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Mast Cell Activation by Antigen Promotes Expression of IL-33
Abstracts S109
J ALLERGY CLIN IMMUNOL VOLUME 121, NUMBER 2
419
An Academic Medical Center-Based HIV Educational and Quality of Life Network Created by/for HIV-infected Youth Survivors who Contribute to their Community and the Medical Profession L. M. Noroski1,2, T. M. Aldape1,2, H. A. Anonymous2; 1Baylor College of Medicine, Houston, TX, 2Kids Council-Caregivers League, Houston, TX. RATIONALE: Perinatally HIV-infected adults have survived via effective antiretrovirals; however, their well-being is challenged by prevailing HIV stigma - this was not foreseen. Surviving HIV-infected youth have the potential to contribute to themselves/society through linking to their medical center in ways that would otherwise not be satisfied. Standardized approaches to HIV education/well-being linked to traditional medical care do not exist. Academic medical centers have the infrastructure to extend HIV care to population-specific HIV education/well-being programming and HIV youth can gain facilitation to sustain their own program. METHODS: Needs Assessment/Preceed: perinatally HIV-infected teens; medical/social/public health professionals; highly-charged HIV issues focus groups target ’’kids helping other kids’’; expanded to families/ medical/greater communities. Youth/family learn facts from health professionals then co-author HIV curriculum to teach communities and/or become hospital volunteers. HIV anonymity codified:authorship name - none:full; some:pen; complete:none. RESULTS: Founding HIV-infected youth created an HIV educational network to initially help themselves that multiplied (>53) with continued increase and remains sustainable (5 years); it grew into a unique patientinfluenced HIV sounding board for medical/public health in-trainees/ professionals with HIV youth as authors/teachers/volunteers (6:200 works:satisfactory encounters/year) and established community collaborations (3:1 local:international). CONCLUSIONS: Complex HIV issues drove surviving perinatally HIVinfected teens to create their own educational network linked to guidance/ support from their academic center. They landed career development stepping stones for themselves while contributing to a unique HIV educational forum that serves their medical/greater communities and strives to minimize HIV stigma. This patient-founded entity may serve as a contributing model to the larger development of much needed hospitalto-society HIV education/well-being standards. Funding: Texas Children’s Hospital
420
Environmental Stresses on Parents of Food Allergic Children can be Measured by Implicit Association Testing in the Clinic D. DunnGalvin, A. DunnGalvin, J. O. B. Hourihane; University College Cork, Cork, IRELAND. RATIONALE: Parental perception of childhood food allergy (FA) has a significant impact on both parents and children. Self-report measures are subject to bias. The implicit association test (IAT) measures unconscious associations, using relevant word pairs flashed onto a computer screen, timing the interval to key pressing. Longer times represent weaker associations between the presented concepts. IAT is not susceptible to self-reporting bias. We piloted measurement of the association between environmental stimuli (e.g. home, school, party) and anxiety in parents of FA children using IAT and a questionnaire. METHOD: 30 parents of FA children completed an IAT and a self-report questionnaire. The IAT measured the differential association of a target category (environmental stimuli) with an attribute dimension (anxious vs. relaxed). A study specific questionnaire (SSQ) was used to explicitly measure parental anxiety relating to attitudes to their child’s environment. An effect can be demonstrated on IAT with a score of >0.2.
RESULTS: Effect size for the IAT was larger 1.02 (M 5 0.52, SD 5 0.51, p < .05) than for the questionnaire 0.31 (M 5 0.40, SD 5 1.27, p < .05). This shows IAT was more sensitive than the questionnaire. CONCLUSIONS: Our novel findings demonstrate that specific environmental stimuli or situations trigger high anxiety in parents of FA children. The ease of IAT testing will clarify and provide more depth to previous quantitative research findings in this area by assessing automatic (unconscious) cognitive processes with theoretical and clinical implications.
421
Increased Mast Cell Adhesion Molecule Immunoglobulin Super Family 4 (IGSF4) in Distal Lung Fibroblasts: Regulation by Transforming Growth Factor-ß2 M. G. Ghoshhajra, H. Z. Hu, J. B. Trudeau, S. E. Wenzel; University of Pittsburgh Medical Center, Pittsburgh, PA. RATIONALE: Mast cell numbers are higher in the distal lung. In severe asthma, high mast cell numbers are maintained in distal lung despite low numbers proximally. We hypothesized that differential expression of mast cell growth factors/adhesion molecules such as stem cell factor (SCF) and IGSF4 by proximal and distal lung fibroblasts could contribute to these differences. METHODS: 3rd to 4th passage endobronchial and transbronchial biopsy fibroblasts from mild (n 5 3) and severe (n 5 6) asthmatics were exposed to TGF-ß2 in both time course (n 5 7) and dose-response (n 5 3) experiments. Quantitative real-time PCR and Western blot were performed to measure IGSF4, while SCF was evaluated by real-time PCR only. RESULTS: Both proximal and distal lung fibroblasts expressed very low levels of SCF, which did not increase with TGF-ß2. In contrast, baseline IGSF4 expression was 2- to 270-fold higher in distal lung fibroblasts compared to proximal. TGF-ß2 increased IGSF4 in distal lung fibroblasts in both a time- and dose-dependent manner while having no effect on proximal lung fibroblasts. The maximum increase occurred between 4-24 hrs, and between 0.5 and 5 ng/ml of TGF-ß2. Western blot analysis confirmed higher basal and stimulated IGSF4 in distal lung fibroblasts than proximal. CONCLUSIONS: Increased expression of IGSF4 in distal asthmatic airways, especially after stimulation with TGF-ß2, could promote increased mast cell growth and survival. This may result in persistent distal lung mast cell inflammation in severe asthma. Funding: National Institutes of Health
422
Mast Cell Activation by Antigen Promotes Expression of IL-33 P. J. Bryce, C. V. Neilsen; Northwestern University, Chicago,
IL. RATIONALE: IL-33 is a recently identified member of the IL-1 family of cytokines. It has been shown to potently upregulate the expression of Th2 cytokines through its receptor ST2 and to enhance the survival and functions of mast cells. However, its specific cellular sources are not well characterized. Mast cell activation is also known to enhance Th2 cytokine responses and so we investigated if mast cells are a source of IL33. METHODS: Murine bone marrow derived mast cells (BMMC) were generated in vitro by culture with IL-3. Activation was achieved by administration of DNP-specific IgE for 24 hours, followed by addition of DNP-HSA. The expression of mRNA for IL-33 was determined at time points following antigen-driven activation. In addition, using intracellular flow cytometry, we studied the expression of IL-33 protein. RESULTS: Unactivated BMMC’s showed very low expression of IL-33 message. However, antigen-driven activation promoted a rapid and significant increase in IL-33 expression that peaked 4h after stimulation and resolved by 24h. Intracellular flow cytometry showed that by 6h after stimulation, more than 80% of BMMC’s were positive for IL-33 protein. CONCLUSIONS: Our findings establish that antigen-mediated activation via FceRI promotes the expression of IL-33 in mast cells. We are investigating the possibility that IL-33 might serve as an autocrine regulator of mast cell functions after activation and as a paracrine connection between mast cell activation and Th2 cytokine responses.
SUNDAY
categories was good (k 0.84). Internal consistency reliability was high, with Cronbach’s scale reliability coefficient 0.90 for Quolla-A and 0.93 for Quolla-B. CONCLUSIONS: Spanish Quolla version has been demonstrated to be feasible and reliable. Validation phase in a higher sample size is necessary, and it is been performed now.
J ALLERGY CLIN IMMUNOL VOLUME 121, NUMBER 2
419
An Academic Medical Center-Based HIV Educational and Quality of Life Network Created by/for HIV-infected Youth Survivors who Contribute to their Community and the Medical Profession L. M. Noroski1,2, T. M. Aldape1,2, H. A. Anonymous2; 1Baylor College of Medicine, Houston, TX, 2Kids Council-Caregivers League, Houston, TX. RATIONALE: Perinatally HIV-infected adults have survived via effective antiretrovirals; however, their well-being is challenged by prevailing HIV stigma - this was not foreseen. Surviving HIV-infected youth have the potential to contribute to themselves/society through linking to their medical center in ways that would otherwise not be satisfied. Standardized approaches to HIV education/well-being linked to traditional medical care do not exist. Academic medical centers have the infrastructure to extend HIV care to population-specific HIV education/well-being programming and HIV youth can gain facilitation to sustain their own program. METHODS: Needs Assessment/Preceed: perinatally HIV-infected teens; medical/social/public health professionals; highly-charged HIV issues focus groups target ’’kids helping other kids’’; expanded to families/ medical/greater communities. Youth/family learn facts from health professionals then co-author HIV curriculum to teach communities and/or become hospital volunteers. HIV anonymity codified:authorship name - none:full; some:pen; complete:none. RESULTS: Founding HIV-infected youth created an HIV educational network to initially help themselves that multiplied (>53) with continued increase and remains sustainable (5 years); it grew into a unique patientinfluenced HIV sounding board for medical/public health in-trainees/ professionals with HIV youth as authors/teachers/volunteers (6:200 works:satisfactory encounters/year) and established community collaborations (3:1 local:international). CONCLUSIONS: Complex HIV issues drove surviving perinatally HIVinfected teens to create their own educational network linked to guidance/ support from their academic center. They landed career development stepping stones for themselves while contributing to a unique HIV educational forum that serves their medical/greater communities and strives to minimize HIV stigma. This patient-founded entity may serve as a contributing model to the larger development of much needed hospitalto-society HIV education/well-being standards. Funding: Texas Children’s Hospital
420
Environmental Stresses on Parents of Food Allergic Children can be Measured by Implicit Association Testing in the Clinic D. DunnGalvin, A. DunnGalvin, J. O. B. Hourihane; University College Cork, Cork, IRELAND. RATIONALE: Parental perception of childhood food allergy (FA) has a significant impact on both parents and children. Self-report measures are subject to bias. The implicit association test (IAT) measures unconscious associations, using relevant word pairs flashed onto a computer screen, timing the interval to key pressing. Longer times represent weaker associations between the presented concepts. IAT is not susceptible to self-reporting bias. We piloted measurement of the association between environmental stimuli (e.g. home, school, party) and anxiety in parents of FA children using IAT and a questionnaire. METHOD: 30 parents of FA children completed an IAT and a self-report questionnaire. The IAT measured the differential association of a target category (environmental stimuli) with an attribute dimension (anxious vs. relaxed). A study specific questionnaire (SSQ) was used to explicitly measure parental anxiety relating to attitudes to their child’s environment. An effect can be demonstrated on IAT with a score of >0.2.
RESULTS: Effect size for the IAT was larger 1.02 (M 5 0.52, SD 5 0.51, p < .05) than for the questionnaire 0.31 (M 5 0.40, SD 5 1.27, p < .05). This shows IAT was more sensitive than the questionnaire. CONCLUSIONS: Our novel findings demonstrate that specific environmental stimuli or situations trigger high anxiety in parents of FA children. The ease of IAT testing will clarify and provide more depth to previous quantitative research findings in this area by assessing automatic (unconscious) cognitive processes with theoretical and clinical implications.
421
Increased Mast Cell Adhesion Molecule Immunoglobulin Super Family 4 (IGSF4) in Distal Lung Fibroblasts: Regulation by Transforming Growth Factor-ß2 M. G. Ghoshhajra, H. Z. Hu, J. B. Trudeau, S. E. Wenzel; University of Pittsburgh Medical Center, Pittsburgh, PA. RATIONALE: Mast cell numbers are higher in the distal lung. In severe asthma, high mast cell numbers are maintained in distal lung despite low numbers proximally. We hypothesized that differential expression of mast cell growth factors/adhesion molecules such as stem cell factor (SCF) and IGSF4 by proximal and distal lung fibroblasts could contribute to these differences. METHODS: 3rd to 4th passage endobronchial and transbronchial biopsy fibroblasts from mild (n 5 3) and severe (n 5 6) asthmatics were exposed to TGF-ß2 in both time course (n 5 7) and dose-response (n 5 3) experiments. Quantitative real-time PCR and Western blot were performed to measure IGSF4, while SCF was evaluated by real-time PCR only. RESULTS: Both proximal and distal lung fibroblasts expressed very low levels of SCF, which did not increase with TGF-ß2. In contrast, baseline IGSF4 expression was 2- to 270-fold higher in distal lung fibroblasts compared to proximal. TGF-ß2 increased IGSF4 in distal lung fibroblasts in both a time- and dose-dependent manner while having no effect on proximal lung fibroblasts. The maximum increase occurred between 4-24 hrs, and between 0.5 and 5 ng/ml of TGF-ß2. Western blot analysis confirmed higher basal and stimulated IGSF4 in distal lung fibroblasts than proximal. CONCLUSIONS: Increased expression of IGSF4 in distal asthmatic airways, especially after stimulation with TGF-ß2, could promote increased mast cell growth and survival. This may result in persistent distal lung mast cell inflammation in severe asthma. Funding: National Institutes of Health
422
Mast Cell Activation by Antigen Promotes Expression of IL-33 P. J. Bryce, C. V. Neilsen; Northwestern University, Chicago,
IL. RATIONALE: IL-33 is a recently identified member of the IL-1 family of cytokines. It has been shown to potently upregulate the expression of Th2 cytokines through its receptor ST2 and to enhance the survival and functions of mast cells. However, its specific cellular sources are not well characterized. Mast cell activation is also known to enhance Th2 cytokine responses and so we investigated if mast cells are a source of IL33. METHODS: Murine bone marrow derived mast cells (BMMC) were generated in vitro by culture with IL-3. Activation was achieved by administration of DNP-specific IgE for 24 hours, followed by addition of DNP-HSA. The expression of mRNA for IL-33 was determined at time points following antigen-driven activation. In addition, using intracellular flow cytometry, we studied the expression of IL-33 protein. RESULTS: Unactivated BMMC’s showed very low expression of IL-33 message. However, antigen-driven activation promoted a rapid and significant increase in IL-33 expression that peaked 4h after stimulation and resolved by 24h. Intracellular flow cytometry showed that by 6h after stimulation, more than 80% of BMMC’s were positive for IL-33 protein. CONCLUSIONS: Our findings establish that antigen-mediated activation via FceRI promotes the expression of IL-33 in mast cells. We are investigating the possibility that IL-33 might serve as an autocrine regulator of mast cell functions after activation and as a paracrine connection between mast cell activation and Th2 cytokine responses.
SUNDAY
categories was good (k 0.84). Internal consistency reliability was high, with Cronbach’s scale reliability coefficient 0.90 for Quolla-A and 0.93 for Quolla-B. CONCLUSIONS: Spanish Quolla version has been demonstrated to be feasible and reliable. Validation phase in a higher sample size is necessary, and it is been performed now.