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Identification of snow crab proteins that elicit IgE reactivity in snow crab processing workers
J ALLERGY CLIN IMMUNOL VOLUME 111, NUMBER 2
05
Quantitative Structural Activity Relationship and Animal Modeling Accurately Predict the Potential for Human Dermal Sensitization and the Induction of Airway Hyperreactivity Following 3-amino-5-mercapto-l,2,4-triazole (AMT) Exposure
K. J. Klink 1, E. Hnizdo 2, B. J. MeadO; IAIG/OD Group, National Institute for Occupational Safety and Health, Morgantown, WV, 2Division of Respiratory Disease Studies, National Institute for Occupational Safety and Health, Morgantown, WV. In a Health Hazard Evaluation conducted by NIOSH, exposure to 3amino-5-mercapto-l,2,4-triazole (AMT), a feedstock chemical, and the resultant herbicide, AMT based product 1 (AMT-bpl) were associated with an occurrence of a cluster of work-related asthma. Computer and animal modeling were used to evaluate the potential for AMT and/or AMTbp 1 to induce dermal sensitization and airway hyperreactivity. Utilizing TOPKAT quantitative structural activity relationship modeling and the Local Lymph Node Assay, AMT, but not AMT-bp 1, was positive for dermal sensitization potential. No signs of systemic toxicity were observed in BALB/c mice following 28 days dermal exposure with concentrations up to 25% AMT. AMT was negative for contact hypersensitivity in the mouse ear swelling test up to concentrations of 25% AMT. Analysis of draining lymph node cells demonstrated an increase in IgE+B220+ cells by phenotypic analysis and an increase in IL-4 mRNA by RT-PCR, suggesting a Th2 dependent mechanism of sensitization. Following dermal exposure for 21 continuous days, animals exposed to 15% AMT demonstrated elevated levels of total serum IgE and an increase in airway hyperreactivity upon methacholine challenge. These studies demonstrate that exposure to AMT but not AMT based product 1 has the potential to induce sensitization and airway hyperreactivity following dermal exposure and may be responsible for the increase incidence of occupational asthma in the herbicide factory. Funding: Grant Monies. These studies were supported in part by NIEHS lAG No. Y1-ES-O049-03.
106 Identification of Snow Crab Proteins That Elicit IgE Reactivity in Snow Crab Processing Workers B. V. Gill I, A. Cartier 2, L. Horth-Susin 3, M. Jong 4, M. Swanson 5, B. Neis 6, S. B. LehrerT; tSection of Clinical Immunology, Allergy, and Rheumatology, Tulane University Health Sciences Center, New Orleans, LA, 2H. du Sacre-Coeur de Montreal, Montreal, PQ, CANADA, 3Valley Regional Hospital, Kentville, NS, CANADA, 4Health Labrador Corporation, Goose Bay, NS, CANADA, 5Mayo Clinic, Rochester, MN, 6Memorial University, St-John's, NF, CANADA, 7Tulane University Health Sciences Center, New Orleans, LA. RATIONALE: Snow crab proteins are known to produce lgE-mediated reactions in processing workers. This study sought to identify the relevant allergenic proteins. METHODS: The 24 most highly IgE-reactive sera were identified from 214 snow crab workers by radioallergosorbent test (RAST). Snow crab proteins in meat and cooking water extracts were separated by SDS-PAGE electrophoresis then transferred to nitrocellulose membranes. The membranes were incubated sequentially with the IgE-positive sera then 15,000 cpm/strip of 125I goat-anti-human IgE. The membranes were then exposed to autoradiography film and the resultant bands detected on the film were used to identify the IgE-reactive proteins. RESULTS: Of the 24 sera immunoblotted, 22 demonstrated lgE binding to multiple proteins weighing 13.6 kd to 50 kd. A majority of the sera reacted to 3 proteins in both extracts: a 34.3 kd protein, 79% sera reacted to the meat extract and 71% to the water extract, an 18.5 kd protein, 66% (meat) and 63% (water), and a 14.4 kd protein, 75% (meat and water). A fourth 25 kd protein elicited reactivity more frequently to the meat extract, 79%, than to the water extract, 33%. CONCLUSIONS: Three major lgE-reactive proteins were identified in snow crab meat and cooking water, with reactivity to a fourth protein
Abstracts
$97
being more prevalent in the snow crab meat. Identification of these proteins will aid in diagnosis of snow crab allergy and allow processing plants to identify the offending proteins in the work environment and develop methods to minimize them, ultimately reducing sensitization in the workplace. Funding: Partly supported by CIHR as part of Safety Net
07 Occupational Allergy due to Trichoplusia Ni Moth M. de las Heras, J. Cuesta, J. de Miguel, C. Escudero, M. Fern~indezNieto, C. Pastor, E Vivanco, S. Quirce, J. Sastre; Fundacion Jimenez Diaz, Madrid, SPAIN. RATIONALE: Trichoplusia ni (TN) is a worldwide distributed insect belonging to Lepidopterae order. The TN caterpillar (cabbage looper) is a serious pest of crucifers and other cultivated plants which is also used in medical research. CASE REPORT: We report a 27-year-old woman, who has been working as a biologist with TN moths for three years. Two years after starting her job she developed work-related symptoms of rhinoconjunctivitis and episodes of cough on days when she handled the moths and stayed in the facility where these insects are bred. METHODS AND RESULTS: Baseline spirometry was normal and the methacholine PC20 was 1.37 mg/ml. Extracts with TN larvae (6.7 mg/ml) and adults (5.4 mg/ml) were prepared. Skin prick tests (SPT) were positive to pollens, Anisakis simplex, and also to the larva and adult extracts eliciting a wheal of 7 mm and 10 mm, respectively. An specific nasal challenge test (SNC) performed with TN larva extract (0.006 mg/ml) measured by acoustic rhinometry caused an immediate 53% fall in minimum cross-sectional area. Specific IgE antibodies by ELISA technique were strongly positive (larva 0.95 OD, adult moth 0.66 OD, control 0.18 OD). IgE immunoblotting revealed some reactive bands in the adult extract with a 66 kD allergen also present in the larva extract. CONCLUSIONS: To the best of our knowledge this is the first case of IgE-mediated occupational rhinoconjunctivitis caused by TN moth and demonstrated by SPT, specific IgE response and SNC test. Funding: Self-funded
08
Carmine Allergy, a Hidden Cause for Anaphylaxis?
E L. Lin; Allergy Clinic, Naval Medical Center, San Diego, CA. RATIONALE: Carmine, a naturally derived red dye, extracted from cochineal beetle has been used in the coloring of beverages, foods, medicines and cosmetics for years. It is generally regarded as safe, but rarely allergic reactions following ingestion have been reported. Three additional patients are reported here with anaphylaxis after consumption of various foods and beverages containing carmine. METHODS: The patients were prick skin tested to suspected foods and various ingredients. Carmine was used for skin testing at a concentration of 0.035%. A review of the literature on carmine allergy was also conducted. RESULTS: The patients were all women age 31-46. Incriminated foods included yogurt, ruby red grapefruit drink, imitation crab and candy. Two volunteered a history of itchy rash after applying red eye shadow. Skin tests in all 3 patients were strongly positive to carmine and negative to other foods and ingredients. Eleven other cases of allergic reaction to ingested carmine have been reported to date. Again, all were women. Most of these had a history of atopy and/or a history of a prior reaction to cosmetic use. CONCLUSION: Carmine allergy by ingestion may be more common than previously thought, especially in atopic women sensitized by exposure to carmine in cosmetics. Carmine should be considered and tested for in any allergic reaction after a meal potentially containing red dye. The exclusive involvement of women, and the specific history of reactions to use of cosmetics in some, suggests that exposure to cosmetics may be a common route of carmine sensitization.
05
Quantitative Structural Activity Relationship and Animal Modeling Accurately Predict the Potential for Human Dermal Sensitization and the Induction of Airway Hyperreactivity Following 3-amino-5-mercapto-l,2,4-triazole (AMT) Exposure
K. J. Klink 1, E. Hnizdo 2, B. J. MeadO; IAIG/OD Group, National Institute for Occupational Safety and Health, Morgantown, WV, 2Division of Respiratory Disease Studies, National Institute for Occupational Safety and Health, Morgantown, WV. In a Health Hazard Evaluation conducted by NIOSH, exposure to 3amino-5-mercapto-l,2,4-triazole (AMT), a feedstock chemical, and the resultant herbicide, AMT based product 1 (AMT-bpl) were associated with an occurrence of a cluster of work-related asthma. Computer and animal modeling were used to evaluate the potential for AMT and/or AMTbp 1 to induce dermal sensitization and airway hyperreactivity. Utilizing TOPKAT quantitative structural activity relationship modeling and the Local Lymph Node Assay, AMT, but not AMT-bp 1, was positive for dermal sensitization potential. No signs of systemic toxicity were observed in BALB/c mice following 28 days dermal exposure with concentrations up to 25% AMT. AMT was negative for contact hypersensitivity in the mouse ear swelling test up to concentrations of 25% AMT. Analysis of draining lymph node cells demonstrated an increase in IgE+B220+ cells by phenotypic analysis and an increase in IL-4 mRNA by RT-PCR, suggesting a Th2 dependent mechanism of sensitization. Following dermal exposure for 21 continuous days, animals exposed to 15% AMT demonstrated elevated levels of total serum IgE and an increase in airway hyperreactivity upon methacholine challenge. These studies demonstrate that exposure to AMT but not AMT based product 1 has the potential to induce sensitization and airway hyperreactivity following dermal exposure and may be responsible for the increase incidence of occupational asthma in the herbicide factory. Funding: Grant Monies. These studies were supported in part by NIEHS lAG No. Y1-ES-O049-03.
106 Identification of Snow Crab Proteins That Elicit IgE Reactivity in Snow Crab Processing Workers B. V. Gill I, A. Cartier 2, L. Horth-Susin 3, M. Jong 4, M. Swanson 5, B. Neis 6, S. B. LehrerT; tSection of Clinical Immunology, Allergy, and Rheumatology, Tulane University Health Sciences Center, New Orleans, LA, 2H. du Sacre-Coeur de Montreal, Montreal, PQ, CANADA, 3Valley Regional Hospital, Kentville, NS, CANADA, 4Health Labrador Corporation, Goose Bay, NS, CANADA, 5Mayo Clinic, Rochester, MN, 6Memorial University, St-John's, NF, CANADA, 7Tulane University Health Sciences Center, New Orleans, LA. RATIONALE: Snow crab proteins are known to produce lgE-mediated reactions in processing workers. This study sought to identify the relevant allergenic proteins. METHODS: The 24 most highly IgE-reactive sera were identified from 214 snow crab workers by radioallergosorbent test (RAST). Snow crab proteins in meat and cooking water extracts were separated by SDS-PAGE electrophoresis then transferred to nitrocellulose membranes. The membranes were incubated sequentially with the IgE-positive sera then 15,000 cpm/strip of 125I goat-anti-human IgE. The membranes were then exposed to autoradiography film and the resultant bands detected on the film were used to identify the IgE-reactive proteins. RESULTS: Of the 24 sera immunoblotted, 22 demonstrated lgE binding to multiple proteins weighing 13.6 kd to 50 kd. A majority of the sera reacted to 3 proteins in both extracts: a 34.3 kd protein, 79% sera reacted to the meat extract and 71% to the water extract, an 18.5 kd protein, 66% (meat) and 63% (water), and a 14.4 kd protein, 75% (meat and water). A fourth 25 kd protein elicited reactivity more frequently to the meat extract, 79%, than to the water extract, 33%. CONCLUSIONS: Three major lgE-reactive proteins were identified in snow crab meat and cooking water, with reactivity to a fourth protein
Abstracts
$97
being more prevalent in the snow crab meat. Identification of these proteins will aid in diagnosis of snow crab allergy and allow processing plants to identify the offending proteins in the work environment and develop methods to minimize them, ultimately reducing sensitization in the workplace. Funding: Partly supported by CIHR as part of Safety Net
07 Occupational Allergy due to Trichoplusia Ni Moth M. de las Heras, J. Cuesta, J. de Miguel, C. Escudero, M. Fern~indezNieto, C. Pastor, E Vivanco, S. Quirce, J. Sastre; Fundacion Jimenez Diaz, Madrid, SPAIN. RATIONALE: Trichoplusia ni (TN) is a worldwide distributed insect belonging to Lepidopterae order. The TN caterpillar (cabbage looper) is a serious pest of crucifers and other cultivated plants which is also used in medical research. CASE REPORT: We report a 27-year-old woman, who has been working as a biologist with TN moths for three years. Two years after starting her job she developed work-related symptoms of rhinoconjunctivitis and episodes of cough on days when she handled the moths and stayed in the facility where these insects are bred. METHODS AND RESULTS: Baseline spirometry was normal and the methacholine PC20 was 1.37 mg/ml. Extracts with TN larvae (6.7 mg/ml) and adults (5.4 mg/ml) were prepared. Skin prick tests (SPT) were positive to pollens, Anisakis simplex, and also to the larva and adult extracts eliciting a wheal of 7 mm and 10 mm, respectively. An specific nasal challenge test (SNC) performed with TN larva extract (0.006 mg/ml) measured by acoustic rhinometry caused an immediate 53% fall in minimum cross-sectional area. Specific IgE antibodies by ELISA technique were strongly positive (larva 0.95 OD, adult moth 0.66 OD, control 0.18 OD). IgE immunoblotting revealed some reactive bands in the adult extract with a 66 kD allergen also present in the larva extract. CONCLUSIONS: To the best of our knowledge this is the first case of IgE-mediated occupational rhinoconjunctivitis caused by TN moth and demonstrated by SPT, specific IgE response and SNC test. Funding: Self-funded
08
Carmine Allergy, a Hidden Cause for Anaphylaxis?
E L. Lin; Allergy Clinic, Naval Medical Center, San Diego, CA. RATIONALE: Carmine, a naturally derived red dye, extracted from cochineal beetle has been used in the coloring of beverages, foods, medicines and cosmetics for years. It is generally regarded as safe, but rarely allergic reactions following ingestion have been reported. Three additional patients are reported here with anaphylaxis after consumption of various foods and beverages containing carmine. METHODS: The patients were prick skin tested to suspected foods and various ingredients. Carmine was used for skin testing at a concentration of 0.035%. A review of the literature on carmine allergy was also conducted. RESULTS: The patients were all women age 31-46. Incriminated foods included yogurt, ruby red grapefruit drink, imitation crab and candy. Two volunteered a history of itchy rash after applying red eye shadow. Skin tests in all 3 patients were strongly positive to carmine and negative to other foods and ingredients. Eleven other cases of allergic reaction to ingested carmine have been reported to date. Again, all were women. Most of these had a history of atopy and/or a history of a prior reaction to cosmetic use. CONCLUSION: Carmine allergy by ingestion may be more common than previously thought, especially in atopic women sensitized by exposure to carmine in cosmetics. Carmine should be considered and tested for in any allergic reaction after a meal potentially containing red dye. The exclusive involvement of women, and the specific history of reactions to use of cosmetics in some, suggests that exposure to cosmetics may be a common route of carmine sensitization.