About: Establishment of Epstein-Barr Virus Growth-transformed Lymphoblastoid Cell Lines

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About: Establishment of Epstein-Barr Virus Growth-transformed Lymphoblastoid Cell Lines Goto Sponge NotDistinct Permalink

An Entity of Type : schema:ScholarlyArticle, within Data Space : wasabi.inria.fr associated with source document(s)

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type	Academic Article research paper schema:ScholarlyArticle
isDefinedBy	Covid-on-the-Web dataset
title	Establishment of Epstein-Barr Virus Growth-transformed Lymphoblastoid Cell Lines
Creator	Bhaduri-Mcintosh, S Hill, E Hui-Yuen, J Koganti, S Mcallister, S
source	Medline; PMC
abstract	Infection of B cells with Epstein-Barr virus (EBV) leads to proliferation and subsequent immortalization, resulting in establishment of lymphoblastoid cell lines (LCL) in vitro. Since LCL are latently infected with EBV, they provide a model system to investigate EBV latency and virus-driven B cell proliferation and tumorigenesis(1). LCL have been used to present antigens in a variety of immunologic assays(2, 3). In addition, LCL can be used to generate human monoclonal antibodies(4, 5) and provide a potentially unlimited source when access to primary biologic materials is limited(6, 7). A variety of methods have been described to generate LCL. Earlier methods have included the use of mitogens such as phytohemagglutinin, lipopolysaccharide(8), and pokeweed mitogen(9) to increase the efficiency of EBV-mediated immortalization. More recently, others have used immunosuppressive agents such as cyclosporin A to inhibit T cell-mediated killing of infected B cells(7, 10-12). The considerable length of time from EBV infection to establishment of cell lines drives the requirement for quicker and more reliable methods for EBV-driven B cell growth transformation. Using a combination of high titer EBV and an immunosuppressive agent, we are able to consistently infect, transform, and generate LCL from B cells in peripheral blood. This method uses a small amount of peripheral blood mononuclear cells that are infected in vitroclusters of cells can be demonstrated. The presence of CD23 with EBV in the presence of FK506, a T cell immunosuppressant. Traditionally, outgrowth of proliferating B cells is monitored by visualization of microscopic clusters of cells about a week after infection with EBV. Clumps of LCL can be seen by the naked eye after several weeks. We describe an assay to determine early if EBV-mediated growth transformation is successful even before microscopic clusters of cells can be demonstrated. The presence of CD23(hi)CD58(+) cells observed as early as three days post-infection indicates a successful outcome.
has issue date	2011-11-08(xsd:dateTime)
bibo:doi	10.3791/3321
bibo:pmid	22090023
has license	cc-by-nc-nd
sha1sum (hex)	3c919c2857ddedf0a444aa214761e16e4676d50b
schema:url	https://doi.org/10.3791/3321
resource representing a document's title	Establishment of Epstein-Barr Virus Growth-transformed Lymphoblastoid Cell Lines
has PubMed Central identifier	PMC3308597
has PubMed identifier	22090023
schema:publication	J Vis Exp
resource representing a document's body	covid:3c919c2857ddedf0a444aa214761e16e4676d50b#body_text
is schema:about of	named entity 'ESTABLISHMENT' named entity 'VIRUS' named entity 'B CELLS' named entity 'B CELL PROLIFERATION' named entity 'TO INVESTIGATE' named entity 'leads' named entity 'Cell' named entity 'TRANSFORMED' named entity 'LYMPHOBLASTOID CELL' named entity 'MODEL SYSTEM' named entity 'LEADS' named entity 'TUMORIGENESIS' named entity 'ESTABLISHMENT' named entity 'EPSTEIN-BARR VIRUS' named entity 'IMMORTALIZATION' named entity 'RESULTING IN' named entity 'LATENCY' named entity 'CELL LINES' named entity 'IN VITRO' named entity 'SUBSEQUENT' named entity 'EPSTEIN-BARR VIRUS' named entity 'EBV' named entity 'DRIVEN' named entity 'LYMPHOBLASTOID CELL' named entity 'PROLIFERATION' named entity 'GROWTH' named entity 'INFECTION' named entity 'LCL' named entity 'INFECTED' named entity 'PROVIDE' named entity 'CELL LINES' named entity 'EBV' named entity 'proliferation' named entity 'investigate' named entity 'cell lines' named entity 'Lymphoblastoid' named entity 'Virus' named entity 'EBV' named entity 'lymphoblastoid cell' named entity 'FK506' named entity 'lipopolysaccharide' named entity 'supernatant' named entity 'fluorescence' named entity 'live cells' named entity 'CD58' named entity 'cryopreservation' named entity 'clusters of cells' named entity 'liquid nitrogen' named entity 'antibodies' named entity 'phytohemagglutinin' named entity 'hemocytometer' named entity 'supernatant' named entity 'titer' named entity 'EBV infection' named entity 'light microscopy' named entity 'infection' named entity 'Trypan blue' named entity 'CD19' named entity 'EBV' named entity '37°C' named entity 'antibody' named entity '37°C' named entity 'clusters of cells' named entity 'CD58' named entity 'EBV' named entity 'cell growth' named entity 'mitogens' named entity 'CD19' named entity 'centrifuge' named entity 'EBV' named entity 'Cy5' named entity 'buffy coat' named entity 'light microscopy' named entity 'culture medium' named entity 'lymphocyte' named entity 'EBV infection' named entity 'EBV'

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