VH gene usage and CDR3 analysis of B cell receptor in the peripheral blood of patients with PBC

Angela Foreman, Brigitte Lemercier, Annick Lim, Phillipe Kourlisky, Thomas Kenny, M. Eric Gershwin, Marie Lise Gougeon

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Abstract

We have analyzed the IgM, IgG and IgA BCR repertoire in PBC patients by means of quantitative RT-PCR and CDR3-spectratyping with immunoscope technology. PBMC from 35 PBC patients and 18 normal controls were analyzed. Quantitative B cell repertoire analysis of IgM from healthy donors showed the preferential usage of VH3a, VH3b and VH4 families. Very similar VH family usage was observed in IgM B cells from PBC patients. CDR3 spectratyping of IgM BCR rearrangements showed a Gaussian distribution for dominant VH families in control donors, and similar diversity was found for the VH3b family in PBC patients. In contrast, VH3a and VH4 families showed oligoclonal expansions in some patients. Quantitative B cell repertoire analysis of IgG and IgA did not reveal any difference in VH chain distribution in PBC patients as compared to the control donors. Immunoscope profiles of CDR3 length distribution showed several peak expansions in B cells from control donors, particularly for the VH3a and VH4 families. CDR3 length distribution profiles of IgG and IgA from PBC patients were oligoclonal too, with expansions throughout the various VH chains. However, no common expansions within the CDR3 region were found intraindividually between IgG, IgA and IgM, and between patients. In conclusion, immunoscope technology does provide, for the first time, a sensitive and rapid method for detailed immunoglobulin gene usage analysis in peripheral B cells from PBC patients. This study failed to demonstrate preferential B cell rearrangements in the blood of patients with PBC, but this technology may be more successful if applied to the analysis of compartmental B cells (i.e. liver infiltrating B cells).

Original languageEnglish (US)
Pages (from-to)80-86
Number of pages7
JournalAutoimmunity
Volume41
Issue number1
DOIs
StatePublished - Feb 2008

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B-Lymphocytes
Genes
Immunoglobulin M
Immunoglobulin A
Immunoglobulin G
Tissue Donors
Technology
Immunoglobulin Genes
Normal Distribution
Polymerase Chain Reaction
Liver

Keywords

  • Anti-mitochondrial antibodies
  • B cells
  • CDR3 region
  • Primary biliary cirrhosis

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

Cite this

VH gene usage and CDR3 analysis of B cell receptor in the peripheral blood of patients with PBC. / Foreman, Angela; Lemercier, Brigitte; Lim, Annick; Kourlisky, Phillipe; Kenny, Thomas; Gershwin, M. Eric; Gougeon, Marie Lise.

In: Autoimmunity, Vol. 41, No. 1, 02.2008, p. 80-86.

Research output: Contribution to journalArticle

Foreman, A, Lemercier, B, Lim, A, Kourlisky, P, Kenny, T, Gershwin, ME & Gougeon, ML 2008, 'VH gene usage and CDR3 analysis of B cell receptor in the peripheral blood of patients with PBC', Autoimmunity, vol. 41, no. 1, pp. 80-86. https://doi.org/10.1080/08916930701619656
Foreman, Angela ; Lemercier, Brigitte ; Lim, Annick ; Kourlisky, Phillipe ; Kenny, Thomas ; Gershwin, M. Eric ; Gougeon, Marie Lise. / VH gene usage and CDR3 analysis of B cell receptor in the peripheral blood of patients with PBC. In: Autoimmunity. 2008 ; Vol. 41, No. 1. pp. 80-86.
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AB - We have analyzed the IgM, IgG and IgA BCR repertoire in PBC patients by means of quantitative RT-PCR and CDR3-spectratyping with immunoscope technology. PBMC from 35 PBC patients and 18 normal controls were analyzed. Quantitative B cell repertoire analysis of IgM from healthy donors showed the preferential usage of VH3a, VH3b and VH4 families. Very similar VH family usage was observed in IgM B cells from PBC patients. CDR3 spectratyping of IgM BCR rearrangements showed a Gaussian distribution for dominant VH families in control donors, and similar diversity was found for the VH3b family in PBC patients. In contrast, VH3a and VH4 families showed oligoclonal expansions in some patients. Quantitative B cell repertoire analysis of IgG and IgA did not reveal any difference in VH chain distribution in PBC patients as compared to the control donors. Immunoscope profiles of CDR3 length distribution showed several peak expansions in B cells from control donors, particularly for the VH3a and VH4 families. CDR3 length distribution profiles of IgG and IgA from PBC patients were oligoclonal too, with expansions throughout the various VH chains. However, no common expansions within the CDR3 region were found intraindividually between IgG, IgA and IgM, and between patients. In conclusion, immunoscope technology does provide, for the first time, a sensitive and rapid method for detailed immunoglobulin gene usage analysis in peripheral B cells from PBC patients. This study failed to demonstrate preferential B cell rearrangements in the blood of patients with PBC, but this technology may be more successful if applied to the analysis of compartmental B cells (i.e. liver infiltrating B cells).

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