Fluorescence-activated cell sorting analysis of peripheral blood mononuclear cells reveal a defect in differentiation of type 2 lymphocytes in HSCT-treated GC/JAK3 patients. (A) Unsupervised Uniform Manifold Approximation and Projection (UMAP)²⁷ of single live CD45⁺Lin^–CD3^–CD4^–CD7⁺ cells was applied for CD56, CD16, CD94, NKp46, CD94, CD127, CD161, CD25, CD117, and CRTH2 fluorescence parameters. (B) Supervised analysis of circulating ILCs including two CD56⁺ subsets of NK cells and two CD127⁺ subsets denoted as ILC2 (CRTh2+) and ILCP (ILC precursors, CRTh2^–CD117⁺CD45RA⁺NKp44^–) (manually gating strategy presented in supplemental Figure 1A). (C) UMAP analysis on CD4⁺ T cells, including CXCR3, CCR4, CRTH2, CCR6, CXCR5, CD25, CD127, and CD45RA fluorescence parameters (supplemental Figure 2A). (D) Supervised analysis of circulating T-cell populations. The different subsets were identified as follow: naive (CD45RA⁺), Th1 (Treg^–CD45RA^–CXCR5^–CCR6^–CXCR3⁺CCR4^–), Th17 (Treg^–CD45RA^–CXCR5^–CCR6⁺), Th2 (Treg^–CD45RA^–CXCR5^–CCR6^–CXCR3^–CCR4⁺CRTH2^+/−), and Treg (CD127loCD25⁺) (manually gating strategy is provided in supplemental Figure 1B). (E) UMAP analysis on CD8⁺ type 2 cytotoxic T-cell (Tc2) subset including CCR4 and CRTH2 fluorescence parameters (supplemental Figure 3). (F) Supervised analysis of circulating CD8⁺ Tc2 subset defined as CD45RA^–CD25^–CD94^–CD56^–CXCR5^–CCR6^–CXCR3^–CCR4⁺CRTH2^+/−. Panels B, D, and F: box plots with median ± minimum to maximum. P values were determined with the Kruskal-Wallis test followed by Dunn’s posttest for multiple group comparisons; *P < .05, **P < .005, ***P < .001. n.s., not significant.