In vitro induction of somatic mutations in human peripheral B lymphocytes.

  • Authors:
    • A Zafiropoulos
    • M Kandilogiannaki
    • C Dahlenborg
    • C A Borrebaeck
    • E Krambovitis
  • View Affiliations

  • Published online on: October 1, 2000     https://doi.org/10.3892/ijmm.6.4.475
  • Pages: 475-559
Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )
Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )


Abstract

The major cellular and molecular factors that lead to the induction of somatic mutations in human B cells remain unclear. This study describes an approach which allowed us to single-cell culture antigen-specific B cells after in vitro immunization. Using single strand conformational polymorphism analysis on the descendant cultures we were able to demonstrate that the Ig gene of B cells was induced to mutate. Three clones were isolated from a single cell culture of naive B cells immunized against the semi-conserved region of V3 of HIV-1, which contained two point mutations at the CDR2 region leading to amino acid codon change. Two clones were also obtained from a single cell culture of memory B cells immunized against the recall antigen tetanus toxoid. These clones showed one point mutation at the CDR1-FR2 border region leading to amino acid codon change. Fab constructs from the mutated culture of naive B cells immunized against V3 were cloned to a procaryotic expression vector. The expression products of two clones showed anti-V3 specificity in ELISA using three different conjugates. Our results suggest that somatic mutations can be induced in human peripheral B cells through specific interaction with antigen and antigen-activated T cells.

Related Articles

Journal Cover

Oct 2000
Volume 6 Issue 4

Print ISSN: 1107-3756
Online ISSN:1791-244X

Sign up for eToc alerts

Recommend to Library

Copy and paste a formatted citation
x
Spandidos Publications style
Zafiropoulos A, Kandilogiannaki M, Dahlenborg C, Borrebaeck C and Krambovitis E: In vitro induction of somatic mutations in human peripheral B lymphocytes.. Int J Mol Med 6: 475-559, 2000
APA
Zafiropoulos, A., Kandilogiannaki, M., Dahlenborg, C., Borrebaeck, C., & Krambovitis, E. (2000). In vitro induction of somatic mutations in human peripheral B lymphocytes.. International Journal of Molecular Medicine, 6, 475-559. https://doi.org/10.3892/ijmm.6.4.475
MLA
Zafiropoulos, A., Kandilogiannaki, M., Dahlenborg, C., Borrebaeck, C., Krambovitis, E."In vitro induction of somatic mutations in human peripheral B lymphocytes.". International Journal of Molecular Medicine 6.4 (2000): 475-559.
Chicago
Zafiropoulos, A., Kandilogiannaki, M., Dahlenborg, C., Borrebaeck, C., Krambovitis, E."In vitro induction of somatic mutations in human peripheral B lymphocytes.". International Journal of Molecular Medicine 6, no. 4 (2000): 475-559. https://doi.org/10.3892/ijmm.6.4.475