Journal: Redox Biology

Article Title: Systolic pressure overload caused pulmonary oxidative stress, vessel remodeling and severe microvascular thrombosis in CD40 knockout mice through promoting platelet aggregation

doi: 10.1016/j.redox.2026.104122

Figure Lengend Snippet: TAC caused similar left ventricular hypertrophy, fibrosis and accumulation of immune cells in WT and CD40 KO mice. Post-TAC survival analysis of WT (n = 41) and CD40 KO (n = 37) mice (A). The ratios of LV and LA weight to tibial length of WT and CD40 KO mice under control or TAC condition (n = 9-13), Sham indicates no actual TAC (B, C). Representative images and summary data for LV myocyte cross-sectional area determined by FITC-conjugated wheat germ agglutinin (WGA) staining (n = 5-7) (D). Representative images and quantitative data of LV fibrosis by Sirius red/Fast green staining (n = 4-7) (E). CD45 immunostaining (red) and quantitative data of LV leukocyte infiltration (n = 5-6) (F). Survival rate was analyzed by Kaplan-Meier method and compared by log-rank test. All quantitative data are reported as means ± SEM. Data were analyzed using one-way ANOVA followed by Bonferroni post hoc analysis. ns indicates nonsignificant ( p > 0.05),∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001,∗∗∗∗ p < 0.0001.

Article Snippet: Animals and experimental design: CD40 KO mice (B6.129P2– Cd40 tm1Kik /J; stock NO: 002928) and wild type (WT) C57BL/6J mice were purchased from Jackson Laboratory and Shanghai SLAC Laboratory Animal Co, Ltd. Based on the information provided by Jackson Laboratory, CD40 KO strain has been backcrossed to C57BL/6J mice for at least 10 generations.

Techniques: Control, Staining, Immunostaining

Journal: Redox Biology

Article Title: Systolic pressure overload caused pulmonary oxidative stress, vessel remodeling and severe microvascular thrombosis in CD40 knockout mice through promoting platelet aggregation

doi: 10.1016/j.redox.2026.104122

Figure Lengend Snippet: TAC caused comparable LV dysfunction in WT and CD40 KO mice. Representative echocardiograms of each group (A). Summary data for LV ejection fraction (EF%), fractional shortening (FS%), LV end-diastolic and end-systolic diameter (LV-EDD, LV-ESD) and heart rate (n = 9-13) in each group (B–F). LV pressure of WT (n = 5) and CD40 KO (n = 5) mice were measured at 8 weeks after TAC, representative invasive pressure curve(G) with quantifications of end-systolic pressure (ESP)(I) and end-diastolic pressure (EDP)(J). Representative dp/dt curve(H) with quantifications of dp/dtmax (maximal rate of change in systolic pressure over time) (L) and dp/dtmin (minimal rate of change in pressure over time) (M). The rate-pressure product (K) was calculated by multiplying the heart rate by the LV end-systolic pressure. Western blot of β-MHC and vinculin (loading control, n = 3-4) (N, O). All quantitative data are reported as mean ± SEM. The statistical significance was assessed using two-tailed Student's unpaired t tests(I-M). Data were analyzed using one-way ANOVA followed by Bonferroni post hoc analysis (B–F, N, O). ns indicates nonsignificant ( p > 0.05),∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001,∗∗∗∗ p < 0.0001.

Article Snippet: Animals and experimental design: CD40 KO mice (B6.129P2– Cd40 tm1Kik /J; stock NO: 002928) and wild type (WT) C57BL/6J mice were purchased from Jackson Laboratory and Shanghai SLAC Laboratory Animal Co, Ltd. Based on the information provided by Jackson Laboratory, CD40 KO strain has been backcrossed to C57BL/6J mice for at least 10 generations.

Techniques: Western Blot, Control, Two Tailed Test

Journal: Redox Biology

Article Title: Systolic pressure overload caused pulmonary oxidative stress, vessel remodeling and severe microvascular thrombosis in CD40 knockout mice through promoting platelet aggregation

doi: 10.1016/j.redox.2026.104122

Figure Lengend Snippet: CD40 KO aggravated TAC-induced increase of lung weight. The ratios of lung wet weight, lung dry weight, lung water weight to tibial length (A-C). Lung water content percentage ((lung wet weight – lung dry weight)/lung wet weight × 100%) of each group mice (n = 7-13) (D). All quantitative data are reported as mean ± SEM. Data were analyzed using one-way ANOVA followed by Bonferroni post hoc analysis. ns indicates nonsignificant ( p > 0.05),∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001,∗∗∗∗ p < 0.0001.

Article Snippet: Animals and experimental design: CD40 KO mice (B6.129P2– Cd40 tm1Kik /J; stock NO: 002928) and wild type (WT) C57BL/6J mice were purchased from Jackson Laboratory and Shanghai SLAC Laboratory Animal Co, Ltd. Based on the information provided by Jackson Laboratory, CD40 KO strain has been backcrossed to C57BL/6J mice for at least 10 generations.

Techniques:

Journal: Redox Biology

Article Title: Systolic pressure overload caused pulmonary oxidative stress, vessel remodeling and severe microvascular thrombosis in CD40 knockout mice through promoting platelet aggregation

doi: 10.1016/j.redox.2026.104122

Figure Lengend Snippet: CD40 KO caused robust pulmonary microvascular thrombosis in mice after TAC. Representative images of hematoxylin and eosin (HE) staining of lung tissues from the experimental groups(A) . Representative immunostaining images of Carstairs staining of the lung (Fibrin: bright red, platelets: gray blue or navy, collagen: bright blue), CD42c, CD61 and vWF in the lung (B-E). Quantitative data of CD42c immunofluorescence staining in the lung (n = 5-7) (F). The platelet counts in each group of mice (n = 8-14) (G). Mean platelet volume (n = 8-14) (H). Quantitative data of vWF in the lung (n = 5-6) (I). The CD42c and vWF staining were expressed as a percentage of positive staining area to the total area. All quantitative data are reported as mean ± SEM. Data were analyzed using one-way ANOVA followed by Bonferroni post hoc analysis. ns indicates nonsignificant ( p > 0.05),∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001,∗∗∗∗ p < 0.0001.

Article Snippet: Animals and experimental design: CD40 KO mice (B6.129P2– Cd40 tm1Kik /J; stock NO: 002928) and wild type (WT) C57BL/6J mice were purchased from Jackson Laboratory and Shanghai SLAC Laboratory Animal Co, Ltd. Based on the information provided by Jackson Laboratory, CD40 KO strain has been backcrossed to C57BL/6J mice for at least 10 generations.

Techniques: Staining, Immunostaining, Immunofluorescence

Journal: Redox Biology

Article Title: Systolic pressure overload caused pulmonary oxidative stress, vessel remodeling and severe microvascular thrombosis in CD40 knockout mice through promoting platelet aggregation

doi: 10.1016/j.redox.2026.104122

Figure Lengend Snippet: CD40 KO exacerbated platelet aggregation and blood clot retraction in mice after TAC. The aggregation levels of WT and CD40 KO mouse platelets in response to 1.2 μg/ml collagen stimulation (n = 3-5) (A). The aggregation levels of WT and CD40 KO mouse platelets in response to 0.067U/ml thrombin stimulation (n = 5-6) (B). The aggregation levels of WT and CD40 KO mouse platelets in response to 3.4U/ml ADP stimulation (n = 4-5) (C). Clot retraction of WT and CD40 KO mouse platelets (n = 5-7) (D). All quantitative data are reported as mean ± SEM. Data were analyzed using one-way ANOVA followed by Bonferroni post hoc analysis. ns indicates nonsignificant ( p > 0.05),∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001,∗∗∗∗ p < 0.0001.

Article Snippet: Animals and experimental design: CD40 KO mice (B6.129P2– Cd40 tm1Kik /J; stock NO: 002928) and wild type (WT) C57BL/6J mice were purchased from Jackson Laboratory and Shanghai SLAC Laboratory Animal Co, Ltd. Based on the information provided by Jackson Laboratory, CD40 KO strain has been backcrossed to C57BL/6J mice for at least 10 generations.

Techniques:

Journal: Redox Biology

Article Title: Systolic pressure overload caused pulmonary oxidative stress, vessel remodeling and severe microvascular thrombosis in CD40 knockout mice through promoting platelet aggregation

doi: 10.1016/j.redox.2026.104122

Figure Lengend Snippet: CD40 KO aggravated TAC-induced pulmonary leukocyte infiltration. Representative images and quantitative data of CD45 + cells, Mac2 + cells and CD3 + cells in lungs (n = 5-7) (A-C). All quantitative data are reported as mean ± SEM. Data were analyzed using one-way ANOVA followed by Bonferroni post hoc analysis. ns indicates nonsignificant ( p > 0.05),∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001,∗∗∗∗ p < 0.0001.

Article Snippet: Animals and experimental design: CD40 KO mice (B6.129P2– Cd40 tm1Kik /J; stock NO: 002928) and wild type (WT) C57BL/6J mice were purchased from Jackson Laboratory and Shanghai SLAC Laboratory Animal Co, Ltd. Based on the information provided by Jackson Laboratory, CD40 KO strain has been backcrossed to C57BL/6J mice for at least 10 generations.

Techniques:

Journal: Redox Biology

Article Title: Systolic pressure overload caused pulmonary oxidative stress, vessel remodeling and severe microvascular thrombosis in CD40 knockout mice through promoting platelet aggregation

doi: 10.1016/j.redox.2026.104122

Figure Lengend Snippet: CD40 KO exacerbated pulmonary fibrosis and vessel remodeling in mice after TAC. Representative images (A-C) and quantitative data (D-F) of Masson's trichrome staining and smooth muscle α-actin (red) of the lung tissues. Quantitative RT-PCR result of pulmonary TGFβ (normalized to 18S) (G). All quantitative data are reported as mean ± SEM. Data were analyzed using one-way ANOVA followed by Bonferroni post hoc analysis. ns indicates nonsignificant ( p > 0.05),∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001,∗∗∗∗ p < 0.0001.

Article Snippet: Animals and experimental design: CD40 KO mice (B6.129P2– Cd40 tm1Kik /J; stock NO: 002928) and wild type (WT) C57BL/6J mice were purchased from Jackson Laboratory and Shanghai SLAC Laboratory Animal Co, Ltd. Based on the information provided by Jackson Laboratory, CD40 KO strain has been backcrossed to C57BL/6J mice for at least 10 generations.

Techniques: Staining, Quantitative RT-PCR

Journal: Redox Biology

Article Title: Systolic pressure overload caused pulmonary oxidative stress, vessel remodeling and severe microvascular thrombosis in CD40 knockout mice through promoting platelet aggregation

doi: 10.1016/j.redox.2026.104122

Figure Lengend Snippet: CD40 KO exacerbated pulmonary oxidative stress in mice with existing LV dysfunction. Representative images and Quantitative data of pulmonary dihydroethidium (DHE) staining in mice (n = 5) (A, D), IHC analysis of 3′-nitrotyrosine (B, E) and 4-hydroxynonenal (n = 4-6) (C, F). All quantitative data are reported as mean ± SEM. Data were analyzed using one-way ANOVA followed by Bonferroni post hoc analysis. ns indicates nonsignificant ( p > 0.05),∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001,∗∗∗∗ p < 0.0001.

Article Snippet: Animals and experimental design: CD40 KO mice (B6.129P2– Cd40 tm1Kik /J; stock NO: 002928) and wild type (WT) C57BL/6J mice were purchased from Jackson Laboratory and Shanghai SLAC Laboratory Animal Co, Ltd. Based on the information provided by Jackson Laboratory, CD40 KO strain has been backcrossed to C57BL/6J mice for at least 10 generations.

Techniques: Staining

Journal: Redox Biology

Article Title: Systolic pressure overload caused pulmonary oxidative stress, vessel remodeling and severe microvascular thrombosis in CD40 knockout mice through promoting platelet aggregation

doi: 10.1016/j.redox.2026.104122

Figure Lengend Snippet: TAC resulted in significant RV inflammation, fibrosis and cardiomyocyte hypertrophy in CD40 KO mice. The ratio of RV weight to tibial length of WT and CD40 KO mice under control or TAC condition (n = 9-13) (A). Western blot of β-MHC and loading control of vinculin (n = 3-4) (B, C)). Representative images and summary data for RV myocyte cross-sectional area determined by FITC-conjugated WGA staining (n = 6) (D). Representative images and quantitative data of RV fibrosis by Sirius red/Fast green staining (n = 5-6) (E). CD45 immunostaining (red) and quantitative data of RV leukocyte infiltration (n = 5-7) (F). All quantitative data are reported as mean ± SEM. Data were analyzed using one-way ANOVA followed by Bonferroni post hoc analysis. ns indicates nonsignificant ( p > 0.05),∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001,∗∗∗∗ p < 0.0001.

Article Snippet: Animals and experimental design: CD40 KO mice (B6.129P2– Cd40 tm1Kik /J; stock NO: 002928) and wild type (WT) C57BL/6J mice were purchased from Jackson Laboratory and Shanghai SLAC Laboratory Animal Co, Ltd. Based on the information provided by Jackson Laboratory, CD40 KO strain has been backcrossed to C57BL/6J mice for at least 10 generations.

Techniques: Control, Western Blot, Staining, Immunostaining

Journal: Science Advances

Article Title: Inflammatory arthritis irAE may represent a unique autoimmune disease primarily driven by T cells but likely not autoantibodies

doi: 10.1126/sciadv.aea4262

Figure Lengend Snippet: ( A to C ) Human naïve B cells were isolated from PBMCs and cultured under indicated conditions for 7 days; pembrolizumab or isotype control IgG4 (IgG4 Iso) was added on day 2; n = 8. (A) Expression of CD38 and CD27 on B cells. Right: Percentages of CD27 + CD38 − and CD27 + CD38 + B cells. (B) Expression of CD138 on CD27 + CD38 + B cells. Right: Percentage of CD138 + CD27 + CD38 + B cells. (C) Different immunoglobulin isotypes were measured in the culture supernatant of (A) and (B) by multiplex assay. ( D to F ) B cells from Humanized PD-1 (HuPD-1) mice were isolated and cultured with LPS, IL-4, BAFF, or ODN 2006, anti-IgM, IL-21, IL-4 or R848, anti-IgM, anti-CD40, IL-21, and IFN-γ for 3 days; pembrolizumab or isotype control was added on day 1. (D) Expression of IgG2c on activated B cells. FSC-H, Forward scatter height. Right: Percentage of IgG2c + B cells; n = 5. (E) Expression of IgG1 on activated B cells. Right: Percentage of IgG1 + B cells; n = 5. (F) Different immunoglobulin isotypes in the supernatant were measured by multiplex assay; n = 5. Data in graphs represent mean ± SEM. Significance was tested by two-way ANOVA.

Article Snippet: Condition 3 ( ): R848 (1 μg/ml), anti-mouse CD40 (1 μg/ml; Bio X Cell, catalog no. BE0016-2), anti-mouse IgM (1 μg/ml), rmIL-21 (100 ng/ml), rmIFN-γ (10 ng/ml; Peprotech, catalog no. 315-05; 20 μg).

Techniques: Isolation, Cell Culture, Control, Expressing, Multiplex Assay

Journal: Science Advances

Article Title: Inflammatory arthritis irAE may represent a unique autoimmune disease primarily driven by T cells but likely not autoantibodies

doi: 10.1126/sciadv.aea4262

Figure Lengend Snippet: ( A to F ) Bead-based multiplex assays were used to measure plasma concentration of IL-6, and IL-12p70 (A); TNF-α, IFN-γ, and IL-1β [B; HC ( n = 19), irAE ( n = 34), RAC ( n = 45), and ICI ( n = 9)]; IP-10 (CXCL10), CXCL11, and CXCL9 (C; HC, n = 17; irAE, n = 33; RAC, n = 46; ICI, n = 17); CCL20 (D); CX3CL1 (E); and CCL2 (F). ( G to J ) Human naïve B cells were isolated and cultured with anti–human CD40 (0.5 μg/ml), anti–human Ig (M + G + A) (2.5 μg/ml), and rhIL-21 (20 ng/ml) with IFN-α (100 ng/ml), IL-6 (100 ng/ml), IL-12 (100 ng/ml), control, or the combination of IFN-α, IL-6, and IL-12 for 7 days. Cells and culture supernatants were analyzed. (G) Representative flow plot of CD38 and CD138 expression on CD27 hi CD38 hi ASCs. Right: A summary of the percentage of CD138 + ASCs; n = 6. (H) Expression of CD11c and CD27 on CD27 − IgD − B cells. Right: Percentage of CD11c + IgD − CD27 − B cells; n = 6. (I) Expression of active-caspase-3 in B cells. Right: Percentage of active-caspase-3 + B cells from different groups; n = 3. (J) Different immunoglobulin isotype levels in the culture supernatants from (G) to (H) were measured by the multiplex assay; n = 6. Data in graphs represent mean ± SEM. Significance was tested by one-way ANOVA [(A) to (I)] and paired Student’s t test (J). [(A) to (F)] ICI, ICI control.

Article Snippet: Condition 3 ( ): R848 (1 μg/ml), anti-mouse CD40 (1 μg/ml; Bio X Cell, catalog no. BE0016-2), anti-mouse IgM (1 μg/ml), rmIL-21 (100 ng/ml), rmIFN-γ (10 ng/ml; Peprotech, catalog no. 315-05; 20 μg).

Techniques: Multiplex Assay, Clinical Proteomics, Concentration Assay, Isolation, Cell Culture, Control, Expressing

Journal: Science Advances

Article Title: Inflammatory arthritis irAE may represent a unique autoimmune disease primarily driven by T cells but likely not autoantibodies

doi: 10.1126/sciadv.aea4262

Figure Lengend Snippet: ( A to C ) Human naïve B cells were isolated from PBMCs and cultured under indicated conditions for 7 days; pembrolizumab or isotype control IgG4 (IgG4 Iso) was added on day 2; n = 8. (A) Expression of CD38 and CD27 on B cells. Right: Percentages of CD27 + CD38 − and CD27 + CD38 + B cells. (B) Expression of CD138 on CD27 + CD38 + B cells. Right: Percentage of CD138 + CD27 + CD38 + B cells. (C) Different immunoglobulin isotypes were measured in the culture supernatant of (A) and (B) by multiplex assay. ( D to F ) B cells from Humanized PD-1 (HuPD-1) mice were isolated and cultured with LPS, IL-4, BAFF, or ODN 2006, anti-IgM, IL-21, IL-4 or R848, anti-IgM, anti-CD40, IL-21, and IFN-γ for 3 days; pembrolizumab or isotype control was added on day 1. (D) Expression of IgG2c on activated B cells. FSC-H, Forward scatter height. Right: Percentage of IgG2c + B cells; n = 5. (E) Expression of IgG1 on activated B cells. Right: Percentage of IgG1 + B cells; n = 5. (F) Different immunoglobulin isotypes in the supernatant were measured by multiplex assay; n = 5. Data in graphs represent mean ± SEM. Significance was tested by two-way ANOVA.

Article Snippet: Condition 1: anti–human Ig (M + G + A) (2.5 μg/ml; Jackson ImmunoResearch, catalog no. 109-006-064), CpG oligodeoxynucleotide (ODN) (2.5 μg/ml; Invivogen, catalog no. tlrl-2006-1), anti–human CD40 (10 μg/ml; Bio X Cell, catalog no. BE0189), rhIL-21 (20 ng/ml; Peprotech, catalog no. 200-21-50UG), rhIL-4 (10 ng/ml; BioLegend, catalog no. 574004), and rhIL-2 (10 ng/ml; Peprotech, catalog no. 200-02-250UG).

Techniques: Isolation, Cell Culture, Control, Expressing, Multiplex Assay

Journal: Science Advances

Article Title: Inflammatory arthritis irAE may represent a unique autoimmune disease primarily driven by T cells but likely not autoantibodies

doi: 10.1126/sciadv.aea4262

Figure Lengend Snippet: ( A to F ) Bead-based multiplex assays were used to measure plasma concentration of IL-6, and IL-12p70 (A); TNF-α, IFN-γ, and IL-1β [B; HC ( n = 19), irAE ( n = 34), RAC ( n = 45), and ICI ( n = 9)]; IP-10 (CXCL10), CXCL11, and CXCL9 (C; HC, n = 17; irAE, n = 33; RAC, n = 46; ICI, n = 17); CCL20 (D); CX3CL1 (E); and CCL2 (F). ( G to J ) Human naïve B cells were isolated and cultured with anti–human CD40 (0.5 μg/ml), anti–human Ig (M + G + A) (2.5 μg/ml), and rhIL-21 (20 ng/ml) with IFN-α (100 ng/ml), IL-6 (100 ng/ml), IL-12 (100 ng/ml), control, or the combination of IFN-α, IL-6, and IL-12 for 7 days. Cells and culture supernatants were analyzed. (G) Representative flow plot of CD38 and CD138 expression on CD27 hi CD38 hi ASCs. Right: A summary of the percentage of CD138 + ASCs; n = 6. (H) Expression of CD11c and CD27 on CD27 − IgD − B cells. Right: Percentage of CD11c + IgD − CD27 − B cells; n = 6. (I) Expression of active-caspase-3 in B cells. Right: Percentage of active-caspase-3 + B cells from different groups; n = 3. (J) Different immunoglobulin isotype levels in the culture supernatants from (G) to (H) were measured by the multiplex assay; n = 6. Data in graphs represent mean ± SEM. Significance was tested by one-way ANOVA [(A) to (I)] and paired Student’s t test (J). [(A) to (F)] ICI, ICI control.

Article Snippet: Condition 1: anti–human Ig (M + G + A) (2.5 μg/ml; Jackson ImmunoResearch, catalog no. 109-006-064), CpG oligodeoxynucleotide (ODN) (2.5 μg/ml; Invivogen, catalog no. tlrl-2006-1), anti–human CD40 (10 μg/ml; Bio X Cell, catalog no. BE0189), rhIL-21 (20 ng/ml; Peprotech, catalog no. 200-21-50UG), rhIL-4 (10 ng/ml; BioLegend, catalog no. 574004), and rhIL-2 (10 ng/ml; Peprotech, catalog no. 200-02-250UG).

Techniques: Multiplex Assay, Clinical Proteomics, Concentration Assay, Isolation, Cell Culture, Control, Expressing