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CCR5 Usage by CCL5 Induces a Selective Leukocyte Recruitment in Human Skin Xenografts in Vivo
Abstracts S267
J ALLERGY CLIN IMMUNOL VOLUME 117, NUMBER 2
CCR5 Usage by CCL5 Induces a Selective Leukocyte Recruitment in Human Skin Xenografts in Vivo A. Tsicopoulos1, P. de Nadai1, C. Chenivesse1, H. Porte2, H. Vorng1, A. F. Walls3, B. Wallaert1, A. Tonnel1, H. Zerwes4; 1Institut Pasteur de Lille, U416 INSERM, Lille cedex, FRANCE, 2Service de Chirurgie, CHRU de Lille, Lille cedex, FRANCE, 3Immunopharmacology group, Southampton general hospital, Southampton, UNITED KINGDOM, 4Novartis, basel, SWITZERLAND. RATIONALE: CCR5 is one of the major chemokine receptors with potential therapeutical applications in humans. However, the redundancy of chemokines and their receptors, and the species specificity of chemokine receptor antagonists question the validity of this approach. METHODS: we used a humanised SCID mouse model grafted with human skin and autologous leukocytes, and evaluated the effect of a blocking antibody against human CCR5, on CCL5-induced cutaneous leukocyte recruitment in vivo. RESULTS: At baseline, CCL5 induced a significant recruitment of T cells mainly of the memory phenotype, of monocytes/macrophages and eosinophils,and of IFN-g+ but not IL-4+ and IL-5+ cells. In vivo, antiCCR5 antibody was able to almost completely inhibit the recruitment of monocytes/macrophages and Th-1 type cells, to partially inhibit the attraction of memory T cells, but had no effects on eosinophil infiltration, although besides CCR5, all these cell types express other CCL5 binding chemokine receptors. CONCLUSIONS: These results indicate that the in vivo environment regulates target cell specificity of CCL5 leading to differential cell recruitment suggesting that antagonizing CCR5 receptor may be of therapeutic value in diseases where CCL5/CCR5, monocytes and Th1 type cells play a predominant role. Funding: INSERM
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Immunologic Changes in Human Albumin Exposed to Toluene Diisocyanate (TDI) Vapor: Differences Compared with Liquid TDI Exposure and Association with IgE/IgG Binding and TDI Asthma Y. Ye1, S. Kim2, S. Lee, III3, H. Park1, C. A. Redlich4, A. V. Wisnewski4; 1Dept. of Allergy & Rheumatology, Ajou University School of Medicine, Suwon, REPUBLIC OF KOREA, 2Dept. of Internal Medicine, Yonsei University Wonju College of Medicine, Wonju, REPUBLIC OF KOREA, 3Donga University School of Medicine, Busan, REPUBLIC OF KOREA, 4Yale University School of Medicine, New Haven, CT. RATIONALE: The extreme reactivity of TDI has hampered serologic studies on TDI asthma due to uncertainty regarding the “antigenic” form of this chemical. Determine the influence of vapor vs. liquid exposure on the generation of TDI-albumin conjugates and the outcome of serology assays for TDI-specific IgE and IgG.
1033
METHODS: TDI-albumin conjugates were prepared by vapor TDI exposure (vapor TDI-albumin) and liquid phase TDI exposure methods (liquid TDI-albumin), and two different mixtures of 2,4- and 2,6-TDI isomers were tested. The TDI-albumin conjugates were used as antigens in ELISA assays to measure specific IgE and IgG antibodies in sera from 66 patients with TDI-induced asthma, 167 asymptomatic exposed subjects, 64 patients with allergic asthma, and 123 unexposed healthy controls. RESULTS: Among TDI asthmatics, the prevalence of specific IgE and IgG that bound vapor TDI-albumin conjugates was higher than that which bound liquid TDI-albumin conjugates, especially those prepared with an 80:20 mixture of 2,4:2,6-TDI. Furthermore, the prevalence of IgE and IgG that bound vapor TDI-albumin conjugates was significantly higher among TDI asthmatics than the control groups (44% vs. < 5% and 31% vs. < 10% for IgE and IgG respectively). CONCLUSIONS: Vapor TDI-albumin conjugates differ from those prepared by previously published methods and may serve as better diagnostic markers for identifying asthmatic patients among TDI-exposed workers, and as reagents for studies on immunopathogenesis. Funding: Koreea Health 21 R&D project, Ministry of Health & Welfare Prediction of Specific Airway Responsiveness from Skin Sensitivity to Allergen and Airway Hyperresponsiveness to Methacholine in Baker’s Asthma S. Quirce, M. Fernández-Nieto, C. Escudero, J. Cuesta, M. de las Heras, J. Sastre; Allergy Department, Fundación Jiménez Díaz, Madrid, SPAIN. RATIONALE: Relationships between immunological reactivity, nonspecific airway hyperresponsiveness and bronchial responsiveness to allergen have been scarcely investigated in occupational asthma. METHODS: We assessed the above relationships in 24 subjects with baker’s asthma. The skin endpoint titration to bakery allergens as a measure of immunological reactivity, together with the methacholine PC20 and allergen PC20 during the early asthmatic response were determined. RESULTS: All patients had positive skin tests to some bakery allergens (wheat flour, rye flour, soybean flour, fungal enzymes and egg white proteins). Methacholine inhalation tests revealed bronchial hyperresponsiveness in all of them. Specific inhalation challenge (SIC) tests with aqueous allergen extracts were performed with wheat flour (n=12), rye flour (n=2), soybean flour (n=6), soybean trypsin inhibitor (n=2), alpha-amylase (n=4), hemicellulase (n=5), glucoamylase (n=3), ovalbumin (n=2), ovomucoid (n=1) and lysozyme (n=5) in sensitized workers. A positive asthmatic response was observed in 84% of the inhalation challenges. SIC elicited isolated early asthmatic responses in 63% cases, dual responses in 32% and isolated late responses in 5%. Multiple linear regression analysis showed allergen PC20 as a function of skin sensitivity to allergen and methacholine PC20, yielding the following highly significant regression formula: log allergen PC20= 0.18 + 0.99 log (skin sensitivity) + 0.343 log (methacholine PC20) (r=0.89, p<0.001). This formula predicted allergen PC20 to within one double concentration in 67%, to within two double concentrations in 85% and within 3 double concentrations in 97%. CONCLUSIONS: Airway responsiveness to bakery allergens can be satisfactorily predicted by assessing skin sensitivity to allergen and bronchial responsiveness to methacholine.
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comprised primarily of VSMC. In CX3CR1-/- mice, there was no detectable monocyte invasion into the intima at d5 (p=0.006). At d28, the intima area was decreased in CX3CR1-/- mice by 80% (p=0.003), with a concomitant decrease in the numbers of VSMC in the intima (87.1%, p=0.05). The percentage of actively proliferating cells in injured vessels was also decreased in CX3CR1-/- mice (12% vs. 27% in WT, p=0.03). CONCLUSIONS: CX3CR1 plays a critical role in vascular inflammation following arterial injury, and one of the primary mechanisms involves monocyte recruitment to and/or retention within the injured vessel intima.
J ALLERGY CLIN IMMUNOL VOLUME 117, NUMBER 2
CCR5 Usage by CCL5 Induces a Selective Leukocyte Recruitment in Human Skin Xenografts in Vivo A. Tsicopoulos1, P. de Nadai1, C. Chenivesse1, H. Porte2, H. Vorng1, A. F. Walls3, B. Wallaert1, A. Tonnel1, H. Zerwes4; 1Institut Pasteur de Lille, U416 INSERM, Lille cedex, FRANCE, 2Service de Chirurgie, CHRU de Lille, Lille cedex, FRANCE, 3Immunopharmacology group, Southampton general hospital, Southampton, UNITED KINGDOM, 4Novartis, basel, SWITZERLAND. RATIONALE: CCR5 is one of the major chemokine receptors with potential therapeutical applications in humans. However, the redundancy of chemokines and their receptors, and the species specificity of chemokine receptor antagonists question the validity of this approach. METHODS: we used a humanised SCID mouse model grafted with human skin and autologous leukocytes, and evaluated the effect of a blocking antibody against human CCR5, on CCL5-induced cutaneous leukocyte recruitment in vivo. RESULTS: At baseline, CCL5 induced a significant recruitment of T cells mainly of the memory phenotype, of monocytes/macrophages and eosinophils,and of IFN-g+ but not IL-4+ and IL-5+ cells. In vivo, antiCCR5 antibody was able to almost completely inhibit the recruitment of monocytes/macrophages and Th-1 type cells, to partially inhibit the attraction of memory T cells, but had no effects on eosinophil infiltration, although besides CCR5, all these cell types express other CCL5 binding chemokine receptors. CONCLUSIONS: These results indicate that the in vivo environment regulates target cell specificity of CCL5 leading to differential cell recruitment suggesting that antagonizing CCR5 receptor may be of therapeutic value in diseases where CCL5/CCR5, monocytes and Th1 type cells play a predominant role. Funding: INSERM
1032
Immunologic Changes in Human Albumin Exposed to Toluene Diisocyanate (TDI) Vapor: Differences Compared with Liquid TDI Exposure and Association with IgE/IgG Binding and TDI Asthma Y. Ye1, S. Kim2, S. Lee, III3, H. Park1, C. A. Redlich4, A. V. Wisnewski4; 1Dept. of Allergy & Rheumatology, Ajou University School of Medicine, Suwon, REPUBLIC OF KOREA, 2Dept. of Internal Medicine, Yonsei University Wonju College of Medicine, Wonju, REPUBLIC OF KOREA, 3Donga University School of Medicine, Busan, REPUBLIC OF KOREA, 4Yale University School of Medicine, New Haven, CT. RATIONALE: The extreme reactivity of TDI has hampered serologic studies on TDI asthma due to uncertainty regarding the “antigenic” form of this chemical. Determine the influence of vapor vs. liquid exposure on the generation of TDI-albumin conjugates and the outcome of serology assays for TDI-specific IgE and IgG.
1033
METHODS: TDI-albumin conjugates were prepared by vapor TDI exposure (vapor TDI-albumin) and liquid phase TDI exposure methods (liquid TDI-albumin), and two different mixtures of 2,4- and 2,6-TDI isomers were tested. The TDI-albumin conjugates were used as antigens in ELISA assays to measure specific IgE and IgG antibodies in sera from 66 patients with TDI-induced asthma, 167 asymptomatic exposed subjects, 64 patients with allergic asthma, and 123 unexposed healthy controls. RESULTS: Among TDI asthmatics, the prevalence of specific IgE and IgG that bound vapor TDI-albumin conjugates was higher than that which bound liquid TDI-albumin conjugates, especially those prepared with an 80:20 mixture of 2,4:2,6-TDI. Furthermore, the prevalence of IgE and IgG that bound vapor TDI-albumin conjugates was significantly higher among TDI asthmatics than the control groups (44% vs. < 5% and 31% vs. < 10% for IgE and IgG respectively). CONCLUSIONS: Vapor TDI-albumin conjugates differ from those prepared by previously published methods and may serve as better diagnostic markers for identifying asthmatic patients among TDI-exposed workers, and as reagents for studies on immunopathogenesis. Funding: Koreea Health 21 R&D project, Ministry of Health & Welfare Prediction of Specific Airway Responsiveness from Skin Sensitivity to Allergen and Airway Hyperresponsiveness to Methacholine in Baker’s Asthma S. Quirce, M. Fernández-Nieto, C. Escudero, J. Cuesta, M. de las Heras, J. Sastre; Allergy Department, Fundación Jiménez Díaz, Madrid, SPAIN. RATIONALE: Relationships between immunological reactivity, nonspecific airway hyperresponsiveness and bronchial responsiveness to allergen have been scarcely investigated in occupational asthma. METHODS: We assessed the above relationships in 24 subjects with baker’s asthma. The skin endpoint titration to bakery allergens as a measure of immunological reactivity, together with the methacholine PC20 and allergen PC20 during the early asthmatic response were determined. RESULTS: All patients had positive skin tests to some bakery allergens (wheat flour, rye flour, soybean flour, fungal enzymes and egg white proteins). Methacholine inhalation tests revealed bronchial hyperresponsiveness in all of them. Specific inhalation challenge (SIC) tests with aqueous allergen extracts were performed with wheat flour (n=12), rye flour (n=2), soybean flour (n=6), soybean trypsin inhibitor (n=2), alpha-amylase (n=4), hemicellulase (n=5), glucoamylase (n=3), ovalbumin (n=2), ovomucoid (n=1) and lysozyme (n=5) in sensitized workers. A positive asthmatic response was observed in 84% of the inhalation challenges. SIC elicited isolated early asthmatic responses in 63% cases, dual responses in 32% and isolated late responses in 5%. Multiple linear regression analysis showed allergen PC20 as a function of skin sensitivity to allergen and methacholine PC20, yielding the following highly significant regression formula: log allergen PC20= 0.18 + 0.99 log (skin sensitivity) + 0.343 log (methacholine PC20) (r=0.89, p<0.001). This formula predicted allergen PC20 to within one double concentration in 67%, to within two double concentrations in 85% and within 3 double concentrations in 97%. CONCLUSIONS: Airway responsiveness to bakery allergens can be satisfactorily predicted by assessing skin sensitivity to allergen and bronchial responsiveness to methacholine.
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comprised primarily of VSMC. In CX3CR1-/- mice, there was no detectable monocyte invasion into the intima at d5 (p=0.006). At d28, the intima area was decreased in CX3CR1-/- mice by 80% (p=0.003), with a concomitant decrease in the numbers of VSMC in the intima (87.1%, p=0.05). The percentage of actively proliferating cells in injured vessels was also decreased in CX3CR1-/- mice (12% vs. 27% in WT, p=0.03). CONCLUSIONS: CX3CR1 plays a critical role in vascular inflammation following arterial injury, and one of the primary mechanisms involves monocyte recruitment to and/or retention within the injured vessel intima.