OXIDATIVE STRESS IN LYMPHOBLASTOID CELL LINES FROM AUTISTIC CHILDREN
Abstract
Objective. In this study, we evaluated the in vitro susceptibility of autistic children to various oxidative stressors as compared with to unaffected, age-matched controls. Background. Numerous studies have suggested oxidative stress plays a role in the pathogenesis of autism. Oxidative stress results from an imbalance between the production of reactive oxygen species (ROS) and a decrease in either the efficiency of the endogenous antioxidant defense mechanisms or the ability to effectually scavenge free radicals. We propose that there is a direct link between oxidative stress and cell death in children with autism, with deficient glutathione levels as the underlying mechanism. Methods. Lymphoblastoid cell lines (LCLs) from affected and control children from the Autism Genetic Research Exchange (AGRE) and Coriell Cell Repository were treated with the proapoptotic agent, DMNQ, for different times. Cell viability, cell death, apoptosis, and necrosis rates were analyzed in these treated cell lines, both at baseline and in the presence of oxidative stressors, using flow cytometry. In addition, the formation of ROS was quantified using fluorescence. Results. Preliminary results have shown increased levels of ROS in those LCLs from children with autism as compared to controls at baseline conditions. Significantly higher apoptotic rates were found in the control LCLs (p<0.05) at almost all time points, whereas the autistic LCLs had higher proportion of cell death (p<0.1). Conclusion. Overall, the results from this study will provide a better understanding of the underlying molecular mechanisms surrounding the pathogenesis of autism particularly looking at mechanisms of programmed cell death in this condition
OXIDATIVE STRESS IN LYMPHOBLASTOID CELL LINES FROM AUTISTIC CHILDREN
Melnick Auditorium
Objective. In this study, we evaluated the in vitro susceptibility of autistic children to various oxidative stressors as compared with to unaffected, age-matched controls. Background. Numerous studies have suggested oxidative stress plays a role in the pathogenesis of autism. Oxidative stress results from an imbalance between the production of reactive oxygen species (ROS) and a decrease in either the efficiency of the endogenous antioxidant defense mechanisms or the ability to effectually scavenge free radicals. We propose that there is a direct link between oxidative stress and cell death in children with autism, with deficient glutathione levels as the underlying mechanism. Methods. Lymphoblastoid cell lines (LCLs) from affected and control children from the Autism Genetic Research Exchange (AGRE) and Coriell Cell Repository were treated with the proapoptotic agent, DMNQ, for different times. Cell viability, cell death, apoptosis, and necrosis rates were analyzed in these treated cell lines, both at baseline and in the presence of oxidative stressors, using flow cytometry. In addition, the formation of ROS was quantified using fluorescence. Results. Preliminary results have shown increased levels of ROS in those LCLs from children with autism as compared to controls at baseline conditions. Significantly higher apoptotic rates were found in the control LCLs (p<0.05) at almost all time points, whereas the autistic LCLs had higher proportion of cell death (p<0.1). Conclusion. Overall, the results from this study will provide a better understanding of the underlying molecular mechanisms surrounding the pathogenesis of autism particularly looking at mechanisms of programmed cell death in this condition