Fig. 3.
Fig. 3. Three-color immunofluorescence analysis of different stages of B-lineage cells in BM. / BM cells from an 11-month-old healthy infant and an XLA patient (P6) were fixed, permeabilized, and then stained with PE-labeled anti-VpreB and PC5-labeled anti-CD19 mAbs in combination with an FITC-labeled antihuman μH chain Ab. B-lineage cells in BM lymphoid cells were gated by a combination of VpreB and CD19 expression, as described in Figure 1. (A) According to the degree of VpreB and μH chain expression, BM B-lineage cells were arbitrarily divided into 5 fractions: μ−SL++ (R1), μlowSL++ (R2), μlowSL+ (R3), μlowSL− (R4), and μhighSL− (R5) cells. R1, R2, R3, R4, and R5 fractions comprise 7.6%, 9.9%, 19.0%, 38.8%, and 24.6%, respectively, of the normal BM or 76.8%, 14.8%, 3.9%, 3.1%, and 1.0% of the XLA BM. (B) The cell size of each fraction of BM-lineage cells was assessed by forward scatter. In normal BM, large cells were more enriched in the R2 fraction. By contrast, all fractions were mainly composed of small cells in XLA BM.

Three-color immunofluorescence analysis of different stages of B-lineage cells in BM.

BM cells from an 11-month-old healthy infant and an XLA patient (P6) were fixed, permeabilized, and then stained with PE-labeled anti-VpreB and PC5-labeled anti-CD19 mAbs in combination with an FITC-labeled antihuman μH chain Ab. B-lineage cells in BM lymphoid cells were gated by a combination of VpreB and CD19 expression, as described in Figure 1. (A) According to the degree of VpreB and μH chain expression, BM B-lineage cells were arbitrarily divided into 5 fractions: μSL++ (R1), μlowSL++ (R2), μlowSL+ (R3), μlowSL(R4), and μhighSL (R5) cells. R1, R2, R3, R4, and R5 fractions comprise 7.6%, 9.9%, 19.0%, 38.8%, and 24.6%, respectively, of the normal BM or 76.8%, 14.8%, 3.9%, 3.1%, and 1.0% of the XLA BM. (B) The cell size of each fraction of BM-lineage cells was assessed by forward scatter. In normal BM, large cells were more enriched in the R2 fraction. By contrast, all fractions were mainly composed of small cells in XLA BM.

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