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Recurrent Respiratory Papillomatosis (RRP): Increased TH2-Like Chemokine Expression
S104 Abstracts
Asthmatic Children Display Increased Th1 Biased Immunity to Human Metapneumovirus R. N. Douville1, Y. Li2, N. Bastien3, A. Becker4, A. Kozyrskyj5, K. T. HayGlass4; 1Department of Immunology, University of Manitoba/CIHR National Training Program in Allergy and Asthma Research, Winnipeg, MB, CANADA, 2Department of Medical Microbiology, University of Manitoba/Canadian Science Center for Human and Animal, Winnipeg, MB, CANADA, 3Canadian Science Center for Human and Animal Health, Winnipeg, MB, CANADA, 4Depts of Immunology/Medical Microbiology/Pediatrics/Child Health, University of Manitoba/CIHR National Training Program in Allergy and Asthma Research, Winnipeg, MB, CANADA, 5Depts of Immunology/Pediatrics/Child Health/Community Health Sciences, University of Manitoba/CIHR National Training Program in Allergy and Asthma Research, Winnipeg, MB, CANADA. RATIONALE: Severe Respiratory syncytial virus (RSV), and more recently human Metapneumovirus (MPV), infections causing bronchiolitis have a long standing epidemiologic association with asthma pathogenesis. Emerging evidence suggests they may also be triggers of asthma exacerbation. Here, we evaluated whether recall cytokine responses to these viruses associate with current clinical status and a prior history of risk factors for infantile bronchiolitis. METHODS: Peripheral blood mononuclear cells (PBMC) from 8-9 year old children (n>60) were cultured with live RSV and MPV, and the frequency and intensity of type 1 (IFN, CXCL10), type 2 (IL-13), CCL5 and IL-10 virus-specific recall responses in the supernatants were quantified. Clinical parameters, such as physician-diagnosed asthma, skin prick test and airway hyperresponsiveness measurements were related to virusspecific cytokine responses. In addition, virus-specific responses were analysed based on parent-reported history of asthma, exacerbation during colds and infantile bronchitis. RESULTS: Children with self-reported or physician diagnosed asthma demonstrated increased production of Th1 cytokines in response to MPV compared to non-asthmatic children (IFN; p<0.05 and IP-10; p<0.05). This difference in IFN and CXCL10 (type 1 immunity) was not apparent following RSV stimulation. In contrast, asthmatic children displayed weaker CCL5 responses against RSV than non-asthmatic counterparts (CCL5; p<0.05). CONCLUSIONS: Asthmatic children display increased type 1 biased immunity in response to MPV, but not RSV. These respiratory viruses provide a powerful model for examining the impact of viral infections in modulating host immunity in asthmatic vs. non-atopic individuals. SUPPORT: Tom and Mindel Olenick Award in Immunology and Manitoba Health Research Council. Funding: CIHR
405
SUNDAY
Recurrent Respiratory Papillomatosis (RRP): Increased TH2Like Chemokine Expression D. W. Rosenthal1,2, J. A. Devoti1,2, R. Wu3,2, H. Schmidtmayerova4, B. M. Steinberg3,2, V. R. Bonagura1,2; 1Allergy/Immunology, Long Island Jewish Medical Center, Great Neck, NY, 2Institute for Medical Research of North Shore-Long Island Jewish Health System, New Hyde Park, NY, 3Otolaryngology, Long Island Jewish Medical Center, New Hyde Park, NY, 4Institute for Medical Research of North Shore-Long Island Jewish Health System, Manhasset, NY. RATIONALE: RRP is characterized by recurrent benign tumors of the respiratory tract, caused by infection with human papillomavirus 6 and 11 (HPV). The immunologic mechanism(s) that govern disease variation remain unresolved. We hypothesize that CCL18, and other TH2chemokines play a critical role in the immunomodulation of patients with RRP. METHODS: CCL18 was measured in the sera of patients (n=26) and controls (n=18) by DuoSet ELISA. PBMC were exposed to 50 ng/ml of HPV11 E6 for 24 hrs and the supernatants assayed by CCL18 ELISA. HU133 v2.0 microarrays (Affymetrix) were used to compare transcript levels in papilloma tissue with levels in normal adjacent laryngeal tissue from two RRP patients.
406
J ALLERGY CLIN IMMUNOL FEBRUARY 2006
RESULTS: CCL18 was increased in the sera of RRP patients (p<0.0003, n=26). E6-exposed PBMC also expressed increased amounts of CCL18 (n=3 patients). Papillomas showed an increase in the expression of CCL18 (p<0.003), a marked increase in the expression of CCL20 (p<0.0006), and a significant reduction in CCL19 (p<0.0001) and CCL21 (p<0.0001) by microarray. CONCLUSIONS: Increased expression of CCL18 and 20, with a concomitant decrease in CCL19 and 21, would generate a TH2-like milieu that would inhibit TH1-like T-cell responses to HPV. This supports our previous results that expression of IL-4 and the TH1 regulatory cytokineIL-10 are maintained in RRP, with a concomitant decrease in TH1-like cytokine (IFN-, IL-12, and IL-18) expression. We propose that TH2-like immune responses predominate in RRP and thereby permit HPV to evade effective HPV-specific CTL responses, leading to the development of severe disease. Funding: NIH Suppression of Toll-like Receptors 3-, 7- and 9-induced B cell Activation and Antibody Production by Repetitive Oligonucleotides from Mammalian Telomere C. Sackesen1,2, M. Akdis1, J. Zumkehr1, S. S. Alkan3, K. Blaser1, I. Gursel4, C. A. Akdis1; 1Swiss Institute of Allergy and Asthma Research (SIAF), Davos, SWITZERLAND, 2Dept. of Pediatric Allergy and Asthma, Hacettepe University School of Medicine, Ankara, TURKEY, 33M Pharmaceuticals, St. Paul, MN, 4Section of Retroviral Immunology, Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, MD. RATIONALE: Toll-like receptor (TLR)-deriven innate immune response has deleterious consequences such as autoimmunity and septic shock. It has been demonstrated that repetitive TTAGGG elements (sup ODN), present in mammalian telomeres, down-regulate TLR9-mediated immune activation. We investigated the effects of sup ODN on human B cell induced by TLR3 (poly I: C), TLR7 (imidazoquinolone) and TLR9 (CpG2006) ligands. METHODS: Human PBMC and purified B cells were stimulated with specific ligands for TLR2, 3, 4, 5, 7, 8 and 9 in the presence or absence of sup ODN. B cell proliferation, differentiation and antibody-production were determined by [3H]-thymidine incorporation, FACS analysis, ELISA and ELISPOT. RESULTS: Human B cells proliferate and turn into antibody-secreting cells in response to TLR3, 7 and 9 ligands. These ligands similarly induced a program of B cell activation leading to blast formation with increased expression of CD27, CD38, CD86, BCMA with CD19 and CD20. Determination of the frequency of antibody-producing cells revealed that a fraction of activated-B cells turned into particularly IgA-, but also IgG1- and IgG4-producing plasma cells, whereas a high percentage remained as memory-B cells. Sup ODN significantly inhibited the above futures of B cells demonstrating a natural mechanism for the control of B cell memory. CONCLUSIONS: These findings demonstrate that stimulation of human B cells via endosomal associated, but not cell surface expressed TLRs (TLR3, TLR7 and TLR9) plays a pivotal role on polyclonal B cell activation and IgA, IgG1 and IgG4 production as a mechanism of immunological memory. In addition, this work establishes that host-derived telomere ODN can down-regulate B cell responses. Funding: EAACI
407
Role of Dendritic Cells in the Induction of Tolerant Versus Effector T Lymphocytes-Mediated Responses to Amoxicillin R. R-Pena1, S. Lopez1, C. Mayorga1, C. Antunez2, M. J. Torres2, J. A. Cornejo-Garcia1, T. D. Fernandez1, M. Blanca2; 1Research Lab, Carlos Haya Hospital, Malaga, SPAIN, 2Allergy Service, Carlos Haya Hospital, Malaga, SPAIN. RATIONALE: Dendritic cells (DCs) are critical for the development of specific T immune responses, being the only cells able to stimulate naive lymphocytes. DCs are also fundamental for the discrimination between tolerant or effector responses, depending on their differential immature or
408
Asthmatic Children Display Increased Th1 Biased Immunity to Human Metapneumovirus R. N. Douville1, Y. Li2, N. Bastien3, A. Becker4, A. Kozyrskyj5, K. T. HayGlass4; 1Department of Immunology, University of Manitoba/CIHR National Training Program in Allergy and Asthma Research, Winnipeg, MB, CANADA, 2Department of Medical Microbiology, University of Manitoba/Canadian Science Center for Human and Animal, Winnipeg, MB, CANADA, 3Canadian Science Center for Human and Animal Health, Winnipeg, MB, CANADA, 4Depts of Immunology/Medical Microbiology/Pediatrics/Child Health, University of Manitoba/CIHR National Training Program in Allergy and Asthma Research, Winnipeg, MB, CANADA, 5Depts of Immunology/Pediatrics/Child Health/Community Health Sciences, University of Manitoba/CIHR National Training Program in Allergy and Asthma Research, Winnipeg, MB, CANADA. RATIONALE: Severe Respiratory syncytial virus (RSV), and more recently human Metapneumovirus (MPV), infections causing bronchiolitis have a long standing epidemiologic association with asthma pathogenesis. Emerging evidence suggests they may also be triggers of asthma exacerbation. Here, we evaluated whether recall cytokine responses to these viruses associate with current clinical status and a prior history of risk factors for infantile bronchiolitis. METHODS: Peripheral blood mononuclear cells (PBMC) from 8-9 year old children (n>60) were cultured with live RSV and MPV, and the frequency and intensity of type 1 (IFN, CXCL10), type 2 (IL-13), CCL5 and IL-10 virus-specific recall responses in the supernatants were quantified. Clinical parameters, such as physician-diagnosed asthma, skin prick test and airway hyperresponsiveness measurements were related to virusspecific cytokine responses. In addition, virus-specific responses were analysed based on parent-reported history of asthma, exacerbation during colds and infantile bronchitis. RESULTS: Children with self-reported or physician diagnosed asthma demonstrated increased production of Th1 cytokines in response to MPV compared to non-asthmatic children (IFN; p<0.05 and IP-10; p<0.05). This difference in IFN and CXCL10 (type 1 immunity) was not apparent following RSV stimulation. In contrast, asthmatic children displayed weaker CCL5 responses against RSV than non-asthmatic counterparts (CCL5; p<0.05). CONCLUSIONS: Asthmatic children display increased type 1 biased immunity in response to MPV, but not RSV. These respiratory viruses provide a powerful model for examining the impact of viral infections in modulating host immunity in asthmatic vs. non-atopic individuals. SUPPORT: Tom and Mindel Olenick Award in Immunology and Manitoba Health Research Council. Funding: CIHR
405
SUNDAY
Recurrent Respiratory Papillomatosis (RRP): Increased TH2Like Chemokine Expression D. W. Rosenthal1,2, J. A. Devoti1,2, R. Wu3,2, H. Schmidtmayerova4, B. M. Steinberg3,2, V. R. Bonagura1,2; 1Allergy/Immunology, Long Island Jewish Medical Center, Great Neck, NY, 2Institute for Medical Research of North Shore-Long Island Jewish Health System, New Hyde Park, NY, 3Otolaryngology, Long Island Jewish Medical Center, New Hyde Park, NY, 4Institute for Medical Research of North Shore-Long Island Jewish Health System, Manhasset, NY. RATIONALE: RRP is characterized by recurrent benign tumors of the respiratory tract, caused by infection with human papillomavirus 6 and 11 (HPV). The immunologic mechanism(s) that govern disease variation remain unresolved. We hypothesize that CCL18, and other TH2chemokines play a critical role in the immunomodulation of patients with RRP. METHODS: CCL18 was measured in the sera of patients (n=26) and controls (n=18) by DuoSet ELISA. PBMC were exposed to 50 ng/ml of HPV11 E6 for 24 hrs and the supernatants assayed by CCL18 ELISA. HU133 v2.0 microarrays (Affymetrix) were used to compare transcript levels in papilloma tissue with levels in normal adjacent laryngeal tissue from two RRP patients.
406
J ALLERGY CLIN IMMUNOL FEBRUARY 2006
RESULTS: CCL18 was increased in the sera of RRP patients (p<0.0003, n=26). E6-exposed PBMC also expressed increased amounts of CCL18 (n=3 patients). Papillomas showed an increase in the expression of CCL18 (p<0.003), a marked increase in the expression of CCL20 (p<0.0006), and a significant reduction in CCL19 (p<0.0001) and CCL21 (p<0.0001) by microarray. CONCLUSIONS: Increased expression of CCL18 and 20, with a concomitant decrease in CCL19 and 21, would generate a TH2-like milieu that would inhibit TH1-like T-cell responses to HPV. This supports our previous results that expression of IL-4 and the TH1 regulatory cytokineIL-10 are maintained in RRP, with a concomitant decrease in TH1-like cytokine (IFN-, IL-12, and IL-18) expression. We propose that TH2-like immune responses predominate in RRP and thereby permit HPV to evade effective HPV-specific CTL responses, leading to the development of severe disease. Funding: NIH Suppression of Toll-like Receptors 3-, 7- and 9-induced B cell Activation and Antibody Production by Repetitive Oligonucleotides from Mammalian Telomere C. Sackesen1,2, M. Akdis1, J. Zumkehr1, S. S. Alkan3, K. Blaser1, I. Gursel4, C. A. Akdis1; 1Swiss Institute of Allergy and Asthma Research (SIAF), Davos, SWITZERLAND, 2Dept. of Pediatric Allergy and Asthma, Hacettepe University School of Medicine, Ankara, TURKEY, 33M Pharmaceuticals, St. Paul, MN, 4Section of Retroviral Immunology, Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, MD. RATIONALE: Toll-like receptor (TLR)-deriven innate immune response has deleterious consequences such as autoimmunity and septic shock. It has been demonstrated that repetitive TTAGGG elements (sup ODN), present in mammalian telomeres, down-regulate TLR9-mediated immune activation. We investigated the effects of sup ODN on human B cell induced by TLR3 (poly I: C), TLR7 (imidazoquinolone) and TLR9 (CpG2006) ligands. METHODS: Human PBMC and purified B cells were stimulated with specific ligands for TLR2, 3, 4, 5, 7, 8 and 9 in the presence or absence of sup ODN. B cell proliferation, differentiation and antibody-production were determined by [3H]-thymidine incorporation, FACS analysis, ELISA and ELISPOT. RESULTS: Human B cells proliferate and turn into antibody-secreting cells in response to TLR3, 7 and 9 ligands. These ligands similarly induced a program of B cell activation leading to blast formation with increased expression of CD27, CD38, CD86, BCMA with CD19 and CD20. Determination of the frequency of antibody-producing cells revealed that a fraction of activated-B cells turned into particularly IgA-, but also IgG1- and IgG4-producing plasma cells, whereas a high percentage remained as memory-B cells. Sup ODN significantly inhibited the above futures of B cells demonstrating a natural mechanism for the control of B cell memory. CONCLUSIONS: These findings demonstrate that stimulation of human B cells via endosomal associated, but not cell surface expressed TLRs (TLR3, TLR7 and TLR9) plays a pivotal role on polyclonal B cell activation and IgA, IgG1 and IgG4 production as a mechanism of immunological memory. In addition, this work establishes that host-derived telomere ODN can down-regulate B cell responses. Funding: EAACI
407
Role of Dendritic Cells in the Induction of Tolerant Versus Effector T Lymphocytes-Mediated Responses to Amoxicillin R. R-Pena1, S. Lopez1, C. Mayorga1, C. Antunez2, M. J. Torres2, J. A. Cornejo-Garcia1, T. D. Fernandez1, M. Blanca2; 1Research Lab, Carlos Haya Hospital, Malaga, SPAIN, 2Allergy Service, Carlos Haya Hospital, Malaga, SPAIN. RATIONALE: Dendritic cells (DCs) are critical for the development of specific T immune responses, being the only cells able to stimulate naive lymphocytes. DCs are also fundamental for the discrimination between tolerant or effector responses, depending on their differential immature or
408