Human CD20+CD43+CD27+CD5− B cells generate antibodies to capsular polysaccharides of Streptococcus pneumoniae Bert Verbinnen, PhD, Kris Covens, PhD,

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Presentation transcript:

Human CD20+CD43+CD27+CD5− B cells generate antibodies to capsular polysaccharides of Streptococcus pneumoniae Bert Verbinnen, PhD, Kris Covens, PhD, Leen Moens, PhD, Isabelle Meyts, MD, PhD, Xavier Bossuyt, MD, PhD Journal of Allergy and Clinical Immunology Volume 130, Issue 1, Pages 272-275 (July 2012) DOI: 10.1016/j.jaci.2012.04.040 Copyright © 2012 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 1 Increase in the percentage and absolute number of B-1 cells after vaccination with Pneumo23. Six healthy volunteers were intramuscularly vaccinated with the polysaccharide-based vaccine Pneumo23, and PBMCs were collected just before and 1 week after vaccination. The cells were immunofluorescently stained for CD45-V500, CD3–phycoerythrin (PE) Cy7, CD20–allophycocyanin (APC)–eFluor780, CD19-V450, CD43-PE, CD27-APC, CD70–fluorescein isothiocyanate (FITC), and CD5-PerCP-Cy5.5 and were then evaluated by means of flow cytometry. A, Gating strategy for the analysis of CD5− and CD5+ B-1 cells. Doublets were excluded from analysis based on the forward scatter (FSC)–H and FSC-A profile, and T cells were excluded by gating CD3− cells. B-1 cells were defined by being CD19+, CD20+, CD43+, CD27+, CD70−, CD5+, or CD5−. SSC, Side scatter. B, The percentage of B-1 cells among CD19+CD20+ B cells and the percentage of B-1 cells positive or negative for CD5 are shown before (pre) and after (post) vaccination. C, The absolute number of total B-1 cells and CD5− and CD5+ B-1 cells is shown before (pre) and after (post) vaccination. Journal of Allergy and Clinical Immunology 2012 130, 272-275DOI: (10.1016/j.jaci.2012.04.040) Copyright © 2012 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 2 Polysaccharide (PS)–specific antibody secretion by CD5− and CD5+ subsets of the proposed B-1 cell and by naive and memory B cells after vaccination with Pneumo23. Healthy subjects were vaccinated with Pneumo23, and 1 week later, their CD5+ and CD5− proposed B-1 cells (CD19+CD20+CD27+CD43+CD70−), as well as their naive B cells (CD19+CD20+CD27−CD43−) and memory B cells (CD19+CD20+CD27+CD43−), were analyzed for their capacity to secrete IgA, IgM, and IgG anti–caps-PS antibodies by using ELISpot. Cells were plated at 2 × 104 cells per well and incubated for 12 hours at 37°C. A, Representative ELISpot wells from 1 donor are shown for PS1. B, The number of spots for PS1 and PS4 for each B-cell subset is shown for IgA, IgM, and IgG. >>, Excess of spots; —, not done. Journal of Allergy and Clinical Immunology 2012 130, 272-275DOI: (10.1016/j.jaci.2012.04.040) Copyright © 2012 American Academy of Allergy, Asthma & Immunology Terms and Conditions