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Transgenic human lambda 5 rescues the murine lambda 5 nullizygous phenotype.

M E Donohoe1, G B Beck-Engeser, N Lonberg

  • 1Department of Microbiology, University of Miami School of Medicine, Miami, FL 33101, USA.

Journal of Immunology (Baltimore, Md. : 1950)
|May 9, 2000
PubMed
Summary
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The human lambda 5 (hu lambda 5) gene functions similarly to its mouse counterpart, showing stage-specific expression in developing B cells. This study demonstrates hu lambda 5 can substitute for mouse lambda 5 (m lambda 5) in vivo, aiding B cell development.

Area of Science:

  • Immunology
  • Molecular Biology
  • Developmental Biology

Background:

  • The pre-B cell receptor is crucial for B lymphocyte development, composed of Ig mu H chain, VpreB, Ig alpha, Ig beta, and lambda 5.
  • The human lambda 5 (hu lambda 5) gene is homologous to the murine lambda 5 (m lambda 5) gene and is active in pro-B and pre-B lymphocytes.

Purpose of the Study:

  • To investigate the transcriptional regulation of the hu lambda 5 gene in pro-B and pre-B lymphocytes in vivo.
  • To determine if the hu lambda 5 polypeptide can functionally substitute for the m lambda 5 polypeptide in B cell development.

Main Methods:

  • Generation of transgenic mouse lines carrying a 28-kb genomic fragment of the hu lambda 5 gene.
  • Analysis of hu lambda 5 gene expression at mRNA and protein levels using flow cytometry and immunoprecipitation.

Related Experiment Videos

  • Breeding transgenic hu lambda 5 mice onto an m lambda 5 knockout background to assess functional rescue.

    • Main Results:

    • Transgenic hu lambda 5 expression was detected at high levels in bone marrow pro-B and pre-B cells, indicating the presence of necessary regulatory elements within the 28-kb fragment.
    • The hu lambda 5 polypeptide was found on the cell surface and associated with mouse Ig mu and VpreB.
    • The hu lambda 5 transgene rescued the pre-B lymphocyte block in m lambda 5-/- mice.

    Conclusions:

    • The hu lambda 5 polypeptide can biochemically and functionally substitute for m lambda 5 in vivo during pre-B lymphocyte differentiation and proliferation.
    • The transgenic hu lambda 5 model provides a system to study molecular requirements for B cell development.