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Blood, Vol. 113, Issue 11, 2461-2469, March 12, 2009
Blood-borne human plasma cells in steady state are derived from mucosal immune responses
Blood Mei et al.
113: 2461
Supplemental materials for: Mei et al
Files in this Data Supplement:
- Document 1. Supplemental materials and methods (PDF, 36.6 KB)
- Figure S1. Increased number of circulating plasmablasts/plasma cells, including vaccine-specific cells on day 7 after tetanus/diphtheria vaccination (JPG, 129 KB)
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(A) Detection of icIghigh plasmablasts/plasma cells, including rTT.C-specific plasmablasts in the blood on day 7 after vaccination. PBMCs were stained for cytoplasmatic κ and λ light chains, revealing icIghigh expressing plasmablasts/plasma cells. IcIghigh cells (black histograms) were CD27high, CD38high, CD19+, CD20low, CD138+∕−, CD3− and CD14−, as compared to total PBMCs (grey histograms). Antigen-specific plasmablasts made up median 9% ± 4% SD (range 4.6% to 19.3%) of icIghigh cells in 13 donors analyzed. (B) Gating strategy to identify plasmablasts/plasma cells among PBMCs using CD19, CD27 and CD20 and light scatter parameters. (C) Absolute numbers of circulating CD19+/CD27high/CD20low plasmablasts/plasma cells were calculated in 4 healthy donors before and 7 days after vaccination.
- Figure S2. Detection of human bone marrow plasma cells expressing IgG, IgA or IgM (JPG, 76.5 KB)
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(A–C) Bone marrow plasma cells were identified cytometrically by high expression of CD38. They co-expressed CD138 and contained large amounts of intracellular immunoglobulin. The frequencies of CD38high bone marrow plasma cells among bone marrow mononuclear cells ranged from 0.3 – 2.6% (median 1.1%). (D) Isotype-specific mAbs were used to distinguish icIgA+, icIgG+ and icIgM+ plasma cells among total CD38high bone marrow plasma cells.
- Figure S3. Rare occurrence of increased plasmablast/plasma cell frequency in steady-state (JPG, 69.4 KB)
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Plasmablasts/plasma cells were detected by flow cytometry based on their intracellular expression of immunoglobulin light chains or according to their surface phenotype of CD19+/CD27high/CD20low/CD38high. 3 out of 49 donors analyzed in steady-state displayed a frequency of >0.5% among PBMCs. Both donors analyzed in detail had exceptionally high frequencies of mucosal β7-integrin+ cells among plasmablasts/plasma cells.
- Figure S4. IgA-secreting cells of the human bone marrow express CCR10 and β7-integrin (JPG, 50.1 KB)
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Cytometric analysis of CD38high bone marrow plasma cells co-expressing surface IgA for expression of CCR10 and β7-integrin (open histograms, controls – shaded). A representative data set from 1 out of three donors is shown.
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