Generation of heterohybridomas secreting human immunoglobulins; pokeweed mitogen prestimulation is highly effective but phytohemagglutinin drives most B cells into apoptosis

2.50
Hdl Handle:
http://hdl.handle.net/2336/32036
Title:
Generation of heterohybridomas secreting human immunoglobulins; pokeweed mitogen prestimulation is highly effective but phytohemagglutinin drives most B cells into apoptosis
Authors:
Arinbjarnarson, Sturla; Valdimarsson, Helgi
Citation:
J. Immunol. Methods 2002, 259(1-2):139-48
Issue Date:
1-Jan-2002
Abstract:
Human monoclonal antibodies have commonly been generated by forming hybridomas of stable lymphoblastoid cell lines and Epstein-Barr virus (EBV)-transformed human B cells that have been exposed to phytohaemagglutin (PHA)-stimulated T cells. However, this technique has predominantly given rise to IgM- but very rarely IgG- or IgA-producing clones. We now report that, regardless of prior EBV infection, pokeweed mitogen (PWM) stimulation of human peripheral blood mononuclear cells (PBMCs) generated much higher numbers of IgM-, IgA- and IgG-producing B cells than did stimulation with PHA. Fusion of PWM-stimulated PBMCs with a mouse myeloma cell line also gave rise to 7- to 12-fold higher numbers of IgG- and IgA-producing heterohybridomas than PBMCs that were prestimulated with PHA. Judged by Annexin V staining, stimulation with PHA induced a very high rate of B cell apoptosis within 24 h, whereas, even after 7 days, PWM stimulation only induced marginal B cell apoptosis. This should explain why PHA is much inferior to PWM in stimulating immunoglobulin (Ig) production in vitro and in generating immunoglobulin-producing human B cell hybridomas. It is concluded that PWM stimulation may greatly facilitate the generation of human monoclonal antibodies of all isotypes.
Description:
To access publisher full text version of this article. Please click on the hyperlink in Additional Links field
Additional Links:
http://www.sciencedirect.com/science/article/B6T2Y-44HYK7Y-F/1/555f2f4b7dfd6724aa36a9e9cc638090

Full metadata record

DC FieldValue Language
dc.contributor.authorArinbjarnarson, Sturla-
dc.contributor.authorValdimarsson, Helgi-
dc.date.accessioned2008-07-15T13:43:34Z-
dc.date.available2008-07-15T13:43:34Z-
dc.date.issued2002-01-01-
dc.date.submitted2008-07-15-
dc.identifier.citationJ. Immunol. Methods 2002, 259(1-2):139-48en
dc.identifier.issn0022-1759-
dc.identifier.pmid11730849-
dc.identifier.doi10.1016/S0022-1759(01)00504-X-
dc.identifier.urihttp://hdl.handle.net/2336/32036-
dc.descriptionTo access publisher full text version of this article. Please click on the hyperlink in Additional Links fielden
dc.description.abstractHuman monoclonal antibodies have commonly been generated by forming hybridomas of stable lymphoblastoid cell lines and Epstein-Barr virus (EBV)-transformed human B cells that have been exposed to phytohaemagglutin (PHA)-stimulated T cells. However, this technique has predominantly given rise to IgM- but very rarely IgG- or IgA-producing clones. We now report that, regardless of prior EBV infection, pokeweed mitogen (PWM) stimulation of human peripheral blood mononuclear cells (PBMCs) generated much higher numbers of IgM-, IgA- and IgG-producing B cells than did stimulation with PHA. Fusion of PWM-stimulated PBMCs with a mouse myeloma cell line also gave rise to 7- to 12-fold higher numbers of IgG- and IgA-producing heterohybridomas than PBMCs that were prestimulated with PHA. Judged by Annexin V staining, stimulation with PHA induced a very high rate of B cell apoptosis within 24 h, whereas, even after 7 days, PWM stimulation only induced marginal B cell apoptosis. This should explain why PHA is much inferior to PWM in stimulating immunoglobulin (Ig) production in vitro and in generating immunoglobulin-producing human B cell hybridomas. It is concluded that PWM stimulation may greatly facilitate the generation of human monoclonal antibodies of all isotypes.en
dc.language.isoenen
dc.publisherNorth-Holland Pub. Co.en
dc.relation.urlhttp://www.sciencedirect.com/science/article/B6T2Y-44HYK7Y-F/1/555f2f4b7dfd6724aa36a9e9cc638090en
dc.subject.meshAnimalsen
dc.subject.meshApoptosisen
dc.subject.meshB-Lymphocytesen
dc.subject.meshCell Line, Transformeden
dc.subject.meshHumansen
dc.subject.meshHybridomasen
dc.subject.meshImmunoglobulin Isotypesen
dc.subject.meshMiceen
dc.subject.meshPhytohemagglutininsen
dc.subject.meshPokeweed Mitogensen
dc.titleGeneration of heterohybridomas secreting human immunoglobulins; pokeweed mitogen prestimulation is highly effective but phytohemagglutinin drives most B cells into apoptosisen
dc.typeArticleen
dc.contributor.departmentDepartment of Immunology, National University Hospital, 101 Reykjavík, Iceland.en
dc.identifier.journalJournal of immunological methodsen
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