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Th2 cytokines
S116 Abstracts
Association Between Second-Hand Smoke Exposure (SSE) and Enhanced Peripheral Blood Mononuclear Cell (PMNC) IL-13 Production During Infancy D. Gentile, A. Patel, D. Skoner; Department of Pediatrics, Allegheny General Hospital, Pittsburgh, PA. RATIONALE: IL-13 production has been documented in children and adults with asthma and atopy. SSE is recognized as a risk factor for the development of childhood asthma. The purpose of this pilot study was to determine whether there is an association between SSE and PMNC IL-13 production during infancy. METHODS: SSE was assessed by questionnaire, and blood was obtained at 2 weeks and 5 months of age in 51 healthy infants. PMNC were cultured and stimulated with PHA, and supernatants were assayed for IL-13 by EIA. RESULTS: Twenty-three infants had no history of SSE, and 28 infants had a history of SSE. IL-13 production was significantly increased in infants with SSE as compared to those without SSE at both ages (p<0.05). In those with and without SSE, the mean (± SEM) levels of IL-13 production (ng/ml) were 1.4 + 0.4 and 8.0 + 0.3 at 2 weeks of age, and 1.8 + 0.5 and 8.8 + 0.2 at 5 months of age, respectively. CONCLUSIONS: These results demonstrate an association between SSE and enhanced PMNC IL-13 production during infancy. Future studies need to expand upon these sample sizes and explore whether enhanced PMNC IL-13 production is the mechanism by which SSE predisposes to the development of asthma/atopy. Funding: NIH
463
SUNDAY
Deficient GM-CSF and Flt-3 Ligand Expression in Bone Marrow Is Associated With Susceptibility to the Development of an Asthmatic Phenotype L. A. Rosenthal1, R. J. Szakaly1, R. L. Sorkness1,2, R. F. Lemanske, Jr.1; 1University of Wisconsin Medical School, Madison, WI, 2School of Pharmacy, University of Wisconsin-Madison, Madison, WI. RATIONALE: After Sendai virus-induced bronchiolitis as weanlings, BN, unlike F344, rats develop a postbronchiolitis asthma-like phenotype, which is prevented by supplemental IFN- therapy. Innate mechanisms of Sendai virus-induced IFN- production are markedly deficient in BN, compared with F344, weanlings. Incubation with GM-CSF or Flt-3 ligand, which are important for the development of the innate immune system, markedly enhanced Sendai virus-induced IFN- production by splenocytes from BN, but not F344, weanlings. Because supplementation with GM-CSF or Flt-3 ligand counteracted this deficient weanling BN innate response, we hypothesized that GM-CSF and Flt-3 ligand expression is deficient in BN weanlings. METHODS: RNA was isolated from bone marrow and lungs of BN and F344 weanlings. Real-time quantitative RT-PCR analysis was used to determine GM-CSF and Flt-3 ligand mRNA levels, which were normalized to -actin mRNA levels. RESULTS: Bone marrow cells from BN weanlings expressed significantly lower levels of GM-CSF (2.7-fold lower; p = 0.004) and Flt-3 ligand (1.3-fold lower; p = 0.037) mRNA than did those from F344 weanlings. There were no strain differences in cytokine mRNA levels in the lungs, indicating a tissue-specific deficiency in BN weanlings. CONCLUSIONS: These results demonstrate deficient GM-CSF and Flt3 ligand mRNA expression in the primary site of hematopoiesis, the bone marrow, in BN weanlings. Reduced expression of GM-CSF and Flt-3 ligand could contribute to the deficient NK cell and accessory cell function in response to Sendai virus in BN weanlings. Therefore, attenuated innate anti-viral responses in BN weanlings may be associated with deficiencies in the development of their innate immune system. Funding: NIH grant AI50500
464
Immune Response in Asthma: Down Modulation by Schistosoma mansoni Infection M. I. Araujo1,2, J. P. Figueiredo1, R. R. Oliveira1, M. C. F. Almeida1, L. M. Alcantara3, B. Hoppe1, A. Schriefer4, E. M. Carvalho1, M. Medeiros, Jr.1; 1Servico de Imunologia, Universidade Federal da Bahia, Salvador,
465
J ALLERGY CLIN IMMUNOL FEBRUARY 2005
Bahia, BRAZIL, 2Instituto de Investigação em Imunologia/CNPq, Brazil, Salvador, Bahia, BRAZIL, 3Faculdade de Farmacia, Universidade Federal da Bahia, Salvador, Bahia, BRAZIL, 4Faculdade de Medicina, Universidade Federal da Bahia, Salvador, Bahia, BRAZIL. RATIONALE: Allergic diseases are highly prevalent in many areas of the world and lead to production of IL-4, IL-5 and IL-13, which are involved in the pathogenesis of these diseases. IL-10, capable of inhibiting the release of histamine and other mediators by mast cells, is highly produced by PBMC of individuals infected with Schistosoma mansoni, and possibly inhibits the reactivity of skin tests observed in schistosomiasis. In this study we evaluated the D. pteronyssinus-induced immune response of asthmatic individuals infected or not with S. mansoni. The role of IL-10 in modulating the immune response through the addition of the mAb anti-IL-10 or rhIL-10 to PBMC culture stimulated with Der p 1 was also evaluated. METHODS: Thirty-three patients living in a polyhelminthic endemic area, Conde, Brazil were selected for the study and ten asthmatic individuals free of parasites, living in a non-endemic area were used as controls. RESULTS: Asthmatic patients from the polyhelminthic endemic area produced low levels of IL-5 in cultures stimulated with Der p1 antigen, while those from the non-endemic area produced high levels of this cytokine. In contrast, Der p Ag-driven IL-10 production was higher in patients from the endemic area than in the control group. Addition of rhIL-10 to the culture of the controls stimulated with Der p lead to a down regulation of IL-5 production. CONCLUSION: It was demonstrated that IL-10 is produced predominantly by monocytes and CD8+T cell. Production of regulatory cytokine, such as IL-10, could represent important mechanisms of down regulation of the inflammatory response in allergies. Funding: NIH Grant # D43 TW06216; Forgaty EID # D43TW00919; FAPESB Generation of the Aspirin-Exacerbated Respiratory Disease (AERD) Phenotype by Th1/Th2 Cytokines L. Borish1, K. Hise1, J. Negri1, R. Caughey2, J. W. Steinke1; 1Beirne Carter Center for Immunology Research, University Of Virginia Health Systems, Charlottesville, VA, 2Otolaryngology, University Of Virginia Health Systems, Charlottesville, VA. RATIONALE: AERD is characterized by: 1) excessive cysteinyl leukotriene (CysLT) production, 2) increased CysLT responsiveness reflecting markedly increased expression of CysLT receptors (CysLTR), and 3) anaphylactoid eosinophil/mast cell activation secondary to reduced PgE2. We investigated the hypothesis that these changes could reflect the activities of Th1/Th2 cytokines and in particular IL-4. METHODS: Enriched populations of eosinophils, monocytes, and B and T cells were obtained by affinity purification. Fibroblasts and epithelial cells were grown from nasal polyp tissue. CysLTR1 and 2 as well as cox1/cox-2 gene expression were quantified by real time PCR. RESULTS: CysLTR 1 and 2 were expressed on most eosinophils and this was not influenced by IL-4, IL-13, or IFN-. In contrast, few mononuclear cells constitutively expressed these receptors. CysLT1R mRNA was significantly induced by IL-4 on T (2.0-fold) and B lymphocytes (2.79fold) and monocytes (2.01-fold). In contrast, no significant changes were observed with IL-13 or IFN-. CysLT2R were markedly induced on T (24.3-fold) and B cells (12.8-fold) by IL-4 and on all 3 mononuclear cells by IFN-. Finally, IL-4, but not IL-13 or IFN-, significantly inhibited monocyte and eosinophil cox-2 mRNA expression while having no influence on cox-1. No modulation of cox-1/cox-2 was observed in epithelial cells or fibroblasts. CONCLUSIONS: The reported upregulation of CysLTR1 and 2 in AERD reflects its mixed Th1/Th2 milieu and influences of IL-4 and IFN. Inhibition of cox-2 on monocytes and eosinophils in combination with their upregulated expression of LTC4 synthase may explain enhanced CysLT synthesis in this disorder and contribute to the sensitivity towards aspirin. Funding: NIH, Merck Research Labs
466
Association Between Second-Hand Smoke Exposure (SSE) and Enhanced Peripheral Blood Mononuclear Cell (PMNC) IL-13 Production During Infancy D. Gentile, A. Patel, D. Skoner; Department of Pediatrics, Allegheny General Hospital, Pittsburgh, PA. RATIONALE: IL-13 production has been documented in children and adults with asthma and atopy. SSE is recognized as a risk factor for the development of childhood asthma. The purpose of this pilot study was to determine whether there is an association between SSE and PMNC IL-13 production during infancy. METHODS: SSE was assessed by questionnaire, and blood was obtained at 2 weeks and 5 months of age in 51 healthy infants. PMNC were cultured and stimulated with PHA, and supernatants were assayed for IL-13 by EIA. RESULTS: Twenty-three infants had no history of SSE, and 28 infants had a history of SSE. IL-13 production was significantly increased in infants with SSE as compared to those without SSE at both ages (p<0.05). In those with and without SSE, the mean (± SEM) levels of IL-13 production (ng/ml) were 1.4 + 0.4 and 8.0 + 0.3 at 2 weeks of age, and 1.8 + 0.5 and 8.8 + 0.2 at 5 months of age, respectively. CONCLUSIONS: These results demonstrate an association between SSE and enhanced PMNC IL-13 production during infancy. Future studies need to expand upon these sample sizes and explore whether enhanced PMNC IL-13 production is the mechanism by which SSE predisposes to the development of asthma/atopy. Funding: NIH
463
SUNDAY
Deficient GM-CSF and Flt-3 Ligand Expression in Bone Marrow Is Associated With Susceptibility to the Development of an Asthmatic Phenotype L. A. Rosenthal1, R. J. Szakaly1, R. L. Sorkness1,2, R. F. Lemanske, Jr.1; 1University of Wisconsin Medical School, Madison, WI, 2School of Pharmacy, University of Wisconsin-Madison, Madison, WI. RATIONALE: After Sendai virus-induced bronchiolitis as weanlings, BN, unlike F344, rats develop a postbronchiolitis asthma-like phenotype, which is prevented by supplemental IFN- therapy. Innate mechanisms of Sendai virus-induced IFN- production are markedly deficient in BN, compared with F344, weanlings. Incubation with GM-CSF or Flt-3 ligand, which are important for the development of the innate immune system, markedly enhanced Sendai virus-induced IFN- production by splenocytes from BN, but not F344, weanlings. Because supplementation with GM-CSF or Flt-3 ligand counteracted this deficient weanling BN innate response, we hypothesized that GM-CSF and Flt-3 ligand expression is deficient in BN weanlings. METHODS: RNA was isolated from bone marrow and lungs of BN and F344 weanlings. Real-time quantitative RT-PCR analysis was used to determine GM-CSF and Flt-3 ligand mRNA levels, which were normalized to -actin mRNA levels. RESULTS: Bone marrow cells from BN weanlings expressed significantly lower levels of GM-CSF (2.7-fold lower; p = 0.004) and Flt-3 ligand (1.3-fold lower; p = 0.037) mRNA than did those from F344 weanlings. There were no strain differences in cytokine mRNA levels in the lungs, indicating a tissue-specific deficiency in BN weanlings. CONCLUSIONS: These results demonstrate deficient GM-CSF and Flt3 ligand mRNA expression in the primary site of hematopoiesis, the bone marrow, in BN weanlings. Reduced expression of GM-CSF and Flt-3 ligand could contribute to the deficient NK cell and accessory cell function in response to Sendai virus in BN weanlings. Therefore, attenuated innate anti-viral responses in BN weanlings may be associated with deficiencies in the development of their innate immune system. Funding: NIH grant AI50500
464
Immune Response in Asthma: Down Modulation by Schistosoma mansoni Infection M. I. Araujo1,2, J. P. Figueiredo1, R. R. Oliveira1, M. C. F. Almeida1, L. M. Alcantara3, B. Hoppe1, A. Schriefer4, E. M. Carvalho1, M. Medeiros, Jr.1; 1Servico de Imunologia, Universidade Federal da Bahia, Salvador,
465
J ALLERGY CLIN IMMUNOL FEBRUARY 2005
Bahia, BRAZIL, 2Instituto de Investigação em Imunologia/CNPq, Brazil, Salvador, Bahia, BRAZIL, 3Faculdade de Farmacia, Universidade Federal da Bahia, Salvador, Bahia, BRAZIL, 4Faculdade de Medicina, Universidade Federal da Bahia, Salvador, Bahia, BRAZIL. RATIONALE: Allergic diseases are highly prevalent in many areas of the world and lead to production of IL-4, IL-5 and IL-13, which are involved in the pathogenesis of these diseases. IL-10, capable of inhibiting the release of histamine and other mediators by mast cells, is highly produced by PBMC of individuals infected with Schistosoma mansoni, and possibly inhibits the reactivity of skin tests observed in schistosomiasis. In this study we evaluated the D. pteronyssinus-induced immune response of asthmatic individuals infected or not with S. mansoni. The role of IL-10 in modulating the immune response through the addition of the mAb anti-IL-10 or rhIL-10 to PBMC culture stimulated with Der p 1 was also evaluated. METHODS: Thirty-three patients living in a polyhelminthic endemic area, Conde, Brazil were selected for the study and ten asthmatic individuals free of parasites, living in a non-endemic area were used as controls. RESULTS: Asthmatic patients from the polyhelminthic endemic area produced low levels of IL-5 in cultures stimulated with Der p1 antigen, while those from the non-endemic area produced high levels of this cytokine. In contrast, Der p Ag-driven IL-10 production was higher in patients from the endemic area than in the control group. Addition of rhIL-10 to the culture of the controls stimulated with Der p lead to a down regulation of IL-5 production. CONCLUSION: It was demonstrated that IL-10 is produced predominantly by monocytes and CD8+T cell. Production of regulatory cytokine, such as IL-10, could represent important mechanisms of down regulation of the inflammatory response in allergies. Funding: NIH Grant # D43 TW06216; Forgaty EID # D43TW00919; FAPESB Generation of the Aspirin-Exacerbated Respiratory Disease (AERD) Phenotype by Th1/Th2 Cytokines L. Borish1, K. Hise1, J. Negri1, R. Caughey2, J. W. Steinke1; 1Beirne Carter Center for Immunology Research, University Of Virginia Health Systems, Charlottesville, VA, 2Otolaryngology, University Of Virginia Health Systems, Charlottesville, VA. RATIONALE: AERD is characterized by: 1) excessive cysteinyl leukotriene (CysLT) production, 2) increased CysLT responsiveness reflecting markedly increased expression of CysLT receptors (CysLTR), and 3) anaphylactoid eosinophil/mast cell activation secondary to reduced PgE2. We investigated the hypothesis that these changes could reflect the activities of Th1/Th2 cytokines and in particular IL-4. METHODS: Enriched populations of eosinophils, monocytes, and B and T cells were obtained by affinity purification. Fibroblasts and epithelial cells were grown from nasal polyp tissue. CysLTR1 and 2 as well as cox1/cox-2 gene expression were quantified by real time PCR. RESULTS: CysLTR 1 and 2 were expressed on most eosinophils and this was not influenced by IL-4, IL-13, or IFN-. In contrast, few mononuclear cells constitutively expressed these receptors. CysLT1R mRNA was significantly induced by IL-4 on T (2.0-fold) and B lymphocytes (2.79fold) and monocytes (2.01-fold). In contrast, no significant changes were observed with IL-13 or IFN-. CysLT2R were markedly induced on T (24.3-fold) and B cells (12.8-fold) by IL-4 and on all 3 mononuclear cells by IFN-. Finally, IL-4, but not IL-13 or IFN-, significantly inhibited monocyte and eosinophil cox-2 mRNA expression while having no influence on cox-1. No modulation of cox-1/cox-2 was observed in epithelial cells or fibroblasts. CONCLUSIONS: The reported upregulation of CysLTR1 and 2 in AERD reflects its mixed Th1/Th2 milieu and influences of IL-4 and IFN. Inhibition of cox-2 on monocytes and eosinophils in combination with their upregulated expression of LTC4 synthase may explain enhanced CysLT synthesis in this disorder and contribute to the sensitivity towards aspirin. Funding: NIH, Merck Research Labs
466