Journal: Diabetologia

Article Title: Phenotypically distinct anti-insulin B cells repopulate pancreatic islets after anti-CD20 treatment in NOD mice

doi: 10.1007/s00125-019-04974-y

Figure Lengend Snippet: Anti-insulin B cells repopulate pancreatic islets more rapidly than insulin-negative B cells after anti-CD20 treatment. Groups of VH125.hCD20/NOD mice, aged 6–8 weeks old, were injected with 2H7 anti-CD20 or IgG isotype control. Spleen, PLNs and pancreatic islets were analysed for insulin-positive and insulin-negative B cells at 8 weeks and 12 weeks post depletion by flow cytometry. ( a – f ) No. of cells from IgG control-treated (black circles) and 2H7-treated (grey squares) mice for insulin-negative B cells ( a – c ) and insulin-positive B cells ( d – f ) from spleen ( a , d ) PLNs ( b , e ) and islets ( c , f ). ( g – i ) Percentage of B cells repopulated at 8 and 12 weeks after treatment from spleen ( g ), PLNs ( h ) and islets ( i ) of mice shown in ( a – f ). Percentages were calculated as individual numbers from each 2H7-treated mouse / mean number from all IgG control antibody-treated mice. Horizontal lines represent the median value. Data represent three independent experiments. At 8 weeks, n = 7 (spleen), n = 7 (PLNs) and n = 9 (islets) for control IgG-treated mice and n = 10 (spleen), n = 9 (PLNs) and n = 12 (islets) for 2H7-treated mice. At 12 weeks, n = 8 (spleen), n = 6 (PLNs) and n = 7 (islets) for control IgG and n = 11 (spleen), n = 7 (PLNs) and n = 11 (islets) for 2H7-treated mice. * p < 0.05 (one-way ANOVA)

Article Snippet: Female VH125.hCD20/NOD mice, 6–8 weeks of age were chosen at random to receive either anti-hCD20 antibody (clone 2H7; Bio-XCell, West Lebanon, NH, USA) or control IgG2b antibody (clone MPC-11; Bio-XCell [ , , ]), as described previously [ ].

Techniques: Injection, Control, Flow Cytometry

Journal: Diabetologia

Article Title: Phenotypically distinct anti-insulin B cells repopulate pancreatic islets after anti-CD20 treatment in NOD mice

doi: 10.1007/s00125-019-04974-y

Figure Lengend Snippet: CD138 int anti-insulin B cells are enriched in pancreatic islets after anti-CD20 treatment. Groups of 6- to 8-week-old VH125.hCD20/NOD mice were injected with 2H7 anti-CD20 or IgG isotype control. Groups of mice ( n = 2 or 3 per group) were pooled and insulin + B cells from pancreatic islets were analysed for four different populations based on CD138 expression: CD138 − (blue); CD138 int IgM + (orange); CD138 int IgM lo (grey) and CD138 hi IgM lo (red). ( a , b ) Representative flow plots showing gating on live CD3 − CD11b − CD11c − ( a ) and graph showing the overall percentages of the four different populations ( b ). ( c , d ) Representative flow plots showing insulin − CD19 + , insulin + CD19 + and insulin + CD19 − cells ( c ) and graph showing the overall percentages of these cells ( d ); 2H7 (black circles), IgG (grey circles). ( e ) Representative flow plots showing CD138 and IgM expression in insulin + CD19 + and insulin + CD19 − cells. ( f , g ) Graphs showing CD138 and IgM populations on insulin + CD19 + ( f ) and insulin + CD19 − cells ( g ) ( n = 5 groups for control IgG treatment; n = 4 groups for 2H7 treatment). Horizontal lines represent the median values. Data represent two independent experiments. * p < 0.05 (one-way ANOVA)

Article Snippet: Female VH125.hCD20/NOD mice, 6–8 weeks of age were chosen at random to receive either anti-hCD20 antibody (clone 2H7; Bio-XCell, West Lebanon, NH, USA) or control IgG2b antibody (clone MPC-11; Bio-XCell [ , , ]), as described previously [ ].

Techniques: Injection, Control, Expressing

Journal: Discover oncology

Article Title: VSV-CHIKV activates antitumor immunity by inducing pyroptosis in a melanoma model.

doi: 10.1007/s12672-025-02788-6

Figure Lengend Snippet: Fig. 1 Oncolytic VSV-CHIKV induces melanoma cell death (MOI = 1, 24 h). A Schematic showing the wild-type VSV and chimeric VSV-CHIKV genomes, in which the VSV glycoprotein G gene (blue) is replaced by the chikungunya glycoprotein sequence in the CHIKV structural poly- protein (green). B Images obtained by microscopy showing changes in VSV-CHIKV infection of B16 cells (scale bar: 20 μm). C LDH release from the culture supernatant of B16 cells infected with VSV-CHIKV (n = 3). ***P < 0.001 by t test. D Fluorescence microscopy observation of PI uptake in B16 cells (scale bars: 50 μm). ***P < 0.001 by t test. E The levels of IL-1β and IL-18 in the cell culture supernatant (n = 3). ***P < 0.001 by t test. F Western blot analysis of NLRP3, Caspase1, and Cleaved-caspase1 protein expression after the addition of VSV-CHIKV

Article Snippet: In brief, after deparaffinization, hydration and blockage of endogenous peroxidase, the sections were incubated with 5% goat serum to block specific sites and then incubated overnight with the following primary antibodies: anti-NLRP3 antibody (68102-1-Ig, Proteintech, USA), anti-Caspase1 antibody (sc-56036, Santa Cruz Biotechnology, USA), anti-cleaved caspase 1 antibody (AF4022, Affinity, USA), anti-GSDMD antibody (ab219800, Abcam, UK) and antiGSDMD-N antibody (DF13758, Affinity, USA).

Techniques: Sequencing, Microscopy, Infection, Fluorescence, Cell Culture, Western Blot, Expressing

Journal: Discover oncology

Article Title: VSV-CHIKV activates antitumor immunity by inducing pyroptosis in a melanoma model.

doi: 10.1007/s12672-025-02788-6

Figure Lengend Snippet: Fig. 3 Oncolytic VSV-CHIKV causes pyroptosis in a melanoma cancer model. A Schematic illustration of the experimental design. B16 cell tumor-bearing C57BL/6 mice were intratumorally treated with VSV-CHIKV. B B16 tumors collected after VSV-CHIKV infection; images of the tumors, tumors weights (n = 6), body weight (n = 6), tumors growth curves are shown. **P < 0.01 by t test. ***P < 0.001 by t test. C IL-1β and IL-18 levels in tumor tissues (n = 3). ***P < 0.001 by t test. D NLRP3, Caspase1, Cleaved-caspase1, GSDMD and GSDMD-N proteins in the mock group and the VSV-CHIKV group determined by Western blotting. E Immunohistochemical staining for the NLRP3, Caspase1, Cleaved-cas- pase1, GSDMD and GSDMD-N proteins in the mock group and the VSV-CHIKV group (scale bar: 100 μm). F HE staining of major organs (scale bar: 100 μm)

Article Snippet: In brief, after deparaffinization, hydration and blockage of endogenous peroxidase, the sections were incubated with 5% goat serum to block specific sites and then incubated overnight with the following primary antibodies: anti-NLRP3 antibody (68102-1-Ig, Proteintech, USA), anti-Caspase1 antibody (sc-56036, Santa Cruz Biotechnology, USA), anti-cleaved caspase 1 antibody (AF4022, Affinity, USA), anti-GSDMD antibody (ab219800, Abcam, UK) and antiGSDMD-N antibody (DF13758, Affinity, USA).

Techniques: Infection, Western Blot, Immunohistochemical staining, Staining

Journal: Discover oncology

Article Title: VSV-CHIKV activates antitumor immunity by inducing pyroptosis in a melanoma model.

doi: 10.1007/s12672-025-02788-6

Figure Lengend Snippet: Fig. 4 Inhibition of GSDMD impairs VSV-CHIKV-induced melanoma tumor suppression. A Schematic illustration of the experimental design. B16 cell tumor-bearing C57BL/6 mice were intratumorally treated with VSV-CHIKV, VSV-CHIKV + Vehicle, or VSV-CHIKV + LDC7559. B B16 tumors from the VSV-CHIKV, VSV-CHIKV + Vehicle, and VSV-CHIKV + LDC7559 groups were collected after treatment; images of the tumors, tumors weights (n = 6), body weight (n = 6) and tumors growth curves are shown. ***P < 0.001 by One-way ANOVA. C The levels of IL-1β and IL-18 in tumor tissues (n = 3). ***P < 0.001 by One-way ANOVA. D Expression of the NLRP3, Caspase1, Cleaved-caspase1, GSDMD and GSDMD-N proteins in the VSV-CHIKV, VSV-CHIKV + Vehicle and VSV-CHIKV + LDC7559 groups determined by Western blotting. E Immunohis- tochemical staining for the NLRP3, Caspase1, Cleaved-caspase1, GSDMD and GSDMD-N proteins in the VSV-CHIKV, VSV-CHIKV + Vehicle and VSV-CHIKV + LDC7559 groups (scale bar: 100 μm). F HE staining of major organs (scale bar: 100 μm)

Article Snippet: In brief, after deparaffinization, hydration and blockage of endogenous peroxidase, the sections were incubated with 5% goat serum to block specific sites and then incubated overnight with the following primary antibodies: anti-NLRP3 antibody (68102-1-Ig, Proteintech, USA), anti-Caspase1 antibody (sc-56036, Santa Cruz Biotechnology, USA), anti-cleaved caspase 1 antibody (AF4022, Affinity, USA), anti-GSDMD antibody (ab219800, Abcam, UK) and antiGSDMD-N antibody (DF13758, Affinity, USA).

Techniques: Inhibition, Expressing, Western Blot, Staining

Journal: Discover oncology

Article Title: VSV-CHIKV activates antitumor immunity by inducing pyroptosis in a melanoma model.

doi: 10.1007/s12672-025-02788-6

Figure Lengend Snippet: Fig. 7 VSV-CHIKV induces GSDMD-mediated pyroptosis and activates antitumor immunity. The NLRP3/Caspase-1/GSDMD axis was acti- vated after melanoma cell lines were infected with VSV-CHIKV. VSV-CHIKV-triggered GSDMD-mediated tumor pyroptosis recruits CTLs into the tumor microenvironment, which is accompanied by the release of inflammatory mediators. This remodels the tumor microenvironment and turns immunologically “cold” tumors into “hot” tumors, thereby sensitizes these tumors to checkpoint blockade

Article Snippet: In brief, after deparaffinization, hydration and blockage of endogenous peroxidase, the sections were incubated with 5% goat serum to block specific sites and then incubated overnight with the following primary antibodies: anti-NLRP3 antibody (68102-1-Ig, Proteintech, USA), anti-Caspase1 antibody (sc-56036, Santa Cruz Biotechnology, USA), anti-cleaved caspase 1 antibody (AF4022, Affinity, USA), anti-GSDMD antibody (ab219800, Abcam, UK) and antiGSDMD-N antibody (DF13758, Affinity, USA).

Techniques: Infection