کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
9902268 | 1545797 | 2005 | 13 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Detection of human perforin by ELISpot and ELISA: Ex vivo identification of virus-specific cells
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کلمات کلیدی
streptavidin-alkaline phosphatasePBSGranzymemAbPBMCCTLPHAELISPOTPFNPhytohemagglutininFCSEnzyme-linked immunospot assay - آزمایش ایمنی مرتبط با آنزیمMonoclonal antibody - آنتی بادی مونوکلونالEBV - اپشتین بار ویروسImmunohistochemistry - ایمونوهیستوشیمیIHC - ایمونوهیستوشیمیEnzyme-linked immunosorbent assay - تست الیزاELISA - تست الیزاfetal calf serum - سرم گوساله جنینNK cell - سلول NKPeripheral blood mononuclear cell - سلول تک هسته ای خون محیطیNatural killer cell - سلول قاتل طبیعیcytomegalovirus - سیتومگالوویروسCMV - سیتومگالوویروسCapture ELISA - ضبط الیزاcytotoxic T lymphocyte - لنفوسیت T سیتوتوکسیکPhosphate-buffered saline - محلول نمک فسفات با خاصیت بافریWestern blot - وسترن بلاتEpstein-Barr virus - ویروس Epstein-BarrPerforin - پرفورین
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
بیوتکنولوژی یا زیستفناوری
پیش نمایش صفحه اول مقاله
چکیده انگلیسی
The perforin (PFN) protein is essential for the elimination of target cells by cytotoxic T lymphocytes (CTL) and natural killer (NK) cells. The study of cells releasing PFN has been hampered by a lack of sensitive methods. We therefore produced PFN-reactive monoclonal antibodies (mAb) and developed capture enzyme-linked immunosorbent (ELISA) and enzyme-linked immunospot (ELISpot) assays. Three mAbs were generated and shown to react with unique determinants of PFN. All mAbs recognized intracellular PFN in human peripheral blood mononuclear cell (PBMC) as assessed by flow cytometry and immunohistochemistry. Functional PFN capture ELISA and ELISpot assays were developed utilizing two of the mAbs for capture and the third mAb for detection. When examining PFN release by the YT lymphoma cell line, the ELISpot displayed a greater detection sensitivity than the ELISA. Assessment of PFN release by a CTL clone using ELISpot gave results consistent with a parallel 51Cr-release cytotoxicity assay. Moreover, PFN release by PBMC could be quantified by ELISpot and ELISA after ex vivo stimulation with defined CTL epitopes from common viruses. These novel immunoassays will be valuable for further investigations of the mechanisms underlying granule-mediated apoptosis. In addition, the capture immunoassays could provide tools for studying CTL responses in infectious and tumor diseases as well as for vaccine development.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Immunological Methods - Volume 302, Issues 1â2, July 2005, Pages 13-25
Journal: Journal of Immunological Methods - Volume 302, Issues 1â2, July 2005, Pages 13-25
نویسندگان
Bartek Zuber, Victor Levitsky, Gun Jönsson, Staffan Paulie, Arina Samarina, Susanna Grundström, Sunil Metkar, HÃ¥kan Norell, Glenda G. Callender, Christopher Froelich, Niklas Ahlborg,