کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
8417189 | 1545677 | 2016 | 35 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Flow cytometric gating for spleen monocyte and DC subsets: differences in autoimmune NOD mice and with acute inflammation
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کلمات کلیدی
(SSC)(DCS)(Rt)multicolor flow cytometryC57BL/6 - C57BL / 6Antigen presenting cells - آنتیژن ارائه سلولinflammation - التهاب( توروم) interferon - اینترفرونLineage - خط راستRoom temperature - دمای اتاقNon-obese diabetic - دیابتی غیر چاقیconventional dendritic cells - سلول های دندریتی معمولیDendritic cells - سلول های دندریتیکMonocyte-derived dendritic cells - سلولهای دندریتیک مونوسیتplasmacytoid dendritic cells - سلولهای دندریتیک پلاسماسیتیوئیدRegulatory T cells - سلولهای تی تنظیمکنندهfluorescence minus one - فلورسانس منهای یکnon-obese diabetic mice - موش های دیابتی غیر چاقmean fluorescence intensity - میانگین شدت فلورسانسImmune homeostasis - هوموستاز ایمنparaformaldehyde - پارافرمالدهیدforward scatter - پراکندگی رو به جلوside scatter - پراکندگی سمت
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
بیوتکنولوژی یا زیستفناوری
پیش نمایش صفحه اول مقاله
چکیده انگلیسی
The role of antigen presenting cells (APCs) in the pathogenesis of autoimmune and other inflammatory diseases is now better understood due to advances in multicolor flow cytometry, gene expression analysis of APC populations, and functional correlation of mouse to human APC populations. A simple but informative nomenclature of conventional and plasmacytoid dendritic cell subsets (cDC1, cDC2, pDC) and monocyte-derived populations incorporates these advances, but accurate subset identification is critical. Ambiguous gating schemes and alterations of cell surface markers in inflammatory condition can make comparing results between studies difficult. Both acute inflammation, such as TLR-ligand stimulation, and chronic inflammation as found in mouse models of autoimmunity can alter DC subset gating. Here, we address these issues using in vivo CpG stimulation as an example of acute inflammation and the non-obese diabetic (NOD) mouse as a model of chronic inflammation. We provide a flow cytometric antibody panel and gating scheme that differentiate 2 monocytic and 3 DC subsets in the spleen both at steady state and after CpG stimulation. Using this method, we observed differences in the composition of NOD DCs that have been previously reported, and newly identified increases in the number of NOD monocyte-derived DCs. Finally, we established a protocol for DC phosphoflow to measure the phosphorylation state of intracellular proteins, and use it to confirm functional differences in the identified subsets. Therefore, we present optimized methods for distinguishing monocytic and DC populations with and without inflammation and/or autoimmunity associated with NOD mice.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Immunological Methods - Volume 432, May 2016, Pages 4-12
Journal: Journal of Immunological Methods - Volume 432, May 2016, Pages 4-12
نویسندگان
Matthew B. Dong, M. Jubayer Rahman, Kristin V. Tarbell,