Figure 1. DOCK8 is highly expressed in lymphocyte subsets, absent in DOCK8-deficient patients and restored following HSCT.

(A) PBMCs from healthy donors (n = 3) were stained with Abs against CD3, CD4, CD8, CD20, CD56, CD161, and TCR Vα24, Vβ11, and Vα7.2. The cells were then fixed, permeabilized, and stained with anti-DOCK8 mAb. Expression of intracellular DOCK8 in total T cells (CD3⁺), CD4⁺ T cells (CD3⁺CD4⁺CD8^–), CD8⁺ T cells (CD3⁺CD4^–CD8⁺), B cells (CD20⁺CD3^–), NK cells (CD3^–CD56⁺), NKT cells (CD3⁺TCRVα24⁺Vβ11⁺), and MAIT cells (CD3⁺CD161⁺TCRVα7.2⁺) was then determined. Data represent the average geometric MFI ± SEM of different lymphocyte subsets from 3 unrelated donors labeled with anti-DOCK8 mAb less the MFI of cells labeled with isotype control mAb. (B and C) PBMCs from healthy donors (n = 20) or DOCK8-deficient patients before (n = 4) or following HSCT (pHSCT; n = 15–16) were stained with Abs against CD4, CD8, and CD20 before fixing, permeabilization, and staining for DOCK8. DOCK8 expression was determined in total lymphocytes (B), as well as in CD4⁺ T cells, CD8⁺ T cells, and CD20⁺ B cells (C). The histogram in B depicts DOCK8 expression in total lymphocytes from 1 representative healthy donor, and lymphocytes from the same DOCK8-deficient patient before and after transplant as well as an isotype control. The graph in C represents the mean MFI ± SEM of DOCK8 expression (minus MFI of isotype control mAb). Statistical analysis was performed using unpaired t test with Welch’s correction; **P < 0.01, ****P < 0.001.