Journal: Cell Stem Cell

Article Title: Cyclosporine H Overcomes Innate Immune Restrictions to Improve Lentiviral Transduction and Gene Editing In Human Hematopoietic Stem Cells

doi: 10.1016/j.stem.2018.10.008

Figure Lengend Snippet:

Article Snippet: Anti-human CD38 PeVio770 , Miltenyi Biotec , Cat# 130-099-151; RRID: AB_2660384.

Techniques: Blocking Assay, Purification, Control, Recombinant, Cell Isolation, Isolation, shRNA, Software, Plasmid Preparation, Real-time Polymerase Chain Reaction

Journal: Scientific Reports

Article Title: CD38 modulates cytokine secretion by NK cells through the Sirt1/NF-κB pathway, suppressing immune surveillance in colorectal cancer

doi: 10.1038/s41598-024-79008-8

Figure Lengend Snippet: The proportions of CD38 + NK cells and Tregs in the peripheral blood of cancer patients. ( A ) Gating strategies for CD38 + NK and CD38 + NK-like T cells. CD38 + NK cells were gated as CD3- CD56 + CD38+, and CD38 + NK-like T cells were gated as CD3 + CD56 + CD38+. Proportions of CD38 + NK cells out of all NK cells in the peripheral blood of HCs ( n = 144) and patients with LC ( n = 93) ( B ), EC ( n = 37) ( C ) and CRC ( n = 126) were measured ( D ). E. Gating strategy for Tregs. The Tregs were gated as CD4 + CD25 + FOXP3+. ( F) Proportions of Tregs in the peripheral blood of HCs and CRC patients. ( G) Correlation between CD38 + NK cells and Tregs in CRC. HCs: healthy controls, LC: lung cancer, EC: esophageal cancer, CRC: colorectal cancer. *: P < 0.05, **: P < 0.01, ***: P < 0.001, and ****: P < 0.0001.

Article Snippet: CD38 + NK cells were incubated with a CD38 mAb (Proteintech) at final concentrations of 0, 1, 5 and 10 μg/ml at 37 °C in 5% CO 2 .

Techniques:

Journal: Scientific Reports

Article Title: CD38 modulates cytokine secretion by NK cells through the Sirt1/NF-κB pathway, suppressing immune surveillance in colorectal cancer

doi: 10.1038/s41598-024-79008-8

Figure Lengend Snippet: Representative immunofluorescent images of CD38 + NK cells and Tregs in tumor tissues and normal colorectal tissues. Immunofluorescence of CD38 + NK cells (CD3- CD56 + CD38+) ( A ) and Tregs (CD4 + FOXP3+) ( B ) in tumor tissues and adjacent normal tissues from CRC patients. Magnification: 10x and 40x. The proportions of CD38 + NK cells ( C ) or Tregs ( D ) in tumor tissues and adjacent normal tissues were compared. Five fields were collected from each of the five samples. **: P < 0.01 and ***: P < 0.001.

Article Snippet: CD38 + NK cells were incubated with a CD38 mAb (Proteintech) at final concentrations of 0, 1, 5 and 10 μg/ml at 37 °C in 5% CO 2 .

Techniques: Immunofluorescence

Journal: Scientific Reports

Article Title: CD38 modulates cytokine secretion by NK cells through the Sirt1/NF-κB pathway, suppressing immune surveillance in colorectal cancer

doi: 10.1038/s41598-024-79008-8

Figure Lengend Snippet: The effect of CD38 + NK cells on CD4 + T-cell differentiation. ( A ) CD38 + NK cells from HCs or CRC patients were pretreated with C-IgG or a CD38 mAb and cocultured with CD4 + T cells in transwell chambers. Representative images showing the effects of CD38 + NK cells from different groups on the differentiation of CD4 + T cells into Tregs. ( B ) Proportions of Tregs and Th1, Th2, and Th17 cells and the Th17/Treg and Th1/Th2 ratios in the CD4 + T-cell population after coculture. ( C ) Cytokine levels in the culture supernatants of CD38 + NK cells subjected to different treatments. CD38 + NK (HC): CD38 + NK cells isolated from the blood of healthy controls; CD38 + NK (CRC): CD38 + NK cells isolated from the blood of CRC patients; CRC: colorectal cancer. ns: not statistically significant; *: P < 0.05, **: P < 0.01 and ***: P < 0.001.

Article Snippet: CD38 + NK cells were incubated with a CD38 mAb (Proteintech) at final concentrations of 0, 1, 5 and 10 μg/ml at 37 °C in 5% CO 2 .

Techniques: Cell Differentiation, Isolation

Journal: Scientific Reports

Article Title: CD38 modulates cytokine secretion by NK cells through the Sirt1/NF-κB pathway, suppressing immune surveillance in colorectal cancer

doi: 10.1038/s41598-024-79008-8

Figure Lengend Snippet: The effect of CD38 on the expression of Sirt1, NF-κB and acetyl-NF-κB in NK cells. (A) Gene expression levels of Sirt1, CD38, and NF-κB in CD38 + NK cells were determined via real-time PCR. ( B ) Protein expression levels of CD38, Sirt1, NF-κB, and acetyl-NF-κB were measured via Western blot analysis. Quantitation of the protein expression was performed for CD38 ( C ) Sirt1 ( D ), NF-κB ( E ), and acetyl-NF-κB ( F ) in CD38 + NK cells from different groups. GAPDH was used as an internal control. CD38 + NK (HC): CD38 + NK cells isolated from the blood of HCs; CD38 + NK (CRC): CD38 + NK cells isolated from the blood of CRC patients; HCs: healthy controls; CRC: colorectal cancer. CD38 mAb: anti-CD38 monoclonal antibody; C-IgG: control IgG. **: P < 0.01 and ***: P < 0.001.

Article Snippet: CD38 + NK cells were incubated with a CD38 mAb (Proteintech) at final concentrations of 0, 1, 5 and 10 μg/ml at 37 °C in 5% CO 2 .

Techniques: Expressing, Gene Expression, Real-time Polymerase Chain Reaction, Western Blot, Quantitation Assay, Control, Isolation

Journal: Scientific Reports

Article Title: CD38 modulates cytokine secretion by NK cells through the Sirt1/NF-κB pathway, suppressing immune surveillance in colorectal cancer

doi: 10.1038/s41598-024-79008-8

Figure Lengend Snippet: The effect of CD38-mediated Sirt1/NF-κB signaling on cytokine secretion and the resultant modulation of CD4 + T-cell differentiation. Expression levels of NF-κB and acetyl-NF-κB proteins in CD38 + NK cells treated with RSV (Sirt1 activator) or DMSO ( A ) and quantitation for NF-κB ( B ) and acetyl-NF-κB ( C ). Expression levels of NF-κB and acetyl-NF-κB proteins in CD38 + NK cells treated with NAM (Sirt1 inhibitor) or DMSO ( D ) and quantitation for NF-κB ( E ) and acetyl-NF-κB ( F ). ( G ) Acetyl-NF-κB expression was compared with NF-κB expression. ( H ) Cytokine levels in the culture supernatant of CD38 + NK cells treated with PDTC (NF-κB inhibitor) or PBS. (I). CD4 + T cells were cocultured with CD38 + NK cells pretreated with PDTC or PBS. ( J ). Proportions of Tregs, Th1, Th2, and Th17 cells and the ratios of Th17/Treg and Th1/Th2 cells among CD4 + T cells cocultured with CD38 + NK cells that were pretreated with PDTC or PBS. CD38 + NK (HC): CD38 + NK cells isolated from the blood of healthy controls; CD38 + NK (CRC): CD38 + NK cells isolated from the blood of CRC patients; CRC cells: colorectal cancer; RSV: resveratrol; NAM: nicotinamide; DMSO: dimethyl sulfoxide. ns: not statistically significant; *: P < 0.05, **: P < 0.01 and ***: P < 0.001.

Article Snippet: CD38 + NK cells were incubated with a CD38 mAb (Proteintech) at final concentrations of 0, 1, 5 and 10 μg/ml at 37 °C in 5% CO 2 .

Techniques: Cell Differentiation, Expressing, Quantitation Assay, Isolation

Journal: Scientific Reports

Article Title: CD38 modulates cytokine secretion by NK cells through the Sirt1/NF-κB pathway, suppressing immune surveillance in colorectal cancer

doi: 10.1038/s41598-024-79008-8

Figure Lengend Snippet: Effects of CD38 + NK cells on macrophage polarization. ( A ) M0 macrophages derived from THP-1 cells were cultured with the culture supernatant of CD38 + NK cells from HCs or CRC patients and pretreated with a CD38 mAb or C-IgG. ( B ) Representative samples from different groups showing the effect of CD38 + NK cell culture supernatant on macrophage polarization. ( C ) Proportions of M1 and M2 macrophages after the treatment of M0 macrophages with the culture supernatant of CD38 + NK cells from different groups. CD38 + NK (HC): CD38 + NK cells isolated from the blood of healthy controls; CD38 + NK (CRC): CD38 + NK cells isolated from the blood of CRC patients; HCs: healthy controls; CRC: colorectal cancer; CD38 mAb: anti-CD38 monoclonal antibody; C-IgG: control IgG. ns: not statistically significant; **: P < 0.01 and ***: P < 0.001.

Article Snippet: CD38 + NK cells were incubated with a CD38 mAb (Proteintech) at final concentrations of 0, 1, 5 and 10 μg/ml at 37 °C in 5% CO 2 .

Techniques: Derivative Assay, Cell Culture, Isolation, Control

Journal: Clinical Cancer Research

Article Title: Preclinical Characterization and Phase I Trial Results of INBRX-109, A Third-Generation, Recombinant, Humanized, Death Receptor 5 Agonist Antibody, in Chondrosarcoma

doi: 10.1158/1078-0432.CCR-23-0974

Figure Lengend Snippet: INBRX-109 characterization. A, Schematic representation of INBRX-109 structure. B, Representative binding curve of INBRX-109 on ExpiCHO-S transfected with full-length human DR5; untransfected ExpiCHO-S cells served as a negative control. Apparent affinity of the observed binding interaction was determined using a One Site total analysis. C, INBRX-109 competition with TRAIL for binding to DR5 expressed by ExpiCHO-S cells. Detection of a constant concentration of TRAIL (3.5 nmol/L) in the presence of increasing concentrations of INBRX-109 is shown. D, ADCC capability of INBRX-109 was evaluated in the Promega Jurkat CD16a (V158) ADCC reporter assay using DR5-transfected ExpiCHO-S cells as targets and untransfected cells as negative controls. Daratumumab, the target of which is expressed on Jurkat cells, serves as a positive control. E, The ability of INBRX-109 to bind the complement component C1q contained within normal human serum was measured by ELISA. Daratumumab is used as a positive control. Abbreviations: Ab, antibody; ADCC, antibody-dependent cellular cytotoxicity; CDC, complement-dependent cytotoxicity; CHO, Chinese hamster ovary cells; DR5, death receptor 5; Fc, crystallizable fragment; K d , equilibrium dissociation constant; sdAb, single-domain antibody; UT, untransfected.

Article Snippet: Jurkat CD16a-V158 ADCC cells Bioassay Effector Cells were cocultured with DR5-ExpiCHO-S or ExpiCHO-S cells and then incubated with INBRX-109 or daratumumab (MedChemExpress, Catalog No. HY-P9915) along with Z-VAD-FMK (50 μmol/L).

Techniques: Binding Assay, Transfection, Negative Control, Concentration Assay, Reporter Assay, Positive Control, Enzyme-linked Immunosorbent Assay

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: IRF4 Is a Critical Gene in Retinoic Acid-Mediated Plasma Cell Formation and Is Deregulated in Common Variable Immunodeficiency-Derived B Cells.

doi: 10.4049/jimmunol.1500250

Figure Lengend Snippet: FIGURE 5. RA induces transient induction of AID and sustained induction of Blimp1 and XBP1s. (A and B) CD19+ B cells (4.0 3 105/ml) were isolated from buffy coats and cultured with various combinations of CpG-ODNs (1 mg/ml), anti-RP105 (1 mg/ml), and RA (100 nM) for the indicated time points. (A) The cells were subjected to Western blot analysis as described in Materials and Methods. The blot represents one representative ex- periment of three. (B) The protein levels were quantitated and normalized to calnexin, and the ratios are displayed. The data represent the mean values 6 SEM of three independent experiments. (C and D) CD19+ B cells (4.0 3 105cells/ml) were isolated from buffy coats and stimulated with of CpG-ODNs (1 mg/ml), anti-RP105 (1 mg/ml), and RA (100 nM) for 5 d. The cells were sorted based on their expression of CD20 and CD38 using a FACSDiva (BD Biosciences) and subjected to Western blot analysis as described in Materials and Methods. The blot shows one representative experiment (C) of two. The three fractions that were obtained in both experiments were quantitated and normalized to calnexin, and the ratios are displayed as mean values 6 SEM (D). aRP, anti-RP105.

Article Snippet: PE-conjugated anti-human CD38 (clone HIT2) and anti-human PerCP-conjugated mouse IgG1 were from BD Biosciences (Franklin Lakes, NJ), and anti-human PerCP-conjugated anti-CD20 (clone LT20) and PE-conjugated mouse IgG1 (clone IS5-21F5) were purchased from Miltenyi Biotec (Bergisch Gladbach, Germany).

Techniques: Isolation, Cell Culture, Western Blot, Expressing

Journal: Frontiers in Immunology

Article Title: Hyper-Expression of PD-1 Is Associated with the Levels of Exhausted and Dysfunctional Phenotypes of Circulating CD161 ++ TCR iVα7.2 + Mucosal-Associated Invariant T Cells in Chronic Hepatitis B Virus Infection

doi: 10.3389/fimmu.2018.00472

Figure Lengend Snippet: Frequencies and levels of immune senescence, exhaustion, and activation markers expressed on CD4 + and CD8 + T-cells between chronic hepatitis B virus (HBV)-infected patients with (circle) and without (square) HBV-DNAemia and healthy control (HC) (triangle). (A) The gating strategy to identify (B) expression levels of CD57, PD-1, TIM-3, CTLA-4 CD38, and HLA-DR on CD4 + and CD8 + T-cells. Levels of surface markers were compared across the three patient groups and post hoc Mann–Whitney U tests were then performed for those biomarkers with a Kruskal–Wallis test P value of <0.05 (* P < 0.05, **<0.01, ***<0.001, and ****<0.0001). P -values remained significant after Benjamini–Hochberg correction of multiple comparisons (marked in red *).

Article Snippet: The second panel was performed with FITC-conjugated anti-HLA-DR, PE-conjugated anti-CD38, PerCP-Cy5.5-conjugated anti-CD3, PE-conjugated anti-TCR-Va7.2-Vio770 (MiltenyiBiotec), APC-conjugated anti-CD161, APC-H7-conjugated anti-CD8, V500-conjugated CD4, and BV-421-conjugated anti-CTLA-4.

Techniques: Activation Assay, Infection, Expressing, MANN-WHITNEY

Journal: Frontiers in Immunology

Article Title: Hyper-Expression of PD-1 Is Associated with the Levels of Exhausted and Dysfunctional Phenotypes of Circulating CD161 ++ TCR iVα7.2 + Mucosal-Associated Invariant T Cells in Chronic Hepatitis B Virus Infection

doi: 10.3389/fimmu.2018.00472

Figure Lengend Snippet: Frequencies and levels of immune senescence, exhaustion, and activation markers expressed on mucosal-associated invariant T (MAIT) cells between chronic hepatitis B virus (HBV)-infected patients with (G1, circle) and without (G2, square) HBV-DNAemia and healthy control (HC) (G3, triangle). (A) The gating strategy to identify CD3 + TCR Vα7.2 + MAIT cells and Vα7.2 + CD161 − T cells. (B) Comparison of T cell receptor (TCR) Vα7.2 + cells frequencies across the three groups comparison of percentage. (C) Expression levels [mean fluorescence intensity (MFI)] (D) of CD57, PD-1, TIM-3, CTLA-4 CD38 and HLA-DR on TCR Vα7.2 + MAIT cells between the three patient groups. Levels of surface markers were compared across the three patient groups and post hoc Mann–Whitney U tests were then performed for those biomarkers with a Kruskal–Wallis test P value of <0.05 (* P < 0.05, **<0.01, ***<0.001, and ****<0.0001). P -values remained significant after Benjamini–Hochberg correction of multiple comparisons (marked in red *).

Article Snippet: The second panel was performed with FITC-conjugated anti-HLA-DR, PE-conjugated anti-CD38, PerCP-Cy5.5-conjugated anti-CD3, PE-conjugated anti-TCR-Va7.2-Vio770 (MiltenyiBiotec), APC-conjugated anti-CD161, APC-H7-conjugated anti-CD8, V500-conjugated CD4, and BV-421-conjugated anti-CTLA-4.

Techniques: Activation Assay, Infection, Expressing, Fluorescence, MANN-WHITNEY

Journal: Frontiers in Immunology

Article Title: Hyper-Expression of PD-1 Is Associated with the Levels of Exhausted and Dysfunctional Phenotypes of Circulating CD161 ++ TCR iVα7.2 + Mucosal-Associated Invariant T Cells in Chronic Hepatitis B Virus Infection

doi: 10.3389/fimmu.2018.00472

Figure Lengend Snippet: (A) Comparison of serum alanine transaminase (ALT) levels between chronic hepatitis B virus (HBV)-infected patients with HBV-DNAemia (circle) and without HBV-DNAemia (square). (B) Spearman correlation between serum ALT levels and frequencies of T cell receptor (TCR) Vα7.2 + CD161 + mucosal-associated invariant T (MAIT) cells. (C) Spearman correlation between frequencies and expression levels of CD57, PD-1, TIM-3, CTLA-4 CD38, and HLA-DR with serum level of ALT. The bar represents the strength of association ( r values) where red bar represents significant positive correlation, blue bar represent significant negative association and black bar represents P value > 0.05 (non-significant association) (<0.05, **<0.01, ***<0.001, and ****<0.0001).

Article Snippet: The second panel was performed with FITC-conjugated anti-HLA-DR, PE-conjugated anti-CD38, PerCP-Cy5.5-conjugated anti-CD3, PE-conjugated anti-TCR-Va7.2-Vio770 (MiltenyiBiotec), APC-conjugated anti-CD161, APC-H7-conjugated anti-CD8, V500-conjugated CD4, and BV-421-conjugated anti-CTLA-4.

Techniques: Infection, Expressing

Journal: Signal Transduction and Targeted Therapy

Article Title: Myeloid but not hepatocytic CD38 is a key driver for hepatic ischemia/reperfusion injury

doi: 10.1038/s41392-025-02233-8

Figure Lengend Snippet: CD38 deficiency alleviated hepatic ischemia-reperfusion injury (HIRI) in mice. a CD38 expressions were determined by Western blotting and quantitative analysis in liver tissues after ischemia/reperfusion (I/R) injury. b The mRNA expressions of IL1B and IL-6 in liver tissues after I/R injury. c Representative fluorescence images of IL-1β/F4/80 and IL-1β/ASGR1 were taken from CD38 KO mice subjected to HIRI, respectively. d , e Serum ALT and AST were measured in CD38 KO and CD38 MKO mice subjected to HIRI. f , g Representative H&E staining images and the quantitative analysis of liver ischemic necrosis were taken from CD38 KO and CD38 MKO mice subjected to HIRI, respectively. Data are shown as means ± SEM, * p < 0.05, ** p < 0.01 and *** p < 0.001, n = 3–9 per group

Article Snippet: CD38 (R&D), GAPDH (KangChen), SOD2 (CST), NOX2 (Abcam), IL-1β (CST), PPARγ (Santa Cruz Biotechnology), NLRP3 (CST), IL-18 (Abmart), GSDMD-N (Abmart), SIRT3 (Sigma-Aldrich), SIRT1 (Sigma-Aldrich), p53 (Abcam), Acetyl-p53 (Abcam) antibodies were diluted 1:400/1000, respectively.

Techniques: Western Blot, Fluorescence, Staining

Journal: Signal Transduction and Targeted Therapy

Article Title: Myeloid but not hepatocytic CD38 is a key driver for hepatic ischemia/reperfusion injury

doi: 10.1038/s41392-025-02233-8

Figure Lengend Snippet: Global CD38 deficiency inhibited oxidative stress and inflammation during HIRI. a Representative DHE staining images were taken from CD38 KO and CD38 fl/fl mice subjected to HIRI, respectively. b MDA contents were quantitatively measured in liver tissues from CD38 KO and CD38 fl/fl mice after I/R injury. c Serum TNF-α and IL-1β were detected by Elisa in CD38 KO and CD38 fl/fl mice after HIRI. d The protein expressions and the quantitative analysis of NOX2 and SOD2 were determined by western blot in liver tissues from CD38 KO and CD38 fl/fl mice after HIRI. e The mRNA expressions of TNF-α, IL1B, TGF-β, CD206, and PPARγ were determined by QPCR in CD38 KO and CD38 fl/fl mice after HIRI, respectively. f , g The expressions and the quantitative analysis of IL-1β and PPARγ were detected by Western blot in liver tissues in CD38 KO and CD38 fl/fl mice after HIRI, respectively. Data are shown as means ± SEM, * p < 0.05, ** p < 0.01 and *** p < 0.001, n = 3–8 per group

Article Snippet: CD38 (R&D), GAPDH (KangChen), SOD2 (CST), NOX2 (Abcam), IL-1β (CST), PPARγ (Santa Cruz Biotechnology), NLRP3 (CST), IL-18 (Abmart), GSDMD-N (Abmart), SIRT3 (Sigma-Aldrich), SIRT1 (Sigma-Aldrich), p53 (Abcam), Acetyl-p53 (Abcam) antibodies were diluted 1:400/1000, respectively.

Techniques: Staining, Enzyme-linked Immunosorbent Assay, Western Blot

Journal: Signal Transduction and Targeted Therapy

Article Title: Myeloid but not hepatocytic CD38 is a key driver for hepatic ischemia/reperfusion injury

doi: 10.1038/s41392-025-02233-8

Figure Lengend Snippet: Myeloid CD38 deficiency reduced hepatic ischemia-reperfusion-triggered inflammation. a – c The mRNA expressions of IL1B, TNF-α, iNOS, IL-10, ARG1, TGF-β and PPARγ were analyzed by QPCR in CD38 MKO and CD38 fl/fl mice after HIRI. d The expressions and the quantitative analysis of PPARγ were detected by Western blot in liver tissues of CD38 MKO and CD38 fl/fl mice after HIRI. e MDA contents were measured in liver tissues of CD38 LKO and CD38 fl/fl mice after HIRI. f The mRNA expressions of TNF-α and IL1B were determined by QPCR in liver tissues of CD38 LKO and CD38 fl/fl mice after HIRI. Data are shown as means ± SEM, * p < 0.05, ** p < 0.01 and *** p < 0.001, n = 3 ~ 9 per group

Article Snippet: CD38 (R&D), GAPDH (KangChen), SOD2 (CST), NOX2 (Abcam), IL-1β (CST), PPARγ (Santa Cruz Biotechnology), NLRP3 (CST), IL-18 (Abmart), GSDMD-N (Abmart), SIRT3 (Sigma-Aldrich), SIRT1 (Sigma-Aldrich), p53 (Abcam), Acetyl-p53 (Abcam) antibodies were diluted 1:400/1000, respectively.

Techniques: Western Blot

Journal: Signal Transduction and Targeted Therapy

Article Title: Myeloid but not hepatocytic CD38 is a key driver for hepatic ischemia/reperfusion injury

doi: 10.1038/s41392-025-02233-8

Figure Lengend Snippet: Global and myeloid deletion of CD38 ameliorated HIRI-induced pyroptosis in vivo. The protein expressions and the quantitative analysis of NLRP3, IL-18 and GSDMD-N and the mRNA expressions of NLRP3, Caspase-1 and IL-18 were examined by Western blot ( a ) and QPCR ( b ) in liver tissues of CD38 KO and CD38 fl/fl mice after HIRI, respectively. The protein expressions and the quantitative analysis of NLRP3, GSDMD-N and IL-18, and the mRNA expressions of NLRP3, Caspase-1 and IL-18 were determined by Western blot ( c ) and QPCR ( d ) in liver tissues from CD38 MKO and CD38 fl/fl mice after HIRI. Data are shown as means ± SEM, * p < 0.05, ** p < 0.01and *** p < 0.001, n = 3–6 per group

Article Snippet: CD38 (R&D), GAPDH (KangChen), SOD2 (CST), NOX2 (Abcam), IL-1β (CST), PPARγ (Santa Cruz Biotechnology), NLRP3 (CST), IL-18 (Abmart), GSDMD-N (Abmart), SIRT3 (Sigma-Aldrich), SIRT1 (Sigma-Aldrich), p53 (Abcam), Acetyl-p53 (Abcam) antibodies were diluted 1:400/1000, respectively.

Techniques: In Vivo, Western Blot

Journal: Signal Transduction and Targeted Therapy

Article Title: Myeloid but not hepatocytic CD38 is a key driver for hepatic ischemia/reperfusion injury

doi: 10.1038/s41392-025-02233-8

Figure Lengend Snippet: Myeloid CD38 deficiency alleviated hepatocytic hypoxia/reoxygenation injury in a co-culture condition in vitro. a Representative flow cytometry plots of PI staining, along with the quantitative analysis, for hepatocytes from CD38 fl/fl mice co-cultured with BMDMs from CD38 KO mice after H/R injury. MDA contents ( b ), LDH activities ( c ), the mRNA expressions of IL-1B ( d ), TNF-α ( d ), NLRP3 ( e ) and IL-18 ( e ), and the protein expressions and the quantitative analysis ( f ) of NLRP3, IL-18 and PPARγ were examined in primary hepatocytes from CD38 fl/fl mice co-cultured with BMDMs from CD38 KO mice after H/R injury. Data are shown as means ± SEM, * p < 0.05, ** p < 0.01 and *** p < 0.001, n = 3–6 per group

Article Snippet: CD38 (R&D), GAPDH (KangChen), SOD2 (CST), NOX2 (Abcam), IL-1β (CST), PPARγ (Santa Cruz Biotechnology), NLRP3 (CST), IL-18 (Abmart), GSDMD-N (Abmart), SIRT3 (Sigma-Aldrich), SIRT1 (Sigma-Aldrich), p53 (Abcam), Acetyl-p53 (Abcam) antibodies were diluted 1:400/1000, respectively.

Techniques: Co-Culture Assay, In Vitro, Flow Cytometry, Staining, Cell Culture

Journal: Signal Transduction and Targeted Therapy

Article Title: Myeloid but not hepatocytic CD38 is a key driver for hepatic ischemia/reperfusion injury

doi: 10.1038/s41392-025-02233-8

Figure Lengend Snippet: Global and myeloid deletion of CD38 upregulated SIRT1-p53 signaling pathway. The expressions and the quantitative analysis of SIRT1 ( a, b ), SIRT3 ( a, b ), p53 ( c, d ), and AC-p53 ( c, d ) were determined by Western blot in liver tissues from CD38 KO , CD38 MKO , and CD38 fl/fl mice after HIRI. Data are shown as means ± SEM, * p < 0.05, ** p < 0.01 and *** p < 0.001, n = 3 per group

Article Snippet: CD38 (R&D), GAPDH (KangChen), SOD2 (CST), NOX2 (Abcam), IL-1β (CST), PPARγ (Santa Cruz Biotechnology), NLRP3 (CST), IL-18 (Abmart), GSDMD-N (Abmart), SIRT3 (Sigma-Aldrich), SIRT1 (Sigma-Aldrich), p53 (Abcam), Acetyl-p53 (Abcam) antibodies were diluted 1:400/1000, respectively.

Techniques: Western Blot

Journal: Signal Transduction and Targeted Therapy

Article Title: Myeloid but not hepatocytic CD38 is a key driver for hepatic ischemia/reperfusion injury

doi: 10.1038/s41392-025-02233-8

Figure Lengend Snippet: Myeloid CD38 deficiency promoted macrophage M2 polarization through activating NAD + /SIRT1 pathway in vitro. The mRNA expressions of IL1B ( a ), CD206 ( a ), and the protein expressions and the quantitative analysis of SIRT1 ( b ) and SIRT3 ( b ) were determined by QPCR and Western blot in BMDMs from CD38 fl/fl and CD38 KO mice after LPS stimulation, respectively. c The mRNA expressions of IL1B and IL-6 were determined by QPCR in BMDMs from CD38 fl/fl mice with the stimulation of hepatocytes-derived conditioned media after hypoxia/reoxygenation injury (HCMHR). The protein expressions and the quantitative analysis of SIRT1/SIRT3 ( d ), the NAD + contents ( e ) and the infiltrations of the type 2 macrophages (CD206/Il-10, f, g ), and type 1 macrophages (CD86/iNOS, f, g ) were determined in BMDMs from CD38 KO and CD38 fl/fl mice with HCMHR stimulation. Data are shown as means ± SEM, * p < 0.05, ** p < 0.01 and *** p < 0.001, n = 3–6 per group

Article Snippet: CD38 (R&D), GAPDH (KangChen), SOD2 (CST), NOX2 (Abcam), IL-1β (CST), PPARγ (Santa Cruz Biotechnology), NLRP3 (CST), IL-18 (Abmart), GSDMD-N (Abmart), SIRT3 (Sigma-Aldrich), SIRT1 (Sigma-Aldrich), p53 (Abcam), Acetyl-p53 (Abcam) antibodies were diluted 1:400/1000, respectively.

Techniques: In Vitro, Western Blot, Derivative Assay

Journal: Signal Transduction and Targeted Therapy

Article Title: Myeloid but not hepatocytic CD38 is a key driver for hepatic ischemia/reperfusion injury

doi: 10.1038/s41392-025-02233-8

Figure Lengend Snippet: Myeloid CD38 deficiency facilitated macrophage M2-type polarization through activating SIRT1-p53 and SIRT1-PPARγ pathway in vitro . The mRNA expressions of TNF-α ( a ), IL1B ( b ), IL-10 ( c ) and CD206 ( d ) were determined by QPCR in BMDMs from CD38 KO and CD38 fl/fl mice with the stimulation of the hepatocytes-derived conditioned media after hypoxia/reoxygenation injury (HCMHR) in the pretreatment of Compound C (AMPK inhibitor), PFT-α (P53 inhibitor) and T0070907(PPARγ inhibitor), respectively. Data are shown as means ± SEM, * p < 0.05, ** p < 0.01 and *** p < 0.001, n = 3 per group. e The mechanism of myeloid-specific deletion of CD38 (CD38 MKO ) protecting against hepatic ischemia/reperfusion injury (HIRI): CD38 MKO elevates the intracellular NAD + levels in macrophages, and in turn, alleviates hepatic ischemia/reperfusion injury (HIRI)-induced inflammation and pyroptosis via activating SIRT1 signaling pathways in macrophages. On the one hand, SIRT1 promotes myeloid monocytes toward macrophage type 2 polarization through activating PPARγ signaling pathway in macrophages and inhibits monocytes toward macrophage type 1 polarization via activating p53 signaling, reducing HIRI-induced inflammation. On the other hand, SIRT1 also suppresses the release of pro-inflammatory factors such as IL1-β and IL18 through inactivating the canonical inflammasome-pyroptosis pathway of NLRP3-mediated caspase-1/GSDMD processing in macrophages. Image created with BioRender.com, with permission (agreement number: FG27ZL6X5O; citation to use: https://www.biorender.com/j12e628 )

Article Snippet: CD38 (R&D), GAPDH (KangChen), SOD2 (CST), NOX2 (Abcam), IL-1β (CST), PPARγ (Santa Cruz Biotechnology), NLRP3 (CST), IL-18 (Abmart), GSDMD-N (Abmart), SIRT3 (Sigma-Aldrich), SIRT1 (Sigma-Aldrich), p53 (Abcam), Acetyl-p53 (Abcam) antibodies were diluted 1:400/1000, respectively.

Techniques: In Vitro, Derivative Assay, Protein-Protein interactions