A virus-neutralising antibody is not cytotoxic in vitro

Andreas Kolb, C Bruce A Whitelaw

    Research output: Contribution to journalArticle

    1 Citation (Scopus)

    Abstract

    Hybridoma cell lines are characterized by a preferential loss of the heavy chain gene. This observation has led to the theory that the immunoglobulin heavy chain possesses an intrinsic cytotoxic activity in some cell types. We have generated transgenic mice expressing the heavy and light chain genes of the virus-neutralising antibody A1 carrying constant domains of the human gamma1 and kappa isotype. Heavy chain and light chain transgenes were under trancriptional control of identical promoter regions derived from the mammary gland specific ovine beta-lactoglobulin gene. The copy number of the heavy chain transgene was consistently lower than the copy number of the light chain gene in all lines of transgenic mice. Moreover, the light chain gene was expressed in significant excess of the heavy chain gene in the lactating mammary gland in all transgenic lines. In several transgenic lines, the differences in antibody expression were greater than could be explained by the differences in transgene copy number. One potential cause of this phenotype could be a cytotoxic effect of free heavy chain protein in embryonic cells (resulting in differences in copy number) or mammary epithelial cells (resulting in differences in transgene expression). We therefore directly assessed the effect of the expression of free A1 heavy chain protein in epithelial cell lines and in murine embryonic stem cells. However, full-length A1 heavy chain mRNA and protein could be expressed transiently and stably in both epithelial and embryonic stem cells and had no detectable effect on cell viability. Taken together, these findings argue against an inherent cytotoxicity of the free A1 heavy chain protein in epithelial or embryonic cells.
    Original languageEnglish
    Pages (from-to)677-689
    Number of pages13
    JournalMolecular Immunology
    Volume43
    Issue number6
    Early online date24 May 2005
    DOIs
    Publication statusPublished - 1 Feb 2006

    Fingerprint

    Neutralizing Antibodies
    Viruses
    Transgenes
    Genes
    Light
    Human Mammary Glands
    Embryonic Stem Cells
    Transgenic Mice
    Proteins
    Epithelial Cells
    Cell Line
    Lactoglobulins
    Immunoglobulin Heavy Chains
    Hybridomas
    Genetic Promoter Regions
    In Vitro Techniques
    Cell Survival
    Sheep
    Breast
    Phenotype

    Keywords

    • Animals
    • Antibodies, Viral
    • Cell Survival
    • Embryo, Mammalian
    • Epithelial Cells
    • Gene Dosage
    • Gene Expression Regulation
    • Humans
    • Immunoglobulin Heavy Chains
    • Immunoglobulin Light Chains
    • Mice
    • Mice, Transgenic
    • Recombinant Fusion Proteins
    • Stem Cells
    • Transgenes
    • Antibody
    • Cytotoxicity
    • RMCE
    • Cre recombinase
    • Mammary gland
    • Transgenic mice

    Cite this

    A virus-neutralising antibody is not cytotoxic in vitro. / Kolb, Andreas; Whitelaw, C Bruce A.

    In: Molecular Immunology, Vol. 43, No. 6, 01.02.2006, p. 677-689.

    Research output: Contribution to journalArticle

    Kolb, Andreas ; Whitelaw, C Bruce A. / A virus-neutralising antibody is not cytotoxic in vitro. In: Molecular Immunology. 2006 ; Vol. 43, No. 6. pp. 677-689.
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