American Society of Hematology

Figure 4

$Figure 4. Analysis of EBV genome load in IgM memory B cells isolated from XLP blood. (A) PBMCs from 2 XLP patients, XLP5 and XLP6, were stained with mAbs specific for CD20, IgM, IgD and CD27 to identify naive (CD20+IgM+IgD+ CD27−), nonswitched memory (CD20+IgM+IgD+CD27+) and class-switched memory (CD20+IgM−IgD−CD27+) B-cell populations; numbers indicate the percentage distribution of CD20+ B cells between the 3 populations (left panel). Naive and nonswitched memory cells were subsequently isolated by FACS sorting and the isolated populations reanalyzed, giving percentage purities as shown (center and right panels). (B) The histogram shows the results of quantitative PCR analysis to determine the EBV load in unfractionated CD20+ B cells (), naive B cells () and nonswitched memory B cells (■) isolated from the 2 donors shown in panel A above. Data (mean and SD of 3 replicates) are reported as EBV genome copies per 106 cells.$

Analysis of EBV genome load in IgM memory B cells isolated from XLP blood. (A) PBMCs from 2 XLP patients, XLP5 and XLP6, were stained with mAbs specific for CD20, IgM, IgD and CD27 to identify naive (CD20⁺IgM⁺IgD⁺ CD27⁻), nonswitched memory (CD20⁺IgM⁺IgD⁺CD27⁺) and class-switched memory (CD20⁺IgM⁻IgD⁻CD27⁺) B-cell populations; numbers indicate the percentage distribution of CD20⁺ B cells between the 3 populations (left panel). Naive and nonswitched memory cells were subsequently isolated by FACS sorting and the isolated populations reanalyzed, giving percentage purities as shown (center and right panels). (B) The histogram shows the results of quantitative PCR analysis to determine the EBV load in unfractionated CD20⁺ B cells (), naive B cells () and nonswitched memory B cells (■) isolated from the 2 donors shown in panel A above. Data (mean and SD of 3 replicates) are reported as EBV genome copies per 10⁶ cells.

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