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Influenza Virus H5N1 infection can induce ROS production
Researchers at Huazhong Agricultural University in Wuhan, China have established that infection by the avian influenza virus H5N1 induces reactive oxygen species (ROS) production, eventually leading to viral replication and host cell death. In conducting their study, the researchers' goals were to investigate the pathogenesis of H5N1 in regards to oxidative stress, and to propose utilizing the expression of endogenous genes with antioxidant effects, such as Cu/Zn superoxide dismutase (SOD1), as a potential treatment.
To elucidate the various impacts H5N1 generated in ROS levels of infected cells, as well as the extent of antioxidant effects of overexpressed SOD1, the researchers decided to evaluate and compare several conditions using the human alveolar epithelial cell line A549 infected with H5N1 strain DW, and transfected with plasmid vector PCAGGS with and without the human SOD1 insert. Since mitochondrial changes are closely related to the overall oxidative state of the cell, the researchers decided to include how mitochondrial function was affected in their study. To assess mitochondrial activity, the researchers observed ATP production, and mitochondrial membrane potential (MMP) and ROS, through the use of an ATP assay kit, JC-10 dye, and a superoxide indicator, respectively. With the results, the researchers could conclude that healthy mitochondrial function was heavily impeded by H5N1 infection, and that SOD1 overexpression could attenuate these detrimental effects in A549 cells.
In the descriptions of the materials and methods used to determine these conclusions, it is notable that the cultured A549 cells were able to be incubated in JC-10 at 15 M. Other alternatives for JC-10, such as JC-1, have been known to precipitate at concentrations as low as 1 M. JC-10's high aqueous solubility enables it to be used at higher concentrations, which in turn improves cell loading and helps generate a stronger fluorescent signal. Ultimately, JC-10 is an ideal MMP indicator for use in studies with live cells.
To elucidate the various impacts H5N1 generated in ROS levels of infected cells, as well as the extent of antioxidant effects of overexpressed SOD1, the researchers decided to evaluate and compare several conditions using the human alveolar epithelial cell line A549 infected with H5N1 strain DW, and transfected with plasmid vector PCAGGS with and without the human SOD1 insert. Since mitochondrial changes are closely related to the overall oxidative state of the cell, the researchers decided to include how mitochondrial function was affected in their study. To assess mitochondrial activity, the researchers observed ATP production, and mitochondrial membrane potential (MMP) and ROS, through the use of an ATP assay kit, JC-10 dye, and a superoxide indicator, respectively. With the results, the researchers could conclude that healthy mitochondrial function was heavily impeded by H5N1 infection, and that SOD1 overexpression could attenuate these detrimental effects in A549 cells.
In the descriptions of the materials and methods used to determine these conclusions, it is notable that the cultured A549 cells were able to be incubated in JC-10 at 15 M. Other alternatives for JC-10, such as JC-1, have been known to precipitate at concentrations as low as 1 M. JC-10's high aqueous solubility enables it to be used at higher concentrations, which in turn improves cell loading and helps generate a stronger fluorescent signal. Ultimately, JC-10 is an ideal MMP indicator for use in studies with live cells.
References
- Xian Lin, Ruifang Wang, Wei Zou, Xin Sun, Xiaokun Liu, Lianzhong Zhao, Shengyu Wang and Meilin Jin. The Influenza Virus H5N1 Infection Can Induce ROS Production for Viral Replication and Host Cell Death in A549 Cells Modulated by Human Cu/Zn Superoxide Dismutase (SOD1) Overexpression. Viruses 2016, 8(1), 13; doi:10.3390/v8010013
- JC-10. AAT Bioquest, n.d. Web. 27 June 2016
Original created on October 6, 2016, last updated on October 6, 2016
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