|کد مقاله||کد نشریه||سال انتشار||مقاله انگلیسی||ترجمه فارسی||نسخه تمام متن|
|5672878||1593426||2018||6 صفحه PDF||سفارش دهید||دانلود کنید|
- Avian splenocyte fixation with a 1:1 ICB dilution does not significantly impact phenotyping.
- NDV in infected splenocytes can be inactivated using a 1:1 ICB dilution for 45Â min at 23Â Â°C and 4Â Â°C.
- 1Â ÃÂ ICB can inactivate 109.5 EID50 of AF-suspended NDV in 45Â min at 23Â Â°C and 4Â Â°C.
- NDV present in infected HD11 cells can be inactivated using 1Â ÃÂ ICB for 45Â min at 4Â Â°C.
Inactivation of Newcastle disease virus (NDV) has been routinely achieved with heat, Î²-propiolactone, binary ethylenimine, ultraviolet light and formalin. However, these strategies have not been tested for cell surface ligand or receptor phenotype in viral-infected chicken immune cells. To study the capacity of fixation buffers to preserve surface markers while inactivating NDV, a primary splenocyte culture was infected with NDV and incubated with a commercial intracellular fixation buffer (ICB), formulated with 4% formaldehyde. Splenocytes were fixed with a 1:2 dilution of ICB in phosphate buffered saline (PBS) for 45Â min at 23Â Â°C or 4Â Â°C and inactivation of NDV was tested in addition to recognition of antigens by antibodies in fixed and non-fixed splenocytes via flow cytometric analysis. The binding and percentage of splenic CD4+ and CD8+ cells were not affected. In addition, NDV titers as high as 109.5 and 107.6 EID50 in allantoic fluid (AF) and macrophages, respectively, were successfully inactivated after 45Â min at 23Â Â°C and 4Â Â°C, confirming the ICB's effectiveness in inactivating high concentrations of NDV. In conclusion, high concentrations of NDV in AF, chicken splenocytes, and macrophages can be inactivated using ICB. Additionally, this method did not compromise cell phenotyping of enriched chicken splenocytes.
Journal: Journal of Virological Methods - Volume 251, January 2018, Pages 1-6