Journal: NAR Cancer

Article Title: Co-expression of CD30 and SLFN11 serves as a dual biomarker for the treatment of cutaneous T-cell lymphoma

doi: 10.1093/narcan/zcaf037

Figure Lengend Snippet: SLFN11 expression in MF/SS and its impact on clinical outcome. ( A ) Representative images of immunohistochemical staining for CD30 in skin-infiltrating MF tumor cells (original magnification, scale bar indicating 100 μm). ( B ) Quantification of CD30-positive population (%) in MF/SS tumors using an image analysis software (StrataQuest 7). Representative CD30-negative and -positive results are shown. ( C ) Distribution of CD30-positive population (%) in all cases of MF/SS. A red line indicates a cut-off value at 5%. ( D ) Representative images of immunohistochemical staining for SLFN11 in skin-infiltrating MF tumor cells (top). Vascular endothelial cells in the same sample from Case 13 are positive for SLFN11 (bottom, arrow; original magnification, scale bar indicating 100 μm). ( E ) Quantification of SLFN11-positive population (%) in MF/SS tumors using an image analysis software (StrataQuest 7). Representative SLFN11-negative and -positive results are shown. ( F ) Distribution of SLFN11-positive population (%) in all cases of MF/SS. A red line indicates a cut-off value at 10%. ( G ) Correlation analysis between % of SLFN11-positive (SLFN11 + ) cells and % of CD30-positive (CD30 + ) cells (right) or % of SLFN11 + cells and serum values of sIL-2R (left). Each dot represents one patient. The correlation coefficient ( r ) was calculated using the Pearson correlation coefficient test. ( H ) Kaplan–Meier curves for OS rate (%) in all the enrolled MF/SS patients (25 in MF and 5 in SS patients). For stage IA–IB versus IV, hazard ratio (HR), 0.1183; 95% confidence interval (CI), 0.02969–0.4714; P = .0027 (log-rank test). For stage IIB–III versus IV, HR, 0.1041; 95% CI, 0.02491–0.4350; P = .0046. For stage IA–IB versus IIB–III, HR, 5.755; 95% CI, 0.1096–302.1; P = .3865. ( I ) Kaplan–Meier curves for OS rate (%) in 13 MF/SS patients with stage IV group (8 in SLFN11-positive group and 5 in SLFN11-negative group), HR, 0.7759; 95% CI, 0.1665–3.614; P = .3823. ( J ) Kaplan–Meier curves for OS rate (%) in 13 MF/SS patients with stage IV group (9 in CD30-positive group and 4 in CD30-negative group), HR, 0.7263; 95% CI, 0.1542–3.422; P = .8591. N.S., not significant.

Article Snippet: Sections were incubated with rabbit anti-CD30 antibody (1:200; Cell Signaling Technology, #54 535, clone E4L4I) and mouse anti-SLFN11 antibody (D-2) at RT for 1 h. After washing, the slides were incubated with matched secondary antibodies, Alexa Fluor 594 donkey anti-mouse IgG and Alexa Fluor 488 donkey anti-rabbit IgG (1:200; Thermo Fisher Scientific), for 30 min at RT, then washed and mounted with Mounting Medium with DAPI (Thermo Fisher Scientific).

Techniques: Expressing, Immunohistochemical staining, Staining, Software

Journal: NAR Cancer

Article Title: Co-expression of CD30 and SLFN11 serves as a dual biomarker for the treatment of cutaneous T-cell lymphoma

doi: 10.1093/narcan/zcaf037

Figure Lengend Snippet: Co-expression of SLFN11 and CD30 in MF tumor cells. ( A ) Representative images of immunohistochemical staining for SLFN11 and CD30 in skin-infiltrating MF tumor cells (original magnification, scale bar indicating 100 μm). ( B ) Representative images of immunofluorescence staining for SLFN11 (red), CD30 (green), and DAPI (blue) corresponding to Fig. (original magnification, scale bar indicating 100 μm). ( C ) An image cytometry analysis by StrataQuest 7 shows the intensity of SLFN11 and CD30 in each cell.

Article Snippet: Sections were incubated with rabbit anti-CD30 antibody (1:200; Cell Signaling Technology, #54 535, clone E4L4I) and mouse anti-SLFN11 antibody (D-2) at RT for 1 h. After washing, the slides were incubated with matched secondary antibodies, Alexa Fluor 594 donkey anti-mouse IgG and Alexa Fluor 488 donkey anti-rabbit IgG (1:200; Thermo Fisher Scientific), for 30 min at RT, then washed and mounted with Mounting Medium with DAPI (Thermo Fisher Scientific).

Techniques: Expressing, Immunohistochemical staining, Staining, Immunofluorescence, Cytometry

Journal: NAR Cancer

Article Title: Co-expression of CD30 and SLFN11 serves as a dual biomarker for the treatment of cutaneous T-cell lymphoma

doi: 10.1093/narcan/zcaf037

Figure Lengend Snippet: Activation via T-cell receptor/IL-2R induces the functional SLFN11 expression in normal T cells. ( A ) Immunoblotting showing SLFN11, CD30, and CD3 expressions in PBMCs with or without IFN-γ or CD3/CD28/IL-2 stimulation for 5 days. Actin was used as a loading control. ( B ) Representative confocal microscopy images for SLFN11 (green), CD3 (red), and DAPI (blue) (top panel) or SLFN11 (green), CD30 (red), and DAPI (blue) (bottom panel) in PBMCs 7 days after each activation (original magnification, scale bar indicating 10 μm). ( C ) Viability curves to various concentrations of TOP1 inhibitor CPT in PBMCs with or without CD3/CD28/IL-2 stimulation for 5–7 days. Viability was examined by ATP assay 48 h after the CPT treatments. Representative results in triplicate from two independent experiments are shown as mean ± SD.

Article Snippet: Sections were incubated with rabbit anti-CD30 antibody (1:200; Cell Signaling Technology, #54 535, clone E4L4I) and mouse anti-SLFN11 antibody (D-2) at RT for 1 h. After washing, the slides were incubated with matched secondary antibodies, Alexa Fluor 594 donkey anti-mouse IgG and Alexa Fluor 488 donkey anti-rabbit IgG (1:200; Thermo Fisher Scientific), for 30 min at RT, then washed and mounted with Mounting Medium with DAPI (Thermo Fisher Scientific).

Techniques: Activation Assay, Functional Assay, Expressing, Western Blot, Control, Confocal Microscopy, ATP Assay

Journal: NAR Cancer

Article Title: Co-expression of CD30 and SLFN11 serves as a dual biomarker for the treatment of cutaneous T-cell lymphoma

doi: 10.1093/narcan/zcaf037

Figure Lengend Snippet: JAK pathway activation and epigenetic suppression regulate SLFN11 expression in CTCL cell lines. ( A ) Immunoblotting showing SLFN11 and CD30 expressions in HUT78 cells treated with pan-JAK inhibitor, cerdulatinib (40 μM), for the indicated time. Actin was used as a loading control. A representative result of two independent experiments is shown. ( B ) Immunoblotting showing SLFN11 expression in MJ cells treated with the indicated HDAC inhibitors (10 μM each) for 16 h. GAPDH was used as a loading control. The relative ratio of SLFN11/GAPDH in each HDAC inhibitor-treated MJ cell is indicated at the bottom. A representative result of three independent experiments is shown. ( C ) Viability curves of the MJ cells to the indicated HDAC inhibitors. Viability was examined by ATP assay 72 h after the drug treatments. Representative results in triplicate from two independent experiments are shown as mean ± SD.

Article Snippet: Sections were incubated with rabbit anti-CD30 antibody (1:200; Cell Signaling Technology, #54 535, clone E4L4I) and mouse anti-SLFN11 antibody (D-2) at RT for 1 h. After washing, the slides were incubated with matched secondary antibodies, Alexa Fluor 594 donkey anti-mouse IgG and Alexa Fluor 488 donkey anti-rabbit IgG (1:200; Thermo Fisher Scientific), for 30 min at RT, then washed and mounted with Mounting Medium with DAPI (Thermo Fisher Scientific).

Techniques: Activation Assay, Expressing, Western Blot, Control, ATP Assay