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. 2024 Jan 5;143(15):1496–1512. doi: 10.1182/blood.2023021671

Figure 1.

Figure 1.

S aureus culture supernatants and SE induce drug resistance in malignant T cells. (A-C) Photographs of affected skin from 3 patients with SS before and after antibiotic treatment with ongoing cancer-directed treatments: (A) doxorubicin (SS1), (B) vorinostat (SS2), and (C) alitretinoin (SS3). (D) Flow cytometric plots showing apoptotic fraction of malignant T cells from SS17 PBMCs after 72 hours of treatment with 2 nM romidepsin and bacterial culture supernatant (sup) from S aureus cultured in the presence or absence of endolysin. The used S aureus strain was originally isolated from lesional skin of a different patient with SS. For control, tryptic soy broth medium (Ctrl medium) was added to the PBMC culture. (E) Western blot showing cleaved and uncleaved PARP expression after 24- and 48-hour treatment of SS4 PBMCs with either SE and/or romidepsin. GAPDH is used as a loading control. (F) Apoptotic fraction of malignant cells from PBMCs of 5 patients with SS (SS4, SS5, SS8, SS9, and SS12) after 72 and 144 hours treated with increasing concentrations of romidepsin in the presence (SE) or absence (PBS) of SE. Statistical significance was assessed by 2-way analysis of variance (ANOVA) followed by the Šídák multiple comparisons test. ∗∗P < .01; ∗∗∗P < .0005. (G) Apoptotic fraction of malignant cells from PBMCs of 2 patients with SS (SS8 and SS12) after a different 6 hours pulse, 16 hours chase in vitro treatment regimen with high concentrations of romidepsin in the presence (SE) or absence (PBS) of SE. (H-I) Representative flow cytometric plots (H; SS13) and quantification (I) of apoptotic fraction of malignant cells from PBMCs of 5 patients with SS (SS4, SS8, SS12, SS13, and SS15) after 72 hours of treatment with 2 nM romidepsin in presence of wild-type SEA or mutant SEA (SEAF47A/D227A). (J-K) Representative flow cytometric plots (J; SS15) and quantification (K) of apoptotic fraction of malignant cells from 4 patients with SS (SS4, SS12, SS13, and SS15) after 72 hours of treatment with 2 nM romidepsin in presence of SEA or SEA and a blocking anti-SEA antibody. For panels I-K, statistical significance was assessed by repeated measures 1-way ANOVA followed by Tukey multiple comparison test. ∗P < .05. Ctrl, control; DMSO, dimethyl sulfoxide; GAPDH, glyceraldehyde-3-phosphate dehydrogenase.