کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
9902348 | 1545801 | 2005 | 14 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Expression mapping using a retroviral vector for CD8+ T cell epitopes: Definition of a Mycobacterium tuberculosis peptide presented by H2-Dd
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کلمات کلیدی
PBSmAbCTLHRPELISPOTCD8+ T cells - CD8 + سلول های Tamino acid - آمینو اسیدMonoclonal antibody - آنتی بادی مونوکلونالenzyme-linked immunosorbent spot assay - آنزیم وابسته به ایمنی اسیاب محل آزمونAlgorithm - الگوریتمELISA - تست الیزاEnzyme-linked immunosorbent assay - تست الیزاtransporter associated with antigen processing - حمل کننده همراه با پردازش آنتی ژنreverse transcription - رونویسی معکوسTAP - ضربه زدنCytotoxic T lymphocytes - لنفوسیت های T سیتوتوکسیکmajor histocompatibility complex - مجموعه سازگاری بافتی اصلیMHC - مجموعه سازگاری بافتی اصلیPhosphate-buffered saline - محلول نمک فسفات با خاصیت بافریEpitope mapping - نقشه برداری اپیتوپHorseradish peroxidase - پراکسیداز هوررادیش
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
بیوتکنولوژی یا زیستفناوری
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چکیده انگلیسی
Identification of CD8+ T cell epitopes is important because detection of specific CD8+ T cells after infection or immunization requires prior knowledge of epitope specificity. Furthermore, identification of CD8+ T cell epitopes permits the development of specific preventive and therapeutic approaches to both infections and tumors. Thus far, CD8+ T cell epitopes have been identified either using an overlapping peptide library covering an entire protein, or using algorithms designed to identify likely peptides that bind to major histocompatibility complex (MHC) class I molecules. The synthesis of overlapping peptides can be prohibitively expensive, and the algorithm programs used to predict CD8+ T cell epitopes are not always accurate. Here we describe a retroviral expression system that specifically allows longer polypeptides and shorter peptides to be expressed in the cytoplasm, and thereby to be processed onto class I MHC molecules. T cells from mice that were immunized with a DNA vaccine encoding MPT-51 were probed against MHC-compatible cell lines retrovirally transduced with overlapping gene fragments encoding 120-140 amino acids of the MPT-51 molecule. After further testing of shorter peptide sequences, we identified a CD8+ T cell epitope using cell lines expressing a relatively small number of algorithm-predicted candidate epitopes. We found that one of the requirements for cell surface display of the 20-mer peptide was the need for cotranslational ubiquitination. The restriction molecule was identified as Dd following transduction with MHC class I genes followed by transduction with the oligonucleotide encoding the epitope. The retroviral expression system described here is cost-effective, particularly if the target molecule is large, and could be adapted to identifying T cell epitopes recognized in infectious disease and against tumor cell antigens.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Immunological Methods - Volume 298, Issues 1â2, March 2005, Pages 21-34
Journal: Journal of Immunological Methods - Volume 298, Issues 1â2, March 2005, Pages 21-34
نویسندگان
Taiki Aoshi, Mina Suzuki, Masato Uchijima, Toshi Nagata, Yukio Koide,