Investigation of the Mechanisms of Pressure Ulcers Using a Cyclical Cutaneous Ischemic–reperfusion Injury Model

S164 by flow cytometry on PBMC from RF and also on PBMC from healthy controls (HC). Results. IFN-γ+cells in RF-1 = 38.5 vs. 15 HC (p = 0.0016); TNF-α+cells in RF-1 = 55 vs. 3.2 HC (p = 0.0015); IFN-γ+cells in RF-2 = 32.6 vs. 8.1 HC (p = 0.026); TNF-α+cells in RF-2 = 34.6 vs. 1.5 HC (p = 0.0056); IFN-γ+cells in RF-2 = 32.6 vs. 8.1 HC (p = 0.026); TNF-α+cells in RF-2 = 34.6 vs. 1.5 HC (p = 0.0056); IFN-γ+cells in RF-3 = 24.2 vs. 8.3 HC (p = 0.04); TNF-α+cells in RF-3 = 32.2 vs. 3.1 HC (p = 0.0046). CONCLUSIONS. Peripheral blood mononuclear cells from a Rosacea family have significant increase in the expression of IFN-γ+ and TNF-α+ cells. IFN-γ and TNF-α may have a role in the inflammatory process of rosacea stimulating the release of reactive oxygen species in macrophages and neutrophils causing damage to facial follicles in persons with Rosacea. doi:10.1016/j.clim.2007.03.105

Su.65 Does Quality of Life in Cutaneous Lupus Erythematosus Correlate with Disease Severity? Elizabeth Gaines, Pre-doctoral Research Fellow, University of Pennsylvania, Department of Dermatology, Philadelphia, PA The purpose of this study was to assess the correlation between cutaneous lupus erythematosus (CLE) disease severity, as measured by the CLASI, and quality-of-life (QoL), as measured by the Skindex-29. This was a prospective, longitudinal study from baseline (Day 0) to Day 56, done at a University hospital cutaneous autoimmunity clinic. Eight patients with biopsy-confirmed CLE (4 DLE, 2 SCLE, 2 DLE/ SLE) completed the study. At each visit, patients completed the CLE-modified Skindex-29. Using scales from 0-10, they also assessed their general and skin health, and their pain, itch, and fatigue. The PI evaluated disease severity, with the CLASI, and the patient s global skin health using a scale from 0−10. There was a significant improvement in total Skindex score over time (t=2.36, pb0.05). There was a moderate correlation (r=0.59, p=0.12, n=8) between change in activity (CLASI), and change in skin symptoms (Skindex). Neither change in emotion or function correlated with change in activity. There was no correlation between any Skindex subscale with damage. Skindex scores for patients with SCLE were, on average, lower and changed less than Skindex scores for patients with DLE. While both the CLASI activity score and the modified Skindex total score improved significantly over time, the correlation between the two scores was poor. Different elements of skin health may be captured by each index. Given the different disease courses and likelihood of damage, skin-related QoL may differ across different subtypes of CLE. Preliminary analysis of data from 25 additional CLE patients supports these conclusions. doi:10.1016/j.clim.2007.03.106

Su.66 Association Between the Polymorphism of TGF-β1 Gene Promoter (−509CNT) and Idiopathic Chronic Urticaria Sara Hosseini-Farahabadi, Dr.; Researcher, Bu-Ali Research Institute (BARI), Department of Immunogenetics, Mashhad, Iran, Jalil Tavakkol-Afshari, Vice President of Research;

Abstracts Head, Department of Immunogenetics and Cell Culture, Bu-Ali Research Institute (BARI), Department of Immunogenetics and Cell Culture, Mashhad, Iran, Rashin Ganjali, Researcher, Bu-Ali Research Institute (BARI), Department of Immunogenetics, Mashhad, Iran, Javad Ghaffari, Mazandaran University of Medical Sciences, Department of Pediatrics, Sari, Iran, Houshang Rafatpanah, Ghaem Medical Center, Department of Allergy and Immunology, Mashhad, Iran, Reza Farid-Hosseini, Head, Department of Allergy and Immunology, Ghaem Medical Center, Department of Allergy and Immunology, Mashhad, Iran Background: Idiopathic Chronic Urticaria (ICU), the most common form (70–80%) of chronic urticaria is supposed to have immune basis causes. It is speculated that the promoter polymorphism of TGF-β1 gene may be involved in ICU. This condition is thought to affect at least 0.1% of the population and often it can be severe and difficult to treat. Materials and Methods: A total of 40 patients with ICU and 41 normal subjects were studied. DNA was extracted from whole blood and TGF-β1 promoter − 509CNT polymorphism was determined by PCR-RFLP method. Results: Out of the 40 patients with ICU, 11 (27.5%) had CC, 26 (65%) had CTand 3 (7.5%) had TT genotypes. A higher proportion of case subjects with the C allele (CT type or CC type) was found compared with the T allele. Conclusion: These results do suggest an influence of genetic variability at the promoter of TGF-β1 gene (− 509CNT) on the occurrence of ICU. This polymorphism has been shown as a useful genetic change in our study. Further work is required to confirm this result. doi:10.1016/j.clim.2007.03.107

Su.67 Investigation of the Mechanisms of Pressure Ulcers Using a Cyclical Cutaneous Ischemic−reperfusion Injury Model Yuki Saito, Graduate Student, Kanazawa University, Dermatology, Kanazawa, Japan, Minoru Hasegawa, Assistant professor, Kanazawa University, Dermatology, Kanazawa, Japan, Manabu Fujimoto, Associate professor, Kanazawa university, Dermatology, Kanazawa, Japan, Kazuhiko Takehara, Professor, Kanazawa university, Dermatology, Kanazawa, Japan The formation of pressure ulcers is considered to be derived from multifactorial factors. Among them, the occurrence of cycles of ischemic–reperfusion is an especially significant contributing factor. This study assessed immunological mechanism of a reproducible murine model of ischemic–reperfusion injury by the external application of magnets. Dorsal skin was gently pulled and placed between two round ceramic magnetic plates. A single ischemic– reperfusion cycle (I–R cycle) consisted of a 12 hours of magnet placement followed by a 12 hours of release or rest. Three cycles were performed to induce pressure ulcer formation. A marked edema was observed after an I–R cycle (day 1) and skin ulcer developed until day 6 (3 days post treatment). Then, skin ulcers were healed until day 20 in all mice. In the lesional skin, the infiltration of neutrophils and macrophages, and tumor necrosis factor-alpha expression

Abstracts were most prominent at day 2. Then, inducible nitric oxide synthase (iNOS) expression was augmented at day 3. While nitric oxide is considered to be involved in ischemic– reperfusion injury, iNOS is a critical enzyme for the synthesis of nitric oxide. iNOS is produced by various cells including neutrophils and macrophages stimulated with proinflammatory cytokines such as tumor necrosis factor-alpha. Therefore, our findings suggest that iNOS production from infiltrated neutrophils and macrophages stimulated by tumor necrosis factor-alpha contributes to the development of skin ulcer in this model. We propose that this cyclical cutaneous ischemic–reperfusion injury model is useful for investigating the mechanism or developing novel therapies of pressure ulcers. doi:10.1016/j.clim.2007.03.109

Su.68 CD19 Expression in B cells is Important for Suppression of Contact Hypersensitivity Manabu Fujimoto, Associate Professor, Department of Dermatology, Kanzawa University, Kanazawa Ishikawa, Japan, Rei Watanabe, Student, Department of Dermatology, University of Tokyo, Bunkyo Tokyo, Japan, Thomas Tedder, Professor, Department of Immunology, Duke University Medical Center, Durham, NC, Kunihiko Tamaki, Professor, Department of Dermatology, University of Tokyo, Bunkyo Tokyo, Japan Contact hypersensitivity (CHS) is a cutaneous immune reaction mediated mainly by Ag-specific effector T cells, and regarded as a model for Th1/Tc1-mediated inflammation. However, recent reports have suggested pivotal roles of B cells in CHS. CD19 is a member of the Ig superfamily expressed on B cells and follicular dendritic cells, and serves as a positive B cell response regulator which defines signaling thresholds critical for B cell responses. In the current study, we assessed the role of the B cell-specific surface molecule CD19 on the development of CHS through examining CD19deficient mice. Although CD19-deficient mice are hyposensitive to a variety of transmembrane signals, CD19 loss resulted in increased and prolonged reaction of CHS, suggesting an inhibitory role of CD19 expression in the etiology of CHS. Sensitized draining lymph nodes and elicited ear lesions from CD19-deficient mice exhibited Th1/Tc1-shifted cytokine profile with increased IFN-fÁ expression and decreased IL10 expression. Adoptive transfer experiments revealed that CD19 expression in recipient mice was required for optimal suppression of CHS response, indicating its role in elicitation phase. Furthermore, spleen B-2 cells, especially marginal zone B cells, from wild type mice were able to normalize exaggerated CHS reactions in CD19-deficient mice. Thus, CD19 expression in B cells is critical for termination of CHS responses, possibly through the function of regulatory B cells. doi:10.1016/j.clim.2007.03.111

Su.69 Automated Enumeration of Skin Mast Cells by Laser Scanning Cytometry Esther Trueblood, Director of Pathology, Amgen, Inc./ Pathology, Seattle, WA, Stephen Z., Scientist, Amgen, Inc./

S165 Clinical Immunology, Thousand Oaks, CA, Andrea Ita, Senior Scientist, Amgen, Inc./Inflammation, Thousand Oaks, CA, John Ferbas, Director Medical Sciences, Amgen, Inc./ Clinical Immunology, Thousand Oaks, CA, James Chung, Medical Sciences Director, Amgen, Inc./Early development, Thousand Oaks, CA, Gordon Ng, Scientific Director, Amgen, Inc./Inflammation, Thousand Oaks, CA, Gloria Juan, Senior Scientist, Amgen, Inc./Clinical Immunology, Thousand Oaks, CA Mast cells (MCs) are bone marrow-derived immune competent cells dependent on stem cell factor for survival, chemotaxis, functional activation, and adhesion to connective tissue matrix. After activation, MCs can release a variety of mediators including histamine, tryptase, leukotrienes, prostaglandins, contributing to anaphylactoid, allergic and inflammatory processes. MCs are also associated with wound repair together with fibroblasts and other immune cells. The current effort presents a novel application of Laser Scanning Cytometry (LSC) to study the time course of mast cell recruitment in a skin woundhealing model. A full-thickness incisional wound model was developed in the mouse. A skin biopsy was taken on day 8 after wounding in animals treated with control or an antimurine c-Kit antibody (ACK2) that blocks SCF. A two-step protocol on the LSC was created to automate quantitation of wound-adjacent MCs. The first step executed a lowresolution optical scan that defined the position and area of each biopsy on the slide. The second step consisted of a high-resolution scan that captured sequential 20× image fields of the entire biopsy and stitched the images together into a mosaic. Computer analysis segregated stored image events into discrete populations, and ultimately MCs. MCs counts were obtained from a defined region around the wound site. ACK2 treatment reduced wound-activated Mast Cell expansion as measured by LSC (T-test; P = 0.0094) and consistent with the manual counts. This method of quantitation allows implementation of a more efficient, objective and precise method to score effects on MCs which are tissue resident and difficult to analyze. doi:10.1016/j.clim.2007.03.112

Su.70 Kawasaki Disease: Presentation of One Case Carlos Torres-Loza, Immunoallergist, West National Medical Center, UMAE-IMSS, Guadalajara, Mexico Kawasaki disease (KD) is the most common vasculitis of childhood and may well be the most common vasculitis at any age. The aim of this work is to present a case with an atypical clinical presentation or perhaps to consider other immunoinflamatory syndrome and to share this clinical experience with other immunologists. Ketzalli is a girl of 5 years old who started abruptly with abdominal pain, remittent fever often up 38.5 °C and scarlatiniform and multiform cutaneous rash and irritability by 2–3 days. After that acute period, we treated with metamizole as antithermic. After that period, we realized that our patient started to feel better but, with changes in her lips characterized by dry, swollen and vertically cracked lips and diffusely without strawberry tongue. Actually, by that time we realized both erythema of palms