnology and Genomics (UPM-INIA), Campus de Montegancedo, Pozuelo de Alarcon, Madrid, Spain, 5Universidad San Pablo-CEU, Madrid, Spain, 6Allergy Service, IBIMA-Regional University Hospital of Malaga-UMA, Malaga, Spain, 7Research Laboratory, IBIMA-Regional University Hospital of Malaga-UMA, Malaga, Spain, 8Allergy Unit, IBIMA, Regional University Hospital of Malaga, UMA, Malaga, Spain. RATIONALE: The peach non-specific lipid transfer protein (nsLTP), Pru p 3, is the primary sensitizer in fruits and responsible for severe reactions in the Mediterranean area. Peach allergy is frequently accompanied with other allergies as peanut. Specific immunotherapy to Pru p 3 could affect both peach and peanut tolerance. The aim is evaluate peach and peanut tolerance and immunological changes after one year of Pru p 3 sublingual immunotherapy (SLIT) in patients with systemic allergic reactions to peach and/or peanut. METHODS: Forty-eight peach allergic patients, 36 treated with SLIT and 12 non-treated, were monitored for 12 months. Treated patients were subclassified as peanut allergic (Group A), sensitized (Group B) or tolerant (Group C). SLIT efficacy was evaluated by skin test reactivity and food
tolerance. Immunological changes were evaluated by monitoring sIgE and sIgG4 levels and basophil reactivity. RESULTS: After one year of SLIT, the wheal area in SPT significantly decreased and there was a significant increase in the amount of peach tolerated in treated patients (p<0.001). Patients in Group A showed a significant decrease in peanut SPT wheal area and an increase in amount of peanut tolerated (p<0.001). Immunological changes were observed in treated patients only, with a significant decrease of sIgE and a parallel increase of sIgG4, sIgG4/sIgE and basophil reactivity for both Pru p 3 and Ara h 9. CONCLUSIONS: After one year Pru p 3 SLIT induces both clinical and immunological changes not only for peach but also for other food allergens relevant in the induction of severe reactions such as peanut.
318
Immune Tolerance Induction Following AIT Is Associated with Induction of Circulating CD4+CXCR5+PD-1+FoxP3+ T Follicular Regulatory Cells
Hjh Hanisah Hj Awg Sharif, BHSc, MSc1,2, Rebecca Parkin, BSc1,2, Constance Ito, MSc1,2, Guy Scadding, MRCP2, Stephen R. Durham, MA, MD, FRCP1,2, Mohamed H. Shamji, BSc, MSc, PhD, FAAAAI1,2; 1Immunomodulation and Tolerance Group, Immune Tolerance Network (ITN) Distributed Centre of Excellence for Allergy & Asthma, Allergy & Clinical Immunology Inflammation, Repair and Development National Heart & Lung Institute, Imperial College London, United Kingdom, 2MRC & Asthma UK Centre in Allergic Mechanisms of Asthma, London, United Kingdom. RATIONALE: Circulating T follicular regulatory cells (CD4+CXCR5+PD-1+FoxP3+ Tfr) are a newly identified subset of CD4+T cells with regulatory properties. We hypothesised that Tfr cells are dysregulated in patients with seasonal allergic rhinitis (SAR) compared to non-atopic controls (NA). Furthermore, grass pollen immunotherapy (AIT) is associated with the induction of Tfr cells which may play a role in tolerance induction during AIT. METHODS: In a cross-sectional study of AIT, PBMCs were isolated from SAR (n513), NA (n513) and AIT (subcutaneous, SCIT (n510) and sublingual, SLIT (n58) immunotherapy)-treated group. Circulating CD4+CXCR5+PD-1+FoxP3+ Tfr, CD4+CXCR5+PD-1+FoxP3-T follicular helper (Tfh) cells were enumerated by flow cytometry. Allergen-induced Tfr and Tfh cell proliferation were also assessed. RESULTS: Symptom scores were lower in SCIT (p<0.0001) and SLIT groups (p<0.05) compared to SAR. The frequency of circulating CD4+CXCR5+PD-1+FoxP3+ Tfr cells were lower in SAR compared to NA (median (IQR):13.90(4.42-18.50);p<0.05) and were elevated in SCIT (24.15(13.10-34.24);p<0.05) and SLIT (26.00(20.7036.49);p<0.05). Conventional FoxP3+ Tregs were elevated in SCIT (6.83(4.83-7.92);p<0.01) and SLIT (6.64(5.48-7.92); p<0.001) compared to SAR (4.06(2.50-4.60). We also confirmed that Tfh cells were higher in SAR (86.10(81.50-95.58) compared to SCIT (75.58(65.7686.90);p<0.05), SLIT (74.00 (63.51-79.30);p<0.001) and NA (77.60(49.25-85.15); p<0.05) groups. In addition, Tfh but not Tfr cells showed increased proliferative capacity in response to in vitrograss pollen allergen stimulation. CONCLUSIONS: For the first time, we show that Tfr and Tfh cells are dysregulated in patients with seasonal allergic rhinitis. AIT induces circulating Tfr cells and these cells may potentially be associated with tolerance induction during AIT.
SATURDAY
316
J ALLERGY CLIN IMMUNOL VOLUME 137, NUMBER 2
A Rationally Designed Microbial Consortium Attenuates Allergic Asthma in a Murine Model
Nikole E. Kimes, PhD, Ricardo B. Valladares, PhD, Din L. Lin, PhD, Susan V. Lynch, PhD; University of California, San Francisco. RATIONALE: Lactobacillus supplementation provides partial attenuation of allergic responses in the airway and alters the composition of the gastrointestinal microbiota. Using microbial profiling data from clinical studies, we designed a bacterial consortium based on enriched taxa and functional pathways associated with Lactobacillussupplementation. We hypothesized that intervention using a microbial consortium would provide improved protection against allergic asthma due to the functional synergism of the consortium members. METHODS: To investigate the protective effects of supplementation, C57BL/6 mice were intratracheally sensitized and challenged with cockroach allergen (CRA). The mice were concurrently supplemented with Lactobacillus johnsonii, the microbial consortium, or both over a three-week period. We evaluated allergic responses and potential mechanisms of protection using flow cytometry, qPCR, 16S rRNA gene sequencing, and histology. RESULTS: Supplementation with the microbial consortium and L. johnsoniicombined provided the most robust protection against allergic asthma, including significant decreases in lung mucin hyperplasia and Muc5 gene expression, as well as decreases in Th2-related cytokines. These responses were correlated with systemic increases in IL-17 secreting leukocytes. CONCLUSIONS: The gut microbiota forms a complex functional network that influences both individual microbial members and host immune responses. Here we show that a rationally designed microbial consortium can provide greater attenuation of allergic asthma than an individual probiotic species. Future research will be aimed at characterizing functional interactions and determining the mechanism of improved protection.
317
The Clinical and Immunological Effects of Pru p 3 Slit on Peach and Peanut Tolerance in Patients with Systemic Allergic Reactions
Francisca G omez, MD, PhD1, Gador Bogas, MD2, Miguel Gonzalez3, Paloma Campo, MD, PhD1, Maria Salas, MD, PhD1, J. A. Huertas1, Araceli Diaz-Perales, PhD, Prof4, Domingo Barber, MD5, Marıa J. Rodriguez3, Miguel Blanca, MD, PhD6, Cristobalina Mayorga, PhD7, Marıa Jose Torres, MD, PhD8; 1Allergy Unit, IBIMA-Regional University Hospital of Malaga, Malaga, Spain, 2Allergy Unit, Regional University Hospital of Malaga-IBIMA, UMA, Malaga, Spain, 3Research Laboratory, IBIMA-Regional University Hospital of Malaga, Malaga, Spain, 4Centre for Plant Biotechnology and Genomics (UPM-INIA), Campus de Montegancedo, Pozuelo de Alarcon, Madrid, Spain, 5Universidad San Pablo-CEU, Madrid, Spain, 6Allergy Service, IBIMA-Regional University Hospital of Malaga-UMA, Malaga, Spain, 7Research Laboratory, IBIMA-Regional University Hospital of Malaga-UMA, Malaga, Spain, 8Allergy Unit, IBIMA, Regional University Hospital of Malaga, UMA, Malaga, Spain. RATIONALE: The peach non-specific lipid transfer protein (nsLTP), Pru p 3, is the primary sensitizer in fruits and responsible for severe reactions in the Mediterranean area. Peach allergy is frequently accompanied with other allergies as peanut. Specific immunotherapy to Pru p 3 could affect both peach and peanut tolerance. The aim is evaluate peach and peanut tolerance and immunological changes after one year of Pru p 3 sublingual immunotherapy (SLIT) in patients with systemic allergic reactions to peach and/or peanut. METHODS: Forty-eight peach allergic patients, 36 treated with SLIT and 12 non-treated, were monitored for 12 months. Treated patients were subclassified as peanut allergic (Group A), sensitized (Group B) or tolerant (Group C). SLIT efficacy was evaluated by skin test reactivity and food
tolerance. Immunological changes were evaluated by monitoring sIgE and sIgG4 levels and basophil reactivity. RESULTS: After one year of SLIT, the wheal area in SPT significantly decreased and there was a significant increase in the amount of peach tolerated in treated patients (p<0.001). Patients in Group A showed a significant decrease in peanut SPT wheal area and an increase in amount of peanut tolerated (p<0.001). Immunological changes were observed in treated patients only, with a significant decrease of sIgE and a parallel increase of sIgG4, sIgG4/sIgE and basophil reactivity for both Pru p 3 and Ara h 9. CONCLUSIONS: After one year Pru p 3 SLIT induces both clinical and immunological changes not only for peach but also for other food allergens relevant in the induction of severe reactions such as peanut.
318
Immune Tolerance Induction Following AIT Is Associated with Induction of Circulating CD4+CXCR5+PD-1+FoxP3+ T Follicular Regulatory Cells
Hjh Hanisah Hj Awg Sharif, BHSc, MSc1,2, Rebecca Parkin, BSc1,2, Constance Ito, MSc1,2, Guy Scadding, MRCP2, Stephen R. Durham, MA, MD, FRCP1,2, Mohamed H. Shamji, BSc, MSc, PhD, FAAAAI1,2; 1Immunomodulation and Tolerance Group, Immune Tolerance Network (ITN) Distributed Centre of Excellence for Allergy & Asthma, Allergy & Clinical Immunology Inflammation, Repair and Development National Heart & Lung Institute, Imperial College London, United Kingdom, 2MRC & Asthma UK Centre in Allergic Mechanisms of Asthma, London, United Kingdom. RATIONALE: Circulating T follicular regulatory cells (CD4+CXCR5+PD-1+FoxP3+ Tfr) are a newly identified subset of CD4+T cells with regulatory properties. We hypothesised that Tfr cells are dysregulated in patients with seasonal allergic rhinitis (SAR) compared to non-atopic controls (NA). Furthermore, grass pollen immunotherapy (AIT) is associated with the induction of Tfr cells which may play a role in tolerance induction during AIT. METHODS: In a cross-sectional study of AIT, PBMCs were isolated from SAR (n513), NA (n513) and AIT (subcutaneous, SCIT (n510) and sublingual, SLIT (n58) immunotherapy)-treated group. Circulating CD4+CXCR5+PD-1+FoxP3+ Tfr, CD4+CXCR5+PD-1+FoxP3-T follicular helper (Tfh) cells were enumerated by flow cytometry. Allergen-induced Tfr and Tfh cell proliferation were also assessed. RESULTS: Symptom scores were lower in SCIT (p<0.0001) and SLIT groups (p<0.05) compared to SAR. The frequency of circulating CD4+CXCR5+PD-1+FoxP3+ Tfr cells were lower in SAR compared to NA (median (IQR):13.90(4.42-18.50);p<0.05) and were elevated in SCIT (24.15(13.10-34.24);p<0.05) and SLIT (26.00(20.7036.49);p<0.05). Conventional FoxP3+ Tregs were elevated in SCIT (6.83(4.83-7.92);p<0.01) and SLIT (6.64(5.48-7.92); p<0.001) compared to SAR (4.06(2.50-4.60). We also confirmed that Tfh cells were higher in SAR (86.10(81.50-95.58) compared to SCIT (75.58(65.7686.90);p<0.05), SLIT (74.00 (63.51-79.30);p<0.001) and NA (77.60(49.25-85.15); p<0.05) groups. In addition, Tfh but not Tfr cells showed increased proliferative capacity in response to in vitrograss pollen allergen stimulation. CONCLUSIONS: For the first time, we show that Tfr and Tfh cells are dysregulated in patients with seasonal allergic rhinitis. AIT induces circulating Tfr cells and these cells may potentially be associated with tolerance induction during AIT.
SATURDAY
316