Abstract
Epstein–Barr virus (EBV) is a herpes virus which in vitro efficiently immortalizes nearly all human B lymphocytes. The lymphoblastoid diploid cell lines (LCL’s) thus generated preserve the characteristics of the cells initially infected by the virus: the cells produce and secrete immunoglobulins and also express these molecules on their surface. A selection of specific antibody-producing cells (i.e., antigen-committed cells) before EBV-infection or when LCL’s have already been established, enables isolation of monoclonal cell lines that secrete specific antibodies. If selection of antigen-committed cells is not feasible, secretion of specific antibodies by cloned LCL’s in limiting dilution cultures enables isolation of the desired cell lines. The method allows the production of human IgM, IgG, IgA, and IgE monoclonal antibodies from any individual. Monoclonal antibodies produced by the EBV method resemble the antibody repertoire of the donor of the lymphocytes. Human monoclonal antibodies are promising reagents for passive immunization.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Nilsson K, Ponten J (1975) Classification and biological nature of established human hematopoietic cell lines. Int J Cancer 15:321–341
Steinitz M, Klein G, Koskimies S, Mäkelä O (1977) EB virus-induced B lymphocyte cell lines producing specific antibody. Nature 269:420–422
Kuppers R (2003) B cells under influence: transformation of B cells by Epstein-Barr virus. Nat Rev Immunol 3:801–812
Nemerow GR, Wolfert R, McNaughton ME, Cooper NR (1985) Identification and characterization of the Epstein-Barr virus receptor on human lymphocytes-B and its relationship to the C3d complement receptor (Cr-2). J Virol 55:347–351
Heath E, Begue-Pastor N, Chaganti S, Croom-Carter D, Shannon-Lowe C, Kube D, Feederle R, Delecluse HJ, Rickinson AB, Bell AI (2012) Epstein-Barr virus infection of naive B cells in vitro frequently selects clones with mutated immunoglobulin genotypes: implications for virus biology. PLoS Pathog 8(5):e1002697
Ehlin-Henriksson B, Gordon J, Klein G (2003) B-lymphocyte subpopulations are equally susceptible to Epstein-Barr virus infection, irrespective of immunoglobulin isotype expression. Immunology 108:427–430
Brown NA, Miller G (1982) Immunoglobulin expression by human lymphocytes-B clonally transformed by Epstein-Barr virus. J Immunol 128:24–29
Miyawaki T, Kubagawa H, Butler JL, Cooper MD (1988) Ig isotypes produced by Ebv-transformed B-cells as a function of age and tissue distribution. J Immunol 140:3887–3892
Stein LD, Chan MA, Hibi T, Dosch HM (1986) Epstein-Barr virus-induced Ige production in limiting dilution cultures of normal human B-cells. Eur J Immunol 16:1167–1170
Steinitz M, Klein G (1980) Ebv-transformation of surface Iga-positive human-lymphocytes. J Immunol 125:194–196
Shannon-Lowe C, Baldwin G, Feederle R, Bell A, Rickinson A, Delecluse HJ (2005) Epstein-Barr virus-induced B-cell transformation: quantitating events from virus binding to cell outgrowth. J Gen Virol 86:3009–3019
Gil Y, Levy-Nabot S, Steinitz M, Laskov R (2007) Somatic mutations and activation-induced cytidine deaminase (AID) expression in established rheumatoid factor-producing lymphoblastoid cell line. Mol Immunol 44:494–505
Chezar I, Lobel-Lavi L, Steinitz M, Laskov R (2008) Ongoing somatic hypermutation of the rearranged VH but not of the V-lambda gene in EBV-transformed rheumatoid factor-producing lymphoblastoid cell line. Mol Immunol 46:80–90
Laskov R, Yahud V, Hamo R, Steinitz M (2011) Preferential targeting of somatic hypermutation to hotspot motifs and hypermutable sites and generation of mutational clusters in the IgVH alleles of a rheumatoid factor producing lymphoblastoid cell line. Mol Immunol 48:733–745
Miller G, Lipman M (1973) Comparison of yield of infectious virus from clones of human and simian lymphoblastoid lines transformed by Epstein-Barr-virus. J Exp Med 138:1398–1412
Steinitz M, Tamir S, Frodin JE, Lefvert AK, Mellstedt H (1988) Human monoclonal anti-idiotypic antibodies.1. Establishment of immortalized cell-lines from a tumor patient treated with mouse monoclonal-antibodies. J Immunol 141:3516–3522
Traggiai E, Becker S, Subbarao K, Kolesnikova L, Uematsu Y, Gismondo MR, Murphy BR, Rappuoli R, Lanzavecchia A (2004) An efficient method to make human monoclonal antibodies from memory B cells: potent neutralization of SARS coronavirus. Nat Med 10:871–875
Acknowledgment
The help of Dr. A. Mahler in preparing the manuscript is greatly appreciated.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2014 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Steinitz, M. (2014). Production of Human Monoclonal Antibodies by the Epstein–Barr Virus Method. In: Steinitz, M. (eds) Human Monoclonal Antibodies. Methods in Molecular Biology, vol 1060. Humana, Totowa, NJ. https://doi.org/10.1007/978-1-62703-586-6_6
Download citation
DOI: https://doi.org/10.1007/978-1-62703-586-6_6
Published:
Publisher Name: Humana, Totowa, NJ
Print ISBN: 978-1-62703-585-9
Online ISBN: 978-1-62703-586-6
eBook Packages: Springer Protocols