کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
8417359 | 1545683 | 2015 | 10 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
A robust and scalable TCR-based reporter cell assay to measure HIV-1 Nef-mediated T cell immune evasion
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کلمات کلیدی
NFATCTLTCrHLA class I - HLA کلاس Inuclear factor of activated T-cells - عامل هسته سلول های فعال شده فعال استImmune evasion - فرار از ایمنcytotoxic T lymphocyte - لنفوسیت T سیتوتوکسیکNef - نافHIV - ویروس نقص ایمنی انسانی human immunodeficiency virus - ویروس نقص ایمنی انسانیT cell receptor - گیرنده سلول TT-cell receptor - گیرنده لنفوسیت T
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
بیوتکنولوژی یا زیستفناوری
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چکیده انگلیسی
HIV-1 evades cytotoxic T cell responses through Nef-mediated downregulation of HLA class I molecules from the infected cell surface. Methods to quantify the impact of Nef on T cell recognition typically employ patient-derived T cell clones; however, these assays are limited by the cost and effort required to isolate and maintain primary cell lines. The variable activity of different T cell clones and the limited number of cells generated by re-stimulation can also hinder assay reproducibility and scalability. Here, we describe a heterologous T cell receptor reporter assay and use it to study immune evasion by Nef. Induction of NFAT-driven luciferase following co-culture with peptide-pulsed or virus-infected target cells serves as a rapid, quantitative and antigen-specific measure of T cell recognition of its cognate peptide/HLA complex. We demonstrate that Nef-mediated downregulation of HLA on target cells correlates inversely with T cell receptor-dependent luminescent signal generated by effector cells. This method provides a robust, flexible and scalable platform that is suitable for studies to measure Nef function in the context of different viral peptide/HLA antigens, to assess the function of patient-derived Nef alleles, or to screen small molecule libraries to identify novel Nef inhibitors.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Immunological Methods - Volume 426, November 2015, Pages 104-113
Journal: Journal of Immunological Methods - Volume 426, November 2015, Pages 104-113
نویسندگان
Gursev Anmole, Xiaomei T. Kuang, Mako Toyoda, Eric Martin, Aniqa Shahid, Anh Q. Le, Tristan Markle, Bemuluyigza Baraki, R. Brad Jones, Mario A. Ostrowski, Takamasa Ueno, Zabrina L. Brumme, Mark A. Brockman,