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Blood, Vol. 111, Issue 3, 1344-1356, February 1, 2008
Clonal expansion of immunoglobulin M+CD27+ B cells in HCV-associated mixed cryoglobulinemia
Blood Charles et al.
111: 1344
Supplemental materials for: Charles et al
Files in this Data Supplement:
- Figure S1. No loss of activation markers from cryopreserved cells (PDF, 68.1 KB) -
PBMCs from an HCV patient were stained for CD3, CD20 and the indicated activation/differentiation markers either immediately after isolation (pink solid histograms) or following cryopreservation and thawing (blue line). Flow cytometric analysis of CD20+ gated lymphocytes (A). Healthy donor PBMCs were stained either immediately after isolation (red), immediately after 72 hours of activation with 3 µg/ml PHA (green), or after identical activation plus cryopreservation (blue). Flow cytometric analysis of CD20+ gated lymphocytes (B).
- Figure S2. Flow cytometric analysis of B cells. B cells were identified by gating on lymphocytes, followed by exclusion of dead cells and selection of CD19+ CD20+ cells (PDF, 54 KB) -
(A). Percentages of B cells among groups, and among HCV+ subjects with clonal or polyclonal Ig (B). Percentages of CD5+ B cells (C). Graphs are of gated B cells. Horizontal bars indicate medians. P values were calculated by the Kruskal-Wallis test.
- Figure S3. HCV+ patients with clonal Ig have expanded IgM+κ+CD21lowCD27+ B cells (PDF, 110 KB) -
Increased  / light chain ratios in HCV+ subjects with clonal Ig compared to an HCV Ab− subject and an HCV+ patient with polyclonal Ig (A). HCV+ individuals with clonal Ig have increased +CD27+ B cells (B). CD27+ B cells in HCV+ patients with clonal Ig are predominantly IgM+ (C). IgM+ B cells in HCV+ patients with clonal Ig are predominantly IgDlow/neg (D). A large proportion of CD27+ B cells in certain HCV+ patients with clonal Ig are CD21low (E). The graphs in the right column depict selected CD19+, CD20+ B cell subsets among all patients. Horizontal bars indicate medians. P values were calculated by the Kruskal-Wallis test, and if P < 0.05, the Dunn multiple comparison test was used as a posttest to compare the HCV+MC+ with HCV+MC− groups.
- Figure S4. The expanded IgM+κ+CD27+ B cells from selected HCV+ patients with clonal Ig are predominantly IgDlow/neg (PDF, 43.7 KB) -
Analysis was performed on gated +CD27+ (A) and IgM+CD27+ (B) B cells.
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