Anti-CD40 plus interleukin-4-activated human naive B cell lines express unmutated immunoglobulin genes with intraclonal heavy chain isotype variability

European Journal of Immunology
L GalibertS Lebecque

Abstract

Combination of anti-CD40 antibody and interleukin-4 (IL-4) induces B cell clonal expansion reminiscent of the T-dependent proliferation following antigenic challenge in vivo. We have analyzed the usage of CH genes and the presence or absence of somatic mutations within the progeny of a single human naive B cell activated with anti-CD40 + IL-4. To address this issue, single-cell cultures of naive (sIgD+) tonsillar B lymphocytes expressing the VH1-restricted G8 idiotype were set up. After culture and RNA extraction, VH1+ Ig mRNA were reverse-transcribed, amplified by polymerase chain reaction and sequenced. A single sIgD+ B cell could generate clones expressing mu, gamma 1, gamma 3, or epsilon, illustrating that the progeny of a single cell can express different isotypes in response to the same stimulus in vitro. The rate of somatic mutations affecting the immunoglobulin variable heavy chain gene was indistinguishable from the background of errors introduced by Taq polymerase.

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Citations

Jan 5, 2002·International Journal of Cancer. Journal International Du Cancer·Nongnit Laytragoon-LewinHåkan Mellstedt
Jan 15, 2011·Archivum Immunologiae Et Therapiae Experimentalis·Sonia NéronJean-François Leblanc
Apr 12, 2000·Journal of Immunological Methods·N R Ling
Jan 1, 1997·Immunity·S DenépouxS Lebecque
Jul 1, 1997·European Journal of Immunology·I Fugier-VivierY J Liu
Dec 21, 2000·The Journal of Immunology : Official Journal of the American Association of Immunologists·G F Widhopf, T J Kipps
Oct 22, 2003·The Journal of Immunology : Official Journal of the American Association of Immunologists·Jessie F Fecteau, Sonia Néron
Jan 24, 2008·PloS One·Martina WiesnerAndreas Moosmann

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