Abstract
A detailed methodology is described for the successful immortalization of B lymphocytes to give lymphoblastoid cell lines using Epstein-Barr (EB) virus. A virus stock is prepared from B95-8 or Akata cell line, which spontaneously release high titers of virus, and the immortalizing titer of the preparation determined. B lymphocytes from fresh or cryopreserved samples are separated from the whole cell population, infected with EB virus and cultured for 4 weeks for the establishment of a lymphoblastoid cell line. The presence of EB virus encoded proteins in the immortalized cell line can be detected by indirect immunofluorescent staining.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Bird AG, McLachlan SM, Britton S (1981). Cyclosporin A promotes spontaneous outgrowth in vitro of Epstein-Barr virus-induced B cell lines. Nature 289: 300–301.
Boyum A (1968). Separation of leucocytes from blood and bone marrow. Scand J Clin Lab Invest 21 (suppl. 97): 38–89.
Henle G, Henle W, Diehl W (1968). Relation of Burkitt's tumor-associated herpes-type virus to infectious mononucleosis. Proc Natl Acad Sci USA 59: 94–101.
Luka J, Kallin B, Klein G (1979). Induction of the Epstein-Barr virus (EBV) cycle in latently infected cells by n-butyrate. Virology 94: 228–231.
Miller G, Shope T, Lisco, H, Stitt D, Lipman M (1972). Epstein-Barr virus: Transformation, cytopathic changes and viral antigens in squirrel monkey and marmoset leukocytes. Proc Natl Acad Sci USA 69: 383–387.
Nadler LM, Stashenko P, Hardy R, van Agthoven A, Terhorst C, Schlossman SF (1981). Characterisation of a human B cell specific antigen (B2) distinct from B1. J Immunol 126: 1941–1943.
Nilsson K, Klein G, Henle W, Henle G (1971). The establishment of lymphoblastoid cell lines from adult and from foetal human lymphoid tissue and its dependence on EBV. Int J Cancer 8: 443–450.
Pattengale PL, Smith RW, Gerber P (1973). Selective transformation of B lymphocytes by EB virus. Lancet (ii): 93–94.
Pope JH, Horne MK, Scott W (1968). Transformation of foetal human leucocytes in vitro by filtrates of a human leukaemic cell line containing herpes-like virus. Int J Cancer 3: 857–866.
Rickinson AB, Moss DJ, Pope JH (1979). Long-term T cell-mediated immunity to Epstein-Barr virus in man, II: Components necessary for regression in virusinfected leukocyte cultures. Int J Cancer 23: 610–617.
Reedman BM, Klein G (1973). Cellular localisation of an Epstein-Barr virus (EBV) associated complement-fixing antigen in producer and non-producer lymphoblastoid cell lines. Int J Cancer 11: 499–520.
Rosen A, Gergely P, Jondal M, Klein G, Britton S (1977). Polyclonal Ig production after Epstein-Barr virus infection of human lymphocytes in vitro. Nature 267: 52–54.
Sudgen B, Phelps M, Domoradzki J (1979). The DNA of Epstein-Barr virus is amplified in transformed lymphocytes. J Virol 31: 590–595.
Takada K (1984). Cross-linking of cell surface immunoglobulin induces Epstein-Barr virus in Burkitt lymphoma lines. Int J Cancer 33: 27–32.
Yao QY, Rickinson AB, Epstein MA (1985). A reexamination of the Epstein-Barr virus carrier state in healthy seropositive individuals. Int J Cancer 35: 35–42.
zur Hausen H, O'Neill F, Freese UK (1978). Persisting oncogenic herpesvirus induced by the tumour promoter TPA. Nature 272: 373–375.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Lam, K.M.C., Crawford, D.H. Method for generation of human B lymphoblastoid cell lines using Epstein-Barr virus. Methods Cell Sci 17, 67–74 (1995). https://doi.org/10.1007/BF00986653
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00986653