Assessment of iNKT phenotypes that distinguishes treatment responders from nonresponders in acute GVHD patients, and GVHD onset from healthy immune reconstitution in allogeneic HSCT patients. (A) Representative visualization of tSNE (viSNE) plot from patients after HSCT (n = 11) and patients at GVHD diagnosis that later were corticosteroid (CS) responsive (n = 1) or refractory (n = 3). Each column shows the marker expression of a user-defined channel range. The cell number has been normalized to include the same number of cells from each group. (B) Summary of HLA-II expression in iNKT cells (n = 18) at GVHD diagnosis. The patients were retrospectively stratified by corticosteroid response after GVHD diagnosis. (C) Frequency of CXCR3 cells within the CD161HLA-II population in patients at GVHD diagnosis who were retrospectively stratified by corticosteroid response. (D) Phenotypic expression of iNKT cells from 34 patients on day 30 after HSCT with myeloablative conditioning (MAC). The patients analyzed did not display any clinical manifestations of GVHD. After day 30, 10 of the patients would develop GVHD (red) but 24 would not (blue). Tx, Transplantation. Means ± standard error of the mean (B,D). *P = .05-.01. ns, not significant.