Additional file 1 of Post-transplant cyclophosphamide alters immune signatures and leads to impaired T cell reconstitution in allogeneic hematopoietic stem cell transplant
Creators: name: Zhao, Chenchen
name: Bartock, Matthew
name: Jia, Bei
name: Shah, Neal
name: Claxton, David F.
name: Wirk, Baldeep
name: Rakszawski, Kevin L.
name: Nickolich, Myles S.
name: Naik, Seema G.
name: Rybka, Witold B.
name: Ehmann, W Christopher C.
name: Hohl, Raymond J.
name: Valentin, Jessica
name: Bernas-Peterson, Michelle
name: Gerber, Emily M.
name: Zimmerman, Michele
name: Mierski, Joseph A.
name: Mineishi, Shin
name: Zheng, Hong
Publisher: name: figshare
Year: 2022
Subjects: subject: Immunology
subject: FOS: Clinical medicine

description: Additional file 1: Table S1 Patient characteristics. Table S2 Clinical characteristics and outcomes of patients treated with PTCy prophylaxis. Table S3 Conjugated monoclonal antibodies and panel design that used in the flow cytometry analysis. Table S4 Identification of immune cell populations. Fig. S1 Reconstitution of lymphocytes was significantly delayed in patients who received PTCy. Flow-cytometry analysis was performed on PBMCs collected from patient with non-PTCy or PTCy during allo-HSCT. The immune cell components were gated according to defined markers (listed in Table S4). The frequencies of immune cell subsets in PBMCs (A) and their absolute numbers in peripheral blood per μL(B) are exhibited by box-and-whisker plots. Each dot represents the corresponding value from an individual patient. Immune cell subsets that were significantly different between non-PTCy (circle, blue) and PTCy (square, red) groups are shown. Asterisks denote statistically differences comparing the two groups at different timepoints; P values were obtained by the Wilcoxon-rank sum test; *P &lt; 0.05; **P &lt; 0.05; ***P &lt; 0.001; ****P &lt; 0.0001. Fig. S2 Expressions of co-stimulatory molecules and activation markers of T cells under the impact of PTCy after allo-HSCT. The expression of surface inhibitory molecules on CD4+/CD8+ T cells, which are significantly different between the 2 cohorts, is shown through the box-and-whiskers plots. Each dot represents an individual patient. P values were calculated using Wilcoxon rank-sum tests and were corrected for the multiple comparison using the Benjamini–Hochberg adjustment. *P &lt; 0.05; **P &lt; 0.05; ***P &lt; 0.001; ****P &lt; 0.0001. Fig. S3 A Representative flow cytometry data showing IFN-γ, TNF-α and IL-2 expression on CD4+ or CD8+ T cells. B Summarized data of TNF-α and IL-2 expression. P values were calculated using Wilcoxon rank-sum tests and were corrected for multiple comparisons using the Benjamini–Hochberg adjustment. Each dot represents an individual patient. Fig. S4 Comparison of phenotypic markers of T cells on day 30 after allo-HSCT between PTCy recipients according to clinical significance of GVHD. Two groups are defined as no clinically significant GVHD group (ns-GVHD; grade 0–1 aGVHD and mild/moderate cGVHD, n=17) and clinically significant GVHD group (s-GVHD; grade 2–4 aGVHD and severe cGVHD, n=5). Markers have significant associations or trends with GVHD are exhibited. A Representative flow cytometry data from patients of each group. B Summary data of surface markers that expressed on CD4+ /CD8+ T cells shown through the box-and-whiskers plots. Each dot represents an individual patient. P values were calculated using Wilcoxon rank-sum tests and shown with raw values. Fig. S5 Gating strategies for analyzing the components of immune cells. The definition of each cell subset is listed in Table S3.