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Analysis of Circulating CD25hiCD4+ T Cells in Atopic Dermatitis Reveals Age-Related Changes in Early Life
S32 Abstracts
SATURDAY
J ALLERGY CLIN IMMUNOL FEBRUARY 2008
123
The Tight Junction Protein, Claudin-1 is Dysregulated in Atopic Dermatitis A. De Benedetto1, L. R. Latchney1, L. Y. McGirt2, S. Vidyasagar1, C. Cheadle2, K. C. Barnes2, L. A. Beck1; 1University of Rochester, Rochester, NY, 2Johns Hopkins University School of Medicine, Baltimore, MD. RATIONALE: Tight junctions (TJ) provide mechanical support and are a barrier to the diffusion of fluid/electrolytes, macromolecules and pathogens. Murine models suggest that TJs, more specifically Claudins, play a central role in cutaneous barrier. We utilized expression profiling to evaluate the expression of these proteins in epithelium from atopic dermatitis (AD) and nonatopic, healthy controls (NA). METHODS: The suction-blister approach was employed to obtain nonlesional epithelium from subjects with extrinsic AD (n 5 5) and NA (n 5 5). Expression profiles were generated using Illumina’s Sentrix HumanRef-8 Expression BeadChips, analyzed using Gene Set Matrix Analysis. We also evaluated the modulation of Claudin-1 in cultured primary keratinocytes (KC) by Western blot. RESULTS: Genes related to intercellular junctions were dysregulated in AD vs NA. We noted downregulation of the TJ proteins, Claudin-1 and Claudin-23 and Desmoglein-1 in AD. Staining of nonlesional AD skin (n 5 11) confirmed this reduced expression of Claudin-1, while in lesional AD skin (n 5 11) it was upregulated compared to NA (n 5 12). Claudin1 protein expression was induced by differentiation of KC and stimulation with PGN, Der p1, IL-4 and -13. CONCLUSIONS: Our studies suggest that an effective barrier in pseudostratified squamous epithelium may be determined by the relative expression of proteins important for formation of the cornified envelope (e.g. Filaggrin) and TJs. The barrier defect that results from dysregulation of these proteins probably results in greater penetration of allergens, irritants and microbes. We believe the compensatory increase in Claudin1 observed in lesional AD skin may be due to the local actions of Th2 cytokines, allergen exposure or staphylococcal colonization. Funding: National Eczema Association; Atopic Dermatitis Vaccinia Network (NOI AI40029)
Safety Of Omalizumab Treatment In Children (> 4 Years) With High Serum IgE and Severe Atopic Dermatitis And Food Allergy S. R. Iyengar1, E. G. Hoyte2, A. Loza2, S. Bonaccorso3, D. T. Umetsu1, K. C. Nadeau2; 1Division of Immunology, Children’s Hospital, Boston, MA, 2Department of Pediatrics, Stanford University, Stanford, CA, 3 Department of Pediatrics, Stanford University, Stanford, CA. RATIONALE: Omalizumab is approved for treatment of moderate to severe asthma, but its safety in young children (<12 years) with serum IgE >700 IU/mL has not been established. METHODS: We examined the safety of omalizumab in a double blind, placebo controlled Phase I study in subjects aged 4-22 yrs (mean 5 11.6 yrs) with severe atopic dermatitis (AD), food allergy and elevated serum IgE (range 5 105-2800; mean 5 1,453 IU/mL). The study was FDAsanctioned, and subjects received omalizumab (n 5 4) or placebo (n 5 4) every 2-4 weeks subcutaneously over 24 weeks. Subjects received omalizumab at doses that accommodated high serum IgE levels, using a regimen extrapolated from omalizumab package insert dosages. Blood counts, chemistries, and IgE levels were checked monthly. Adverse events (AE) and clinical status were monitored per NIH guidelines. RESULTS: In the treatment group, there was one serious AE in which a subject was hospitalized for cellulitis, and 5 non-serious AEs, including an asthma flare, skin infection, leg pain, conjunctivitis, and epistaxis. In the control group, there were 6 non-serious AEs; including an URI, conjunctivitis and AD, asthma, and eosinophilic esophagitis flares. None of the AEs were judged to be related to the study medication. No clinically significant abnormal laboratory values were observed. CONCLUSIONS: There was no significant difference in adverse events or clinical laboratory studies in the omalizumab-treated compared to placebo-treated subjects using this dosing regimen in young children with high serum IgE, AD, and food allergy. Therefore, omalizumab appears to be safe in young children with elevated serum IgE with a higher dosing regimen. Funding: Oxnard Foundation
Analysis of Circulating CD25hiCD41 T Cells in Atopic Dermatitis Reveals Age-Related Changes in Early Life T. W. Song, A. J. Reefer, M. Moloney, M. D. Solga, S. M. Satinover, D. D. Murphy, P. W. Heymann, J. A. Woodfolk; University of Virginia Health System, Charlottesville, VA. RATIONALE: Circulating CD25hiCD41 T lymphocytes which express the chemokine receptor, CCR4, are increased in adults with atopic dermatitis (AD). Little is known about the properties of CD25hi T cells in early life and how they evolve. Our objective was to analyze these cells in children with AD. METHODS: PBMCs were isolated from children (ages 2 to 13 years, n 5 8) who presented with eczematous skin eruption. CD25hi T cells were analyzed by flow cytometry for expression of surface markers and Foxp3. RESULTS: CD25hi T cells were detectable from aged 2; however, CCR4 was preferentially expressed on these cells only after 3 years of age. The frequency of CD25hiCCR41 T cells was highest in children >7 years (up to 8.7% of CD41 T cells) who were highly atopic (total IgE >5,000 IU/ ml). Foxp31 T cells were enriched within the CD25hi subset (range 15% to 92%) as compared with CD25med T cells (range 1% to 9%) and these cells were CD127lo/neg. Notably, the percentage of CD25hi T cells which were Foxp31 increased with age (r 5 0.84, p 5 0.009) and analysis of T cells from healthy age-matched controls suggested that this reflected normal maturation. The frequency of CD25hi T cells expressing the skinhoming addressin, CLA, was highly variable (5% to 46%). Moreover, in contrast to adult patients, the majority of CLA1 T cells were CD25negative, irrespective of age. CONCLUSIONS: Analysis of evolving CD25hiCD41 T cells in children with AD exposes important age-related changes which could be relevant to the capacity of these cells to traffic to skin and function effectively. Funding: NIH/NIAID
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The Role of Asthma in Remission of Atopic Dermatitis A. K. Glasofer, E. Yousef; Thomas Jefferson University Hospital, Philadelphia, PA. RATIONALE: IgE mediated diseases and asthma have long been implicated as playing a major role in atopic dermatitis (AD). AD has been viewed as a first step in the ‘‘Atopic March’’ with the possibility of the development of asthma. However, the role of asthma in the natural history of AD is not clear. The purpose of this study was to investigate the role of asthma in the remission of atopic dermatitis. METHODS: Medical records of children aged 5 years or more, diagnosed with AD were reviewed. ‘‘Remission’’ was defined as 1) Two normal skin exams within a 6-month period without the use of rescue medications, or 2) Follow up within 1 year of last exam documenting no AD flares or need for rescue medication during the interval. Patients were excluded if they had no documented skin exam for one year, and were considered lost to follow up. Patient charts were further analyzed to look for the co-morbidity of asthma. RESULTS: Of the 60 patients who met inclusion criteria, 15 (25%) had remission of their AD (Group A) and 45 (75%) did not have remission of their AD (Group B). Within Group A, 87% (n 5 13) had asthma, while 75% (n 5 34) of Group B had asthma. 78% (n 5 47) of all patients included in the study had asthma. CONCLUSIONS: Asthma does not affect remission of AD, but the ‘‘Atopic March’’ proceeds whether or not there is remission of AD.
SATURDAY
J ALLERGY CLIN IMMUNOL FEBRUARY 2008
123
The Tight Junction Protein, Claudin-1 is Dysregulated in Atopic Dermatitis A. De Benedetto1, L. R. Latchney1, L. Y. McGirt2, S. Vidyasagar1, C. Cheadle2, K. C. Barnes2, L. A. Beck1; 1University of Rochester, Rochester, NY, 2Johns Hopkins University School of Medicine, Baltimore, MD. RATIONALE: Tight junctions (TJ) provide mechanical support and are a barrier to the diffusion of fluid/electrolytes, macromolecules and pathogens. Murine models suggest that TJs, more specifically Claudins, play a central role in cutaneous barrier. We utilized expression profiling to evaluate the expression of these proteins in epithelium from atopic dermatitis (AD) and nonatopic, healthy controls (NA). METHODS: The suction-blister approach was employed to obtain nonlesional epithelium from subjects with extrinsic AD (n 5 5) and NA (n 5 5). Expression profiles were generated using Illumina’s Sentrix HumanRef-8 Expression BeadChips, analyzed using Gene Set Matrix Analysis. We also evaluated the modulation of Claudin-1 in cultured primary keratinocytes (KC) by Western blot. RESULTS: Genes related to intercellular junctions were dysregulated in AD vs NA. We noted downregulation of the TJ proteins, Claudin-1 and Claudin-23 and Desmoglein-1 in AD. Staining of nonlesional AD skin (n 5 11) confirmed this reduced expression of Claudin-1, while in lesional AD skin (n 5 11) it was upregulated compared to NA (n 5 12). Claudin1 protein expression was induced by differentiation of KC and stimulation with PGN, Der p1, IL-4 and -13. CONCLUSIONS: Our studies suggest that an effective barrier in pseudostratified squamous epithelium may be determined by the relative expression of proteins important for formation of the cornified envelope (e.g. Filaggrin) and TJs. The barrier defect that results from dysregulation of these proteins probably results in greater penetration of allergens, irritants and microbes. We believe the compensatory increase in Claudin1 observed in lesional AD skin may be due to the local actions of Th2 cytokines, allergen exposure or staphylococcal colonization. Funding: National Eczema Association; Atopic Dermatitis Vaccinia Network (NOI AI40029)
Safety Of Omalizumab Treatment In Children (> 4 Years) With High Serum IgE and Severe Atopic Dermatitis And Food Allergy S. R. Iyengar1, E. G. Hoyte2, A. Loza2, S. Bonaccorso3, D. T. Umetsu1, K. C. Nadeau2; 1Division of Immunology, Children’s Hospital, Boston, MA, 2Department of Pediatrics, Stanford University, Stanford, CA, 3 Department of Pediatrics, Stanford University, Stanford, CA. RATIONALE: Omalizumab is approved for treatment of moderate to severe asthma, but its safety in young children (<12 years) with serum IgE >700 IU/mL has not been established. METHODS: We examined the safety of omalizumab in a double blind, placebo controlled Phase I study in subjects aged 4-22 yrs (mean 5 11.6 yrs) with severe atopic dermatitis (AD), food allergy and elevated serum IgE (range 5 105-2800; mean 5 1,453 IU/mL). The study was FDAsanctioned, and subjects received omalizumab (n 5 4) or placebo (n 5 4) every 2-4 weeks subcutaneously over 24 weeks. Subjects received omalizumab at doses that accommodated high serum IgE levels, using a regimen extrapolated from omalizumab package insert dosages. Blood counts, chemistries, and IgE levels were checked monthly. Adverse events (AE) and clinical status were monitored per NIH guidelines. RESULTS: In the treatment group, there was one serious AE in which a subject was hospitalized for cellulitis, and 5 non-serious AEs, including an asthma flare, skin infection, leg pain, conjunctivitis, and epistaxis. In the control group, there were 6 non-serious AEs; including an URI, conjunctivitis and AD, asthma, and eosinophilic esophagitis flares. None of the AEs were judged to be related to the study medication. No clinically significant abnormal laboratory values were observed. CONCLUSIONS: There was no significant difference in adverse events or clinical laboratory studies in the omalizumab-treated compared to placebo-treated subjects using this dosing regimen in young children with high serum IgE, AD, and food allergy. Therefore, omalizumab appears to be safe in young children with elevated serum IgE with a higher dosing regimen. Funding: Oxnard Foundation
Analysis of Circulating CD25hiCD41 T Cells in Atopic Dermatitis Reveals Age-Related Changes in Early Life T. W. Song, A. J. Reefer, M. Moloney, M. D. Solga, S. M. Satinover, D. D. Murphy, P. W. Heymann, J. A. Woodfolk; University of Virginia Health System, Charlottesville, VA. RATIONALE: Circulating CD25hiCD41 T lymphocytes which express the chemokine receptor, CCR4, are increased in adults with atopic dermatitis (AD). Little is known about the properties of CD25hi T cells in early life and how they evolve. Our objective was to analyze these cells in children with AD. METHODS: PBMCs were isolated from children (ages 2 to 13 years, n 5 8) who presented with eczematous skin eruption. CD25hi T cells were analyzed by flow cytometry for expression of surface markers and Foxp3. RESULTS: CD25hi T cells were detectable from aged 2; however, CCR4 was preferentially expressed on these cells only after 3 years of age. The frequency of CD25hiCCR41 T cells was highest in children >7 years (up to 8.7% of CD41 T cells) who were highly atopic (total IgE >5,000 IU/ ml). Foxp31 T cells were enriched within the CD25hi subset (range 15% to 92%) as compared with CD25med T cells (range 1% to 9%) and these cells were CD127lo/neg. Notably, the percentage of CD25hi T cells which were Foxp31 increased with age (r 5 0.84, p 5 0.009) and analysis of T cells from healthy age-matched controls suggested that this reflected normal maturation. The frequency of CD25hi T cells expressing the skinhoming addressin, CLA, was highly variable (5% to 46%). Moreover, in contrast to adult patients, the majority of CLA1 T cells were CD25negative, irrespective of age. CONCLUSIONS: Analysis of evolving CD25hiCD41 T cells in children with AD exposes important age-related changes which could be relevant to the capacity of these cells to traffic to skin and function effectively. Funding: NIH/NIAID
126
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125
The Role of Asthma in Remission of Atopic Dermatitis A. K. Glasofer, E. Yousef; Thomas Jefferson University Hospital, Philadelphia, PA. RATIONALE: IgE mediated diseases and asthma have long been implicated as playing a major role in atopic dermatitis (AD). AD has been viewed as a first step in the ‘‘Atopic March’’ with the possibility of the development of asthma. However, the role of asthma in the natural history of AD is not clear. The purpose of this study was to investigate the role of asthma in the remission of atopic dermatitis. METHODS: Medical records of children aged 5 years or more, diagnosed with AD were reviewed. ‘‘Remission’’ was defined as 1) Two normal skin exams within a 6-month period without the use of rescue medications, or 2) Follow up within 1 year of last exam documenting no AD flares or need for rescue medication during the interval. Patients were excluded if they had no documented skin exam for one year, and were considered lost to follow up. Patient charts were further analyzed to look for the co-morbidity of asthma. RESULTS: Of the 60 patients who met inclusion criteria, 15 (25%) had remission of their AD (Group A) and 45 (75%) did not have remission of their AD (Group B). Within Group A, 87% (n 5 13) had asthma, while 75% (n 5 34) of Group B had asthma. 78% (n 5 47) of all patients included in the study had asthma. CONCLUSIONS: Asthma does not affect remission of AD, but the ‘‘Atopic March’’ proceeds whether or not there is remission of AD.