CD19 signalling improves the Epstein-Barr virus-induced immortalization of human B cell.
Epstein-Barr virus (EBV) infection in vitro immortalizes primary B cells and generates B lymphoblastoid cell lines (LCLs). These EBV-LCLs have been used for several purposes in immunological and genetic studies, but some trials involving these transformations fail for unknown reasons, and several EBV-LCLs do not grow in normal culture. In this study, we improved the immortalization method by CD19 and B-cell receptor (BCR) co-ligation. This method shortens the time required for the immortalization and generation of EBV-LCLs but does not alter the cell phenotype of the LCLs nor the expression of the EBV genes. In particular, the CD19 and BCR co-ligation method was found to be the most effective method examined. EBV-infected B cells induced by CD19 and/or BCR ligation expressed the intracellular latent membrane protein LMP-1 earlier than EBV-infected B cells, and the expression of intracellular LMP-1 was found to be closely related to the time of immortalization. These results suggest that the modified method, using CD19 and/or BCR ligation, may efficiently generate EBV-LCLs, by expressing intracellular LMP-1 at an early stage.
Hur
D Y
DY
Department of Anatomy and Research Center for Immune Modulation, Inje University College of Medicine, 614-735 Busan, South Korea.
Lee
M H
MH
Kim
J W
JW
Kim
J-H
JH
Shin
Y K
YK
Rho
J K
JK
Kwack
K B
KB
Lee
W J
WJ
Han
B G
BG
D018941:Antigens, CD19 / Q000096:biosynthesis*; D001402:B-Lymphocytes / Q000166:cytology* / Q000378:metabolism; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D002469:Cell Separation; D002472:Cell Transformation, Viral; D019309:Epstein-Barr Virus Nuclear Antigens / Q000378:metabolism; D005434:Flow Cytometry; D004854:Herpesvirus 4, Human / Q000378:metabolism*; D006801:Humans; D016130:Immunophenotyping; D007963:Leukocytes, Mononuclear / Q000378:metabolism; D008214:Lymphocytes / Q000821:virology; D010641:Phenotype; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction; D013997:Time Factors; D014763:Viral Matrix Proteins / Q000378:metabolism; D014764:Viral Proteins
D018941:Antigens, CD19; C402641:EBNA-2 protein, Human herpesvirus 4; C037662:EBV-associated membrane antigen, Epstein-Barr virus; D019309:Epstein-Barr Virus Nuclear Antigens; D014763:Viral Matrix Proteins; D014764:Viral Proteins; C417029:EBV-encoded nuclear antigen 1