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Fig. 1.

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Fig. 1. Lymphocyte subset analysis of the peripheral blood (Case 1) by flow cytometry using Primary Immunodeficiency Orientation Tube (PIDOT) panel. Patient’s B-cell subsets show strong reduction in CD27 expression including unswitched memory B cells/plasma cells and switched memory B cells. Impaired maturation of post-GC B-cells is a hallmark of common variable immunodeficiency (CVID). (A) After gating lymphocytes on CD45 channel, forward scatter channel (FSC) and side scatter channel (SSC), CD3 and CD19 in combination with TCRγδ were used to define subsets of lymphocytes; T-cells (upper left), B-cells (lower right) and NK cells (lower left). (B) CD16+CD56+ and CD3 markers were used to define NK cells (upper left). (C) The CD3+ T-cells were divided into CD4+ (lower right), CD8+ (upper left) and double negative T-cells (DNT) (lower left). (D) The double negative T-cells were subdivided into two subpopulations according to the expression of TCRγδ; TCRγδ- T-cells (upper left) and TCRγδ+ T-cells (upper right). (E) CD4+ T-cell subsets were subdivided into naïve (CD27+CD45RA+; upper right), central memory/transitional memory (CM/TM; CD27+CD45RA-; upper left), effector memory (EM; CD27-CD45RA-; lower left) and terminally differentiated (TD; CD27-CD45RA+; lower right) CD4+ T-cells. (F) CD8+ T-cell subsets were subdivided into naïve (CD27+CD45RA+; upper right), CM/TM (CD27+CD45RA-; upper left), EM (CD27-CD45RA-; lower left) and TD (CD27-CD45RA+; lower right) CD8+ T-cells. For CD8+ T-cells, one additional subpopulation is defined, effector CD27dim (middle right). (G) Subdivision of the B-cell subset demonstrated pre-germinal center B-cells (Pre-GC; CD27-CD16-CD56-; lower left) and reduced post-germinal center B-cells (Post-GC; CD27+CD16-CD56-; upper left). (H) Pre-GC B-cells were sub-classified into mature naïve B-cells (CD27-smIgM+IgD++; upper right; brown) and transitional/immature B-cells (CD27-smIgM++IgD+; lower right; magenta) by the intensity of surface membrane immunoglobulin (smIg) M and D markers. (I) Likewise, post-GC B-cells were sub-classified into two different subsets based on the expression level of smIgM and smIgD; unswitched memory B cells/plasma cells (MBC/PC; CD27+smIgM+IgD+; upper right) and switched MBC/PC (CD27+smIgM-IgD-; lower left). In this plot, proportion of post-GC cells was generally reduced.
Lab Med Online 2023;13:53~59
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