کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
10889350 | 1080828 | 2008 | 10 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Engineering higher affinity T cell receptors using a T cell display system
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کلمات کلیدی
PBSmAbpeptide-MHCβ2MCDRPEPOVATCrBSA - BSAβ2 microglobulin - β2 میکروگلوبولینbovine serum albumin - آلبومین سرم گاوMonoclonal antibody - آنتی بادی مونوکلونالRetroviral vectors - بردارهای Retroviralconstant - ثابتPhosphate buffered saline - فسفات بافر شورVariable - متغیرMHC - مجموعه سازگاری بافتی اصلیmajor histocompatibility complex - مجموعه سازگاری بافتی اصلیcomplementarity determining region - منطقه تعریف مکملProtein engineering - مهندسی پروتئینAffinity - وابستگیPeptide - پپتید T cell receptor - گیرنده سلول TT cell receptors - گیرنده های T سلول
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
بیوتکنولوژی یا زیستفناوری
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چکیده انگلیسی
The T cell receptor (TCR) determines the cellular response to antigens, which are presented on the surface of target cells in the form of a peptide bound to a product of the major histocompatibility complex (pepMHC). The response of the T cell depends on the affinity of the TCR for the pepMHC, yet many TCRs have been shown to be of low affinity, and some naturally occurring T cell responses are poor due to low affinities. Accordingly, engineering the TCR for increased affinity for pepMHC, particularly tumor-associated antigens, has become an increasingly desirable goal, especially with the advent of adoptive T cell therapies. For largely technical reasons, to date there have been only a handful of TCRs engineered in vitro for higher affinity using well established methods of protein engineering. Here we report the use of a T cell display system, using a retroviral vector, for generating a high-affinity TCR from the mouse T cell clone 2C. The method relies on the display of the TCR, in its normal, signaling competent state, as a CD3 complex on the T cell surface. A library in the CDR3α of the 2C TCR was generated in the MSCV retroviral vector and transduced into a TCR-negative hybridoma. Selection of a high-affinity, CD8-independent TCR was accomplished after only two rounds of flow cytometric sorting using the pepMHC SIYRYYGL/Kb (SIY/Kb). The selected TCR contained a sequence motif in the CDR3α with characteristics of several other TCRs previously selected by yeast display. In addition, it was possible to directly use the selected T cell hybridoma in functional assays without the need for sub-cloning, revealing that the selected TCR was capable of mediating CD8-independent activity. The method may be useful in the direct isolation and characterization of TCRs that could be used in therapies with adoptive transferred T cells.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Immunological Methods - Volume 339, Issue 2, 31 December 2008, Pages 175-184
Journal: Journal of Immunological Methods - Volume 339, Issue 2, 31 December 2008, Pages 175-184
نویسندگان
Adam S. Chervin, David H. Aggen, John M. Raseman, David M. Kranz,