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Blood, Vol. 113, Issue 3, 604-611, January 15, 2009
Functional Epstein-Barr virus reservoir in plasma cells derived from infected peripheral blood memory B cells
Blood Al Tabaa et al.
113: 604
Supplemental materials for: Al Tabaa et al
Files in this Data Supplement:
- Figure S1. Characterization of EBV memory B-cell reservoir in blood compartment (JPG, 77.9 KB)
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(A) Isolation of CD19+ B cells from blood samples. (B) Quantification of EBV-DNA and enumeration of BZLF1- and gp350-secreting cells after memory B cells polyclonal activation.
- Figure S2. Detection of BZLF1 or gp350 products in B95-8 cells (JPG, 18.1 KB)
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BZLF1-and gp350 expressions were explored in five experiments by ELISpot and intracytoplasmic immunofluorescence assays.
- Figure S3. EBV-antigens production in acyclovir-treated B95-8 cells (JPG, 32.9 KB)
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Cells were untreated (black square) or treated for 15 days with acyclovir at concentration of 100 µM (white square). (A) B95-8 cells were tested for BZLF1 and gp350 expression by immunofluorescence assay. (B) Cells were tested by ELISpot assays for the secretion of BZLF1 and gp350 products.
- Figure S4. Reproducibility for EBV-infected B cells producing BLF1 or gp350 (JPG, 66.6 KB)
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Highly purified peripheral blood B cells from 13 healthy EBV carriers were stimulated and cultured for 5 days. BZLF1- or gp350-secreting cells were enumerated by ELISpot assays. The Figure illustrated results of two different experiments.
- Figure S5. Sequential activation of enriched B-cell subsets (JPG, 57.9 KB)
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Memory B-cell (A), IgM memory B-cell (B) and naïve B-cell (C) subsets were polyclonally stimulated and analyzed by flow cytometry based on CD38 expression. The phenotype was checked over the culture period for each B-cell subset. Monoparametric fluorescence histograms were showed at day 0, day 3, and day 6 that included the cell incubation step of the ELISpot assays.
- Figure S6. Differentiation of enriched B-cell subsets (JPG, 90.3 KB)
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Memory B-cell (A), IgM memory B-cell (B) and naïve B-cell (C) subsets were polyclonally stimulated and analyzed by flow cytometry based on CD20 and CD138 expression over the culture period. Histogram dot plots were showed at day 0 and day 6 that included the cell incubation step of the ELISpot assays.
- Figure S7. Reproducibility for EBV-infected B-cell subsets producing BLF1 or gp350 (JPG, 62.2 KB)
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BZLF1- or gp350-secreting cells were enumerated in memory B-cell (square) and IgM memory B-cell (circle) subsets by ELISpot assays. The Figure illustrated results of two different experiments. ND, not detected.
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