title: Characterization of antigen-specific B cells using nominal antigen-coated flow-beads.
abstract: In order to characterize the reactivity of B cells against nominal antigens, a method based on the coupling of antigens onto the surface of fluorescent core polystyrene beads was developed. We first demonstrate that murine B cells with a human MOG-specific BCR are able to interact with MOG-coated beads and do not recognize beads coated with human albumin or pp65. B cells purified from human healthy volunteer blood or immunized individuals were tested for their ability to interact with various nominal antigens, including viral, vaccine, self and alloantigens, chosen for their usefulness in studying a variety of pathological processes. A substantial amount of B cells binding self-antigen MOG-coated beads can be detected in normal blood. Furthermore, greater frequencies of B cell against anti-Tetanic Toxin or anti-EBNA1 were observed in primed individuals. This method can reveal increased frequencies of anti-HLA committed B cells in patients with circulating anti-HLA antibodies compared to unsensitized patients and normal individuals. Of interest, those specific CD19 cells were preferentially identified within CD27(-)IgD(+) (i-e naïve) subset. These observations suggest that a broad range of medical situations could benefit from a tool that allows the detection, the quantification and the characterization of antigen-specific blood B cells.
authors.affiliation: INSERM, UMR 1064, Nantes, France ; CHU de Nantes, ITUN, Nantes, France ; Université de Nantes, Faculté de Médecine, Nantes, France.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Degauque
authors.forename: Annie
authors.initials: A
authors.lastname: Elong  Ngono
authors.affiliation: INSERM, UMR 1064, Nantes, France ; CHU de Nantes, ITUN, Nantes, France ; Université de Nantes, Faculté de Médecine, Nantes, France.
authors.forename: Annie Elong
authors.initials: AE
authors.lastname: Ngono
authors.affiliation: INSERM, UMR 1064, Nantes, France ; CHU de Nantes, ITUN, Nantes, France ; Université de Nantes, Faculté de Médecine, Nantes, France.
authors.forename: Ahmed
authors.initials: A
authors.lastname: Akl
authors.affiliation: INSERM, UMR 1064, Nantes, France ; CHU de Nantes, ITUN, Nantes, France ; Université de Nantes, Faculté de Médecine, Nantes, France.
authors.forename: Maud
authors.initials: M
authors.lastname: Lepetit
authors.affiliation: INSERM, UMR 1064, Nantes, France ; CHU de Nantes, ITUN, Nantes, France ; Université de Nantes, Faculté de Médecine, Nantes, France.
authors.forename: Romain
authors.initials: R
authors.lastname: Crochette
authors.affiliation: INSERM, UMR 1064, Nantes, France ; CHU de Nantes, ITUN, Nantes, France ; Université de Nantes, Faculté de Médecine, Nantes, France.
authors.forename: Magali
authors.initials: M
authors.lastname: Giral
authors.affiliation: INSERM, UMR 1064, Nantes, France ; CHU de Nantes, ITUN, Nantes, France ; Université de Nantes, Faculté de Médecine, Nantes, France.
authors.forename: Julie
authors.initials: J
authors.lastname: Lepourry
authors.affiliation: INSERM, UMR 1064, Nantes, France ; CHU de Nantes, ITUN, Nantes, France ; Université de Nantes, Faculté de Médecine, Nantes, France.
authors.forename: Annaick
authors.initials: A
authors.lastname: Pallier
authors.affiliation: INSERM, UMR 1064, Nantes, France ; CHU de Nantes, ITUN, Nantes, France ; Université de Nantes, Faculté de Médecine, Nantes, France.
authors.forename: Stéphanie
authors.initials: S
authors.lastname: Castagnet
authors.affiliation: INSERM, UMR 1064, Nantes, France ; CHU de Nantes, ITUN, Nantes, France ; Université de Nantes, Faculté de Médecine, Nantes, France.
authors.forename: Emilie
authors.initials: E
authors.lastname: Dugast
authors.affiliation: CHU de Nantes, ITUN, Nantes, France.
authors.forename: Cécile
authors.initials: C
authors.lastname: Guillot-Gueguen
authors.affiliation: INSERM, CIC 004, Nantes, France.
authors.forename: Marylène
authors.initials: M
authors.lastname: Jacq-Foucher
authors.affiliation: Université de Nantes, Faculté de Médecine, Nantes, France ; INSERM, UMR892, Nantes, France.
authors.forename: Xavier
authors.initials: X
authors.lastname: Saulquin
authors.affiliation: Etablissement Français du Sang, Laboratoire HLA, Nantes, France.
authors.forename: Anne
authors.initials: A
authors.lastname: Cesbron
authors.affiliation: INSERM, UMR 1064, Nantes, France ; CHU de Nantes, ITUN, Nantes, France ; Université de Nantes, Faculté de Médecine, Nantes, France ; INSERM, CIC 004, Nantes, France.
authors.forename: David
authors.initials: D
authors.lastname: Laplaud
authors.affiliation: INSERM, UMR 1064, Nantes, France ; CHU de Nantes, ITUN, Nantes, France ; Université de Nantes, Faculté de Médecine, Nantes, France.
authors.forename: Arnaud
authors.initials: A
authors.lastname: Nicot
authors.affiliation: INSERM, UMR 1064, Nantes, France ; CHU de Nantes, ITUN, Nantes, France ; Université de Nantes, Faculté de Médecine, Nantes, France.
authors.forename: Sophie
authors.initials: S
authors.lastname: Brouard
authors.affiliation: INSERM, UMR 1064, Nantes, France ; CHU de Nantes, ITUN, Nantes, France ; Université de Nantes, Faculté de Médecine, Nantes, France.
authors.forename: Jean-Paul
authors.initials: JP
authors.lastname: Soulillou
mesh_terms: D000328:Adult; D000368:Aged; D000818:Animals; D000941:Antigens / Q000737:chemistry* / Q000276:immunology*; D018941:Antigens, CD19 / Q000378:metabolism; D001324:Autoantigens / Q000276:immunology; D001402:B-Lymphocytes / Q000166:cytology* / Q000276:immunology* / Q000378:metabolism; D005260:Female; D005434:Flow Cytometry / Q000379:methods*; D005456:Fluorescent Dyes / Q000737:chemistry; D006680:HLA Antigens / Q000276:immunology; D006801:Humans; D007114:Immunization; D016030:Kidney Transplantation; D008297:Male; D051379:Mice; D008863:Microspheres; D008875:Middle Aged; D011108:Polymers / Q000737:chemistry; D014612:Vaccines / Q000276:immunology; D014764:Viral Proteins / Q000276:immunology; D055815:Young Adult
chemical_list: D000941:Antigens; D018941:Antigens, CD19; D001324:Autoantigens; D005456:Fluorescent Dyes; D006680:HLA Antigens; D011108:Polymers; D014612:Vaccines; D014764:Viral Proteins; C004216:bioplex