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Evaluation of antioxidant enzyme levels as biological markers at different stages of pneumoconiosis in coal workers
Abstracts / Toxicology Letters 180S (2008) S32–S246
(p < 0.01) on peripheral blood cells in allergic children while expression of CD62L was not changed. Conclusions: In conclusion, permanent stimulation of immune system in allergic children might lead to unbalanced response of lymphocytes to stimuli. Acknowledgements: We would like to express our gratitude to Viera Vachalkova, Helena Turazova, Edita Mrvikova, Mikulas Krnac, Adriena Paulikova, Zuzana Kormancikova and Olga Liskova. The Ministry of Health of the Slovak Republic # 2005/40-SZU-18 and US NIH # 2 D43 TW00621-006 supported this work. doi:10.1016/j.toxlet.2008.06.147 I35 Evaluation of antioxidant enzyme levels as biological markers at different stages of pneumoconiosis in coal workers Ozge Cemiloglu Ulker 1,∗ , Berran Yucesoy 1 , Ozgur Demir 2 , Ishak Ozel Tekin 3 , Asuman Karakaya 1 1
Ankara University, Faculty of Pharmacy, Department of Toxicology, Ankara, Turkey, 2 Zonguldak Chest and Occupational Disease Hospital, Zonguldak, Turkey, 3 Karaelmas University, Faculty of Medicine, Department of Immunology, Zonguldak, Turkey Coal workers’ pneumoconiosis (CWP) which is an occupational pulmonary disease that occurs by chronic inhalation of coal dust is divided into two stages depending on the extent of the disease as simple pneumoconiosis (SP) and progressive massive fibrosis (PMF). Coal dust particles are known to stimulate the macrophages that produce excessive amounts of reactive oxygen species (ROS) which can be considered the mediators of chronic tissue damage and fibrosis. An imbalance between ROS and antioxidant enzymes could be related to the initiation and progression of CWP. Such an imbalance could be earliest detectable manifestation of progression from an asymptomatic preclinical stage to a disease severity. Also bronchoalveolar lavage (BAL) fluid would more closely reflect the in vivo imbalance of these mediators. In the present study we aimed to investigate serum and bronchoalveolar lavage antioxidant enzyme (SOD, GPx, catalase) levels in CWP patients and their relations with the disease severity. We found significantly elevated serum SOD levels, BAL SOD, GPx, catalase levels in SP and PMF patient groups. Also nonsignificant increases were observed in BAL antioxidant enzyme levels of PMF patient group when compared to SP group. Based on these results, serum SOD and BAL antioxidant enzymes levels were evaluated and discussed as biomarkers of CWP. Acknowledgements: This study was supported by Ankara University Research Fund (Project no. 20030803036). doi:10.1016/j.toxlet.2008.06.148 I36 Measurement of immunological parameters in blood of Gottingen Minipigs® and the T cell-dependent immune response to KLH Geertje van Mierlo ∗ , Marcel Schijf, Mary-Lène de Zeeuw-Brouwer, Jolanda van Bilsen, André Penninks TNO Quality of Life, Zeist, Netherlands To support human clinical trials with new pharmaceuticals, the harmonized guidelines for immunotoxicity evaluation (EMEA/CHMP/167235/2004-ICH) suggest testing new substances
S219
in at least two different species, a rodent and a non-rodent. There is growing interest to use minipigs as an alternative species to traditional non-rodent species for non-clinical studies of pharmaceuticals, especially non-human primates. Therefore, we developed experience to be able to study the desired and undesired effects of drugs and biopharmaceuticals on the immune system in minipigs. In an in vivo study with Göttingen minipigs® we studied different immunological endpoints including the T cell-dependent antibody response to Keyhole Limpet Hemocyanin (KLH). Results will be presented on the primary and secondary KLH response in minipigs that were either injected with vehicle control or KLH on days 0 and 14 [primary and secondary KLH-specific IgM and IgG levels and the Delayed Type Hypersensitivity (DTH) against KLH]. Moreover, at different time points, blood samples were taken and analyzed for different additional immunological endpoints, i.e. Natural Killer (NK) cell activity, lymphocyte subset analysis and proliferation of Peripheral Blood Mononuclear cells (PBMC) after in vitro stimulation with Concanavalin A or KLH. Currently, a sub-acute (38 days) immunotoxicity study in Göttingen minipigs® is running in which the animals are treated with either vehicle or immune modulators. In these animals, the immunological endpoints in response to KLH-immunization, as successfully implemented, will be followed in time. Preliminary results of this study will also be presented. doi:10.1016/j.toxlet.2008.06.149 I37 Cross-reactivity between trimellitic anhydride (TMA) and phthalic anhydride (PA) in respiratory allergy? Jos van Triel ∗ , Josje Arts, Hans Muijser, Gerard Roverts, Frieke Kuper TNO Quality of Life, Division Toxicology and Applied Pharmacology, Zeist, Netherlands An animal model for respiratory allergy was used to investigate the possible cross-reactivity between closely related allergens. Previously, we demonstrated that dermal sensitization with trimellitic anhydride (TMA), a well-known low molecular weight (LMW) respiratory allergen, induced elevated plasma IgE levels in Brown Norway (BN) rats. Subsequently, a respiratory challenge with TMA triggered functional and histopathological airway reactions characteristic of respiratory allergy. In the present study, BN rats (6 animals per group) were sensitized with TMA or the chemically related allergen phthalic anhydride (PA) on the shaved flank (day 0) and on the dorsum of both ears (day 7). Breathing pattern (frequency and tidal volume) was analysed before, during, directly after and 24 h after a respiratory challenge with either 12 mg/m3 or 35 mg/m3 TMA for 15 min on day 21. One day after challenge, bronchoalveolar lavage (BAL) was performed to determine total protein, lactate dehydrogenase, N-acetyl glucosaminidase, gamma-glutamyl-transferase, alkaline phosphatase and total and differential leukocyte numbers in the fluid. In addition, plasma IgE levels were analysed and the respiratory tract was collected for histopathological examination. Allergic airway reactions upon challenge with TMA were only observed in TMA-sensitized rats, not in PA- or sham-sensitized animals. Furthermore, none of the other investigated parameters demonstrated evidence for the presence of cross-reactivity between TMA and PA. If this observed absence of cross-reactivity is a general property, then – in the context of workplace safety – summation of the concentrations of even closely
(p < 0.01) on peripheral blood cells in allergic children while expression of CD62L was not changed. Conclusions: In conclusion, permanent stimulation of immune system in allergic children might lead to unbalanced response of lymphocytes to stimuli. Acknowledgements: We would like to express our gratitude to Viera Vachalkova, Helena Turazova, Edita Mrvikova, Mikulas Krnac, Adriena Paulikova, Zuzana Kormancikova and Olga Liskova. The Ministry of Health of the Slovak Republic # 2005/40-SZU-18 and US NIH # 2 D43 TW00621-006 supported this work. doi:10.1016/j.toxlet.2008.06.147 I35 Evaluation of antioxidant enzyme levels as biological markers at different stages of pneumoconiosis in coal workers Ozge Cemiloglu Ulker 1,∗ , Berran Yucesoy 1 , Ozgur Demir 2 , Ishak Ozel Tekin 3 , Asuman Karakaya 1 1
Ankara University, Faculty of Pharmacy, Department of Toxicology, Ankara, Turkey, 2 Zonguldak Chest and Occupational Disease Hospital, Zonguldak, Turkey, 3 Karaelmas University, Faculty of Medicine, Department of Immunology, Zonguldak, Turkey Coal workers’ pneumoconiosis (CWP) which is an occupational pulmonary disease that occurs by chronic inhalation of coal dust is divided into two stages depending on the extent of the disease as simple pneumoconiosis (SP) and progressive massive fibrosis (PMF). Coal dust particles are known to stimulate the macrophages that produce excessive amounts of reactive oxygen species (ROS) which can be considered the mediators of chronic tissue damage and fibrosis. An imbalance between ROS and antioxidant enzymes could be related to the initiation and progression of CWP. Such an imbalance could be earliest detectable manifestation of progression from an asymptomatic preclinical stage to a disease severity. Also bronchoalveolar lavage (BAL) fluid would more closely reflect the in vivo imbalance of these mediators. In the present study we aimed to investigate serum and bronchoalveolar lavage antioxidant enzyme (SOD, GPx, catalase) levels in CWP patients and their relations with the disease severity. We found significantly elevated serum SOD levels, BAL SOD, GPx, catalase levels in SP and PMF patient groups. Also nonsignificant increases were observed in BAL antioxidant enzyme levels of PMF patient group when compared to SP group. Based on these results, serum SOD and BAL antioxidant enzymes levels were evaluated and discussed as biomarkers of CWP. Acknowledgements: This study was supported by Ankara University Research Fund (Project no. 20030803036). doi:10.1016/j.toxlet.2008.06.148 I36 Measurement of immunological parameters in blood of Gottingen Minipigs® and the T cell-dependent immune response to KLH Geertje van Mierlo ∗ , Marcel Schijf, Mary-Lène de Zeeuw-Brouwer, Jolanda van Bilsen, André Penninks TNO Quality of Life, Zeist, Netherlands To support human clinical trials with new pharmaceuticals, the harmonized guidelines for immunotoxicity evaluation (EMEA/CHMP/167235/2004-ICH) suggest testing new substances
S219
in at least two different species, a rodent and a non-rodent. There is growing interest to use minipigs as an alternative species to traditional non-rodent species for non-clinical studies of pharmaceuticals, especially non-human primates. Therefore, we developed experience to be able to study the desired and undesired effects of drugs and biopharmaceuticals on the immune system in minipigs. In an in vivo study with Göttingen minipigs® we studied different immunological endpoints including the T cell-dependent antibody response to Keyhole Limpet Hemocyanin (KLH). Results will be presented on the primary and secondary KLH response in minipigs that were either injected with vehicle control or KLH on days 0 and 14 [primary and secondary KLH-specific IgM and IgG levels and the Delayed Type Hypersensitivity (DTH) against KLH]. Moreover, at different time points, blood samples were taken and analyzed for different additional immunological endpoints, i.e. Natural Killer (NK) cell activity, lymphocyte subset analysis and proliferation of Peripheral Blood Mononuclear cells (PBMC) after in vitro stimulation with Concanavalin A or KLH. Currently, a sub-acute (38 days) immunotoxicity study in Göttingen minipigs® is running in which the animals are treated with either vehicle or immune modulators. In these animals, the immunological endpoints in response to KLH-immunization, as successfully implemented, will be followed in time. Preliminary results of this study will also be presented. doi:10.1016/j.toxlet.2008.06.149 I37 Cross-reactivity between trimellitic anhydride (TMA) and phthalic anhydride (PA) in respiratory allergy? Jos van Triel ∗ , Josje Arts, Hans Muijser, Gerard Roverts, Frieke Kuper TNO Quality of Life, Division Toxicology and Applied Pharmacology, Zeist, Netherlands An animal model for respiratory allergy was used to investigate the possible cross-reactivity between closely related allergens. Previously, we demonstrated that dermal sensitization with trimellitic anhydride (TMA), a well-known low molecular weight (LMW) respiratory allergen, induced elevated plasma IgE levels in Brown Norway (BN) rats. Subsequently, a respiratory challenge with TMA triggered functional and histopathological airway reactions characteristic of respiratory allergy. In the present study, BN rats (6 animals per group) were sensitized with TMA or the chemically related allergen phthalic anhydride (PA) on the shaved flank (day 0) and on the dorsum of both ears (day 7). Breathing pattern (frequency and tidal volume) was analysed before, during, directly after and 24 h after a respiratory challenge with either 12 mg/m3 or 35 mg/m3 TMA for 15 min on day 21. One day after challenge, bronchoalveolar lavage (BAL) was performed to determine total protein, lactate dehydrogenase, N-acetyl glucosaminidase, gamma-glutamyl-transferase, alkaline phosphatase and total and differential leukocyte numbers in the fluid. In addition, plasma IgE levels were analysed and the respiratory tract was collected for histopathological examination. Allergic airway reactions upon challenge with TMA were only observed in TMA-sensitized rats, not in PA- or sham-sensitized animals. Furthermore, none of the other investigated parameters demonstrated evidence for the presence of cross-reactivity between TMA and PA. If this observed absence of cross-reactivity is a general property, then – in the context of workplace safety – summation of the concentrations of even closely