کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
9902251 | 1545796 | 2005 | 13 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Enumeration and characterization of virus-specific B cells by multicolor flow cytometry
دانلود مقاله + سفارش ترجمه
دانلود مقاله ISI انگلیسی
رایگان برای ایرانیان
کلمات کلیدی
BCRASCCFAPR8IFAcomplete Freund's adjuvant - adjuvant دوست کاملincomplete Freund's adjuvant - adjuvant فروند ناقص استantibody secreting cell - آنتی بادی ترشح کننده سلولInfluenza - آنفلوانزا Streptavidin - استرپتاویدینHAU - اینBrdU - بروموداکسی اوریدینhen egg lysozyme - تخم مرغ لیزوزیمhemagglutinin - هماگلوتینینplaque forming units - واحدهای تشکیل پلاکhemagglutinating units - واحدهای هموگلوتینImmune response - پاسخ یا واکنش ایمنیpfu - پفوHEL - کاملB cell receptor - گیرنده سلول B
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
بیوتکنولوژی یا زیستفناوری
پیش نمایش صفحه اول مقاله
چکیده انگلیسی
To better characterize B cell responses induced to influenza virus, we developed an assay to directly quantify and characterize virus-specific B cells. We used purified and biotinylated whole virus as well as the major influenza virus surface antigen, hemagglutinin (HA) to label virus-specific B cells induced by immunization of mice with whole influenza virus in adjuvant. Immunization with adjuvant alone caused non-specific binding of whole virus to a large number of B cells in the draining lymph nodes as assessed by flow cytometry. This precluded the use of whole virus as a specific staining reagent. In contrast, staining with bromelain-cleaved purified and biotinylated influenza virus HA identified a small population of B cells (roughly 1%) only in the draining lymph nodes of virus-immunized mice. FACS-purification and subsequent ELISPOT analysis showed that HA-labeled B cells contained the vast majority of virus-specific antibody-secreting cells at day 10 after immunization. Overall, virus-specific antibody-secreting cells comprised roughly 10% of the HA-labeled cells. Using HA-staining in conjunction with 8-color flow cytometry we further demonstrated that close to 90% of the HA-labeled cells were CD19+ IgDâ CD23â CD24high CD38low germinal center B cells, many of which had incorporated bromodeoxyuridine, indicating recent cell division in vivo. We conclude that viral HA can be used in conjunction with cell surface and intracytoplasmic stains in multicolor flow cytometry to provide detailed phenotypic and functional information on virus HA-specific B cells.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Immunological Methods - Volume 303, Issues 1â2, August 2005, Pages 40-52
Journal: Journal of Immunological Methods - Volume 303, Issues 1â2, August 2005, Pages 40-52
نویسندگان
Virginia P. Doucett, Walter Gerhard, Kristina Owler, Dyan Curry, Lorena Brown, Nicole Baumgarth,