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Innate immunity response to selected cyanobacterial peptides
Abstracts / Toxicology Letters 238S (2015) S56–S383
age-matched controls, employed in office tasks. Plasma levels of IL-1, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, IL-12p70, TNF-␣, TNF- and INF-␥ from exposed and control workers were determined by flowcytometry with a Coulter Epics XL-MCL using the FlowCy-tomix Human Th1/Th2 11plex Kit (Bender MedSystems,Vienna, Austria). Urinary levels of 3-phenoxybenzoic acid (3-PBA), a metabolite of ␣cypermethrin, were also determined to assess actual exposure level of each subject. Results: Urinary 3-PBA levels in exposed workers were significantly (p < 0.01) higher than controls, confirming actual pyrethroid absorption in the exposed group. The mean plasma IL-1b, IL-4, IL-5, IL-6, IL-10, TNF-␣ and TNF- levels were not significantly different between the two groups of workers in the study. Mean plasma levels of cytokines IL-12p70, INF-␥, IL-2 and IL-8 were significantly higher in exposed subjects. Nontheless, no correlation was found between urinary 3-PBA levels and plasma levels of the detected cytokines. Conclusions: The proinflammatory cytokines IL-2, IL-8, and IFN-␥ play a significant role in the defense against infection and cancer. These data on an exposed human population suggest that synthetic pyrethroids can induce immunotoxic effects, supporting the hypothesis that exposure to pyrethroids may increase the risk of infection and cancer. http://dx.doi.org/10.1016/j.toxlet.2015.08.658
P09-015 Innate immunity response to selected cyanobacterial peptides Z. Moosova 1,∗ , O. Adamovsky 1 , M. Pekarova 2 , L. Svihalkova Sindlerova 2 , L. Kubala 2 , L. Blaha 2 1 Masaryk University, Research Centre for Toxic Compounds in the Environment (RECETOX), Brno, Czech Republic 2 Academy of Sciences, Institute of Biophysics, Brno, Czech Republic
Cyanobacteria are ubiquitous photosynthesizing gram-negative bacteria frequently associated with toxic water bloom. Cyanobacterial production of a wide range of hepatotoxins, neurotoxins and dermatotoxins affects water quality, which is a growing environmental and health concern worldwide. Hepatotoxic microcystins (MCs) are very stable in the environment, and due to high risk to human health the WHO has established a provisional guideline of 1 g/L for MC-LR in drinking water. Up to now, only limited set of known cyanobacterial toxins were evaluated for immunomodulatory potencies. In our study, we have focused on characterization of the immunomodulation of MCs and aeruginosin in two experimental designs: (i) direct effects of cyanobacterial peptides on murine macrophage (RAW 264.7 cell line) viability and inflammatory response, (ii) and modulation of macrophage responses by cyanobacterial toxins in co-exposures with model stimuli (lipopolysaccharides, LPS). Cells were exposed to different concentrations of three MC congeners (1–1000 nM) and aeruginosin-865 (0.1–50 M) for 24 h. The production of nitric oxide (NO) in cell supernatants was quantified by Griess reagent method, cell viability was monitored using neutral red uptake assay and BCA protein assay kit, and the levels of pro-inflammatory cytokines (such as interleukin IL-6 and tumor necrosis factor TNF␣) were detected by ELISA method. Our results show that observed effects are congenerspecific. MC-LR was able to significantly affect the activation of macrophages, however, other most common MCs (-RR, -YR) and aeruginosin-865 did not alter immune responds. MC-LR also promoted production of NO and cytokines in co-exposure with LPS used as an established positive control. These results suggest that MC-LR may have significant immunomodulatory activities in very low environmentally relevant concentrations. The research was
S223
supported by the grant from the Czech National Science Foundation ˇ GP13-27695P). (GACR http://dx.doi.org/10.1016/j.toxlet.2015.08.659
P09-016 Genetic polymorphisms in the cytokine genes and risk of type 2 diabetes and its complications in a group of Turkish population A. Karakaya 1,∗ , I. Ates 1 , A. Güvenc¸ 2 , D. Altuner 3 1 Ankara University, Faculty of Pharmacy, Department of Toxicology, Ankara, Turkey 2 Kırsehir City Hospital, Kırsehir, Turkey 3 Erzincan University, Faculty of Medicine, Pharmacology, Erzincan, Turkey
Aim: Type 2 diabetes (Diabetes Mellitus, DM) is a hyperglycaemic metabolic disease related with carbohydrate, protein and lipid metabolism dysfunction due to the decrease of insulin secretion. Several genetic and environmental factors are playing major role in the development of the diabetic complications. Relationship between the inflammation generation and diabetic complications has been showed in recent studies. Following the formation of the oxidative stress after the disorder of the lipid metabolism, the levels of the reactive oxygen species (ROS) increase and insulin resistance develops consequently. Cytokines are important in regulation of the homeostatic mechanisms such as inflammation and tissue repair. Thus, variations in their levels and structures are the reasons for the occurrence of various diseases. The single nucleotide polymorphisms (SNP) forming on the cytokine genes increase the risk of disease development. Recent studies showed the relationship between some inflammatory cytokine gene polymorphisms and the development of complication in patients with diabetes. Based on this, our aim was searching and the evaluating the possible relations between the TNF-␣ (-308), IL-1 (+3953), IL-6 (-174) gene polymorphisms and the development of the complications in a Type 2 diabetic Turkish patient population. Methods: The study was performed by using PCR-RFLP method. Results and conclusions: In regards to our data, both TNF-␣ and IL-1 gene polymorphisms are significantly related with the development of both disease (3,27 fold and 2,15 fold respectively) and complications (4,15 fold and 2,11 fold respectively). On the other hand, there is a little but not significant effect of IL-6 gene polymorphism on the disease occurrence and complication development. http://dx.doi.org/10.1016/j.toxlet.2015.08.660
P09-017 Assessment of sensitization and photosensitization of avobenzone based on DPRA C. Benevenuto ∗ , L. Gaspar FCFRP-USP, Pharmaceutical Sciences, RIBEIRÃO PRETO, Brazil Purpose: Contact allergies are considered complex diseases it reaches 15–20% of the population. Among the cosmetic allergens, UV-filters have been considered emerging allergens and avobenzone (AVO) is one of the most used UVA-filter. The commonly methods to assess skin sensitization still use animals and among the few non-animal methods is the DPRA (direct peptide reactivity assay), which considers that if a substance can react with skin
age-matched controls, employed in office tasks. Plasma levels of IL-1, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, IL-12p70, TNF-␣, TNF- and INF-␥ from exposed and control workers were determined by flowcytometry with a Coulter Epics XL-MCL using the FlowCy-tomix Human Th1/Th2 11plex Kit (Bender MedSystems,Vienna, Austria). Urinary levels of 3-phenoxybenzoic acid (3-PBA), a metabolite of ␣cypermethrin, were also determined to assess actual exposure level of each subject. Results: Urinary 3-PBA levels in exposed workers were significantly (p < 0.01) higher than controls, confirming actual pyrethroid absorption in the exposed group. The mean plasma IL-1b, IL-4, IL-5, IL-6, IL-10, TNF-␣ and TNF- levels were not significantly different between the two groups of workers in the study. Mean plasma levels of cytokines IL-12p70, INF-␥, IL-2 and IL-8 were significantly higher in exposed subjects. Nontheless, no correlation was found between urinary 3-PBA levels and plasma levels of the detected cytokines. Conclusions: The proinflammatory cytokines IL-2, IL-8, and IFN-␥ play a significant role in the defense against infection and cancer. These data on an exposed human population suggest that synthetic pyrethroids can induce immunotoxic effects, supporting the hypothesis that exposure to pyrethroids may increase the risk of infection and cancer. http://dx.doi.org/10.1016/j.toxlet.2015.08.658
P09-015 Innate immunity response to selected cyanobacterial peptides Z. Moosova 1,∗ , O. Adamovsky 1 , M. Pekarova 2 , L. Svihalkova Sindlerova 2 , L. Kubala 2 , L. Blaha 2 1 Masaryk University, Research Centre for Toxic Compounds in the Environment (RECETOX), Brno, Czech Republic 2 Academy of Sciences, Institute of Biophysics, Brno, Czech Republic
Cyanobacteria are ubiquitous photosynthesizing gram-negative bacteria frequently associated with toxic water bloom. Cyanobacterial production of a wide range of hepatotoxins, neurotoxins and dermatotoxins affects water quality, which is a growing environmental and health concern worldwide. Hepatotoxic microcystins (MCs) are very stable in the environment, and due to high risk to human health the WHO has established a provisional guideline of 1 g/L for MC-LR in drinking water. Up to now, only limited set of known cyanobacterial toxins were evaluated for immunomodulatory potencies. In our study, we have focused on characterization of the immunomodulation of MCs and aeruginosin in two experimental designs: (i) direct effects of cyanobacterial peptides on murine macrophage (RAW 264.7 cell line) viability and inflammatory response, (ii) and modulation of macrophage responses by cyanobacterial toxins in co-exposures with model stimuli (lipopolysaccharides, LPS). Cells were exposed to different concentrations of three MC congeners (1–1000 nM) and aeruginosin-865 (0.1–50 M) for 24 h. The production of nitric oxide (NO) in cell supernatants was quantified by Griess reagent method, cell viability was monitored using neutral red uptake assay and BCA protein assay kit, and the levels of pro-inflammatory cytokines (such as interleukin IL-6 and tumor necrosis factor TNF␣) were detected by ELISA method. Our results show that observed effects are congenerspecific. MC-LR was able to significantly affect the activation of macrophages, however, other most common MCs (-RR, -YR) and aeruginosin-865 did not alter immune responds. MC-LR also promoted production of NO and cytokines in co-exposure with LPS used as an established positive control. These results suggest that MC-LR may have significant immunomodulatory activities in very low environmentally relevant concentrations. The research was
S223
supported by the grant from the Czech National Science Foundation ˇ GP13-27695P). (GACR http://dx.doi.org/10.1016/j.toxlet.2015.08.659
P09-016 Genetic polymorphisms in the cytokine genes and risk of type 2 diabetes and its complications in a group of Turkish population A. Karakaya 1,∗ , I. Ates 1 , A. Güvenc¸ 2 , D. Altuner 3 1 Ankara University, Faculty of Pharmacy, Department of Toxicology, Ankara, Turkey 2 Kırsehir City Hospital, Kırsehir, Turkey 3 Erzincan University, Faculty of Medicine, Pharmacology, Erzincan, Turkey
Aim: Type 2 diabetes (Diabetes Mellitus, DM) is a hyperglycaemic metabolic disease related with carbohydrate, protein and lipid metabolism dysfunction due to the decrease of insulin secretion. Several genetic and environmental factors are playing major role in the development of the diabetic complications. Relationship between the inflammation generation and diabetic complications has been showed in recent studies. Following the formation of the oxidative stress after the disorder of the lipid metabolism, the levels of the reactive oxygen species (ROS) increase and insulin resistance develops consequently. Cytokines are important in regulation of the homeostatic mechanisms such as inflammation and tissue repair. Thus, variations in their levels and structures are the reasons for the occurrence of various diseases. The single nucleotide polymorphisms (SNP) forming on the cytokine genes increase the risk of disease development. Recent studies showed the relationship between some inflammatory cytokine gene polymorphisms and the development of complication in patients with diabetes. Based on this, our aim was searching and the evaluating the possible relations between the TNF-␣ (-308), IL-1 (+3953), IL-6 (-174) gene polymorphisms and the development of the complications in a Type 2 diabetic Turkish patient population. Methods: The study was performed by using PCR-RFLP method. Results and conclusions: In regards to our data, both TNF-␣ and IL-1 gene polymorphisms are significantly related with the development of both disease (3,27 fold and 2,15 fold respectively) and complications (4,15 fold and 2,11 fold respectively). On the other hand, there is a little but not significant effect of IL-6 gene polymorphism on the disease occurrence and complication development. http://dx.doi.org/10.1016/j.toxlet.2015.08.660
P09-017 Assessment of sensitization and photosensitization of avobenzone based on DPRA C. Benevenuto ∗ , L. Gaspar FCFRP-USP, Pharmaceutical Sciences, RIBEIRÃO PRETO, Brazil Purpose: Contact allergies are considered complex diseases it reaches 15–20% of the population. Among the cosmetic allergens, UV-filters have been considered emerging allergens and avobenzone (AVO) is one of the most used UVA-filter. The commonly methods to assess skin sensitization still use animals and among the few non-animal methods is the DPRA (direct peptide reactivity assay), which considers that if a substance can react with skin