Journal: EMBO Molecular Medicine

Article Title: Mitochondrial damage drives T-cell immunometabolic paralysis after major surgery

doi: 10.1038/s44321-025-00324-1

Figure Lengend Snippet: ( A ) Scheme of the study flow and time points of blood withdrawal during the course of major surgery. ( B ) mRNA expression of IFNG (left), GZMB (middle) and PRF1 (right) in CTL (+CD3/CD28 dynabeads, stimulated for 18 h) relative to endogenous controls, n = 9/10/10 individual patients. ( C ) Flow cytometric analysis (FCA) of intracellular IFN-γ staining and quantification in CTL (+CD3/CD28 dynabeads, stimulated for 18 h; re-stimulation with PMA/Ionomycin for 4 h), n = 9 individual patients. ( D ) Quantification of IFN-γ secretion by CTL (+CD3/CD28 dynabeads, stimulated for 18 h), n = 9 individual patients. ( E ) Representative impedance plot (left) and quantification of relative CTL cytotoxicity (right) using ECIS, n = 7 individual patients. ( F ) FCA of CD38 staining and quantification in CTL, minimum = 28.6/32.9, 25%-percentile = 46.73/55.65, median = 59.65/67.05, 75%-percentile = 69.10/75.75, maximum = 75.10/80.60 (T1/T2), n = 10 individual patients. ( G ) Quantification of GZMB (left) and PRF1 (right) secretion by CTL (+CD3/CD28 dynabeads, stimulated for 18 h) as measured by ELISA, n = 11/12/6 (GZMB) and 13/13/6 (PRF1) individual patients. ( H ) FCA of intracellular GZMB staining and quantification in CTL (+CD3/CD28 dynabeads, stimulated for 18 h; re-stimulation with PMA/Ionomycin for 4 h), n = 9 individual patients. ( I ) FCA of intracellular PRF1 staining and quantification in CTL, n = 9 individual patients. ( J ) FCA of CD107a staining and quantification in CTL (+CD3/CD28 dynabeads, stimulated for 18 h; re-stimulation with PMA/Ionomycin for 4 h), n = 6 individual patients. If not stated otherwise, data are represented as mean ± SEM. P values as indicated, paired t -test or Wilcoxon matched-pairs signed-rank test, as appropriate. .

Article Snippet: CD38 APC , Miltenyi , 130-113-991.

Techniques: Expressing, Staining, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Immunology

Article Title: Unveiling spatial complexity in solid tumor immune microenvironments through multiplexed imaging

doi: 10.3389/fimmu.2024.1383932

Figure Lengend Snippet: Immunophenotyping panel for multiplexed tissue imaging of cancer.

Article Snippet: CD38 , REA671 , 50 , 130-117-717 , PE (APC) , Miltenyi Biotec.

Techniques: Imaging

Journal: Frontiers in Immunology

Article Title: Unveiling spatial complexity in solid tumor immune microenvironments through multiplexed imaging

doi: 10.3389/fimmu.2024.1383932

Figure Lengend Snippet: Deep spatial profiling of human palatine tonsil tissues. (A) Hematoxylin and eosin (H&E) staining after 92 MICS cycles, including the marked epithelium, germinal center (GC), and T cell zone of the lymphoid follicle. MICS DAPI and stroma staining depicting the composition and structure of the tonsil. Markers: Collagen III, collagen IV, fibronectin (all extracellular matrix (ECM), cytokeratin (epithelium), podoplanin (lymphatic vessels), CD105/SM Actin (blood vessels). (B) Immune cell content of a human palatine tonsil comprising T cells (CD3), B cells (CD19/CD20), plasma cells (PCs) (CD38/CD138), NK cells (CD56), granulocytes (CD15/CD66b), mast cells (CD117), macrophages (MΦ) (CD163/CD169/CD206), myeloid dendritic cells (mDCs) (CD11c), and plasmacytoid dendritic cells (pDCs) (CD123). (C) Detailed view on the T cell zone, mainly composed of CD4 + helper T cells (T h ) and CD8 + cytotoxic T cells (T c ), mDCs (CD11c), and PCs (CD38/CD138). (D) Detailed view on the GC-mantle zone border, showing different B cells (CD11b, CD21, CD22), mDCs (CD11c), and PCs (CD38/CD138). (E) Cell annotations of three different tonsil samples plus respective bar graphs of gated cell populations, comparing the cell content between the three tonsil samples. Depicted markers and annotated cell types as indicated by the color code. ROI sizes: 976 x 640 µm, zoomed-in subregions in (C, D) : 334 µm x 219 µm. Scale bar: 100 µm.

Article Snippet: CD38 , REA671 , 50 , 130-117-717 , PE (APC) , Miltenyi Biotec.

Techniques: Staining, Clinical Proteomics

Journal: Cancers

Article Title: Neoantigen Dendritic Cell Vaccination Combined with Anti-CD38 and CpG Elicits Anti-Tumor Immunity against the Immune Checkpoint Therapy-Resistant Murine Lung Cancer Cell Line LLC1

doi: 10.3390/cancers13215508

Figure Lengend Snippet: The adenosine pathway in the LLC1 tumor microenvironment. ( A ) RNA was extracted from ASB-XIV and LLC1 tumor cells and subjected to RNA-Seq analysis. The mRNA expressions of immunosuppressive molecules in these two cell lines were shown. ( B ) The expressions of CD38, CD39, and CD73 on LLC1 were examined by flow cytometry in vitro. ( C ) C57BL/6 mice ( n = 5) received a subcutaneous injection of 5 × 10 5 LLC1. The combination of 2 mg/Kg A2A and 1 mg/Kg A2B adenosine receptor antagonists (SCH 58261 and PSB 1115) was administered on days 7, 9, 11, 13, 15, 17, and 19. ( D ) LLC1 tumor growth was compared in mice ( n = 8) that received 30μg CpG monotherapy, L82 LP-pulsed DC vaccination (1 × 10 6 ), and CpG, or the triple combination of DC vaccination, CpG, and A2R antagonists. DC vaccines were prophylactically administered twice at a 2-week interval.

Article Snippet: Monoclonal antibodies (mAbs) specific for PD-1 (RMP1-14), CTLA-4 (9H10), CD38 (NIMR5), and CD16/32 (2.4G2) were purchased from BioXcell (Lebanon, NH, USA).

Techniques: RNA Sequencing Assay, Flow Cytometry, In Vitro, Injection

Journal: Cancers

Article Title: Neoantigen Dendritic Cell Vaccination Combined with Anti-CD38 and CpG Elicits Anti-Tumor Immunity against the Immune Checkpoint Therapy-Resistant Murine Lung Cancer Cell Line LLC1

doi: 10.3390/cancers13215508

Figure Lengend Snippet: The triple combination of DC vaccination, CpG and anti-CD38 mAb treatment inhibited LLC1 tumor growth. ( A ) C57BL/6 mice were divided into five groups (five mice per group): (1) 30μg CpG treatment; (2) CpG plus 250μg anti-CD38 mAb; (3) 1×10 6 L82 LP-pulsed DC vaccination with CpG; (4) the triple combination of L82 LP-pulsed DC vaccination, CpG, and anti-CD38 mAb; and (5) the quadruple combination of DC, CpG, anti-CD38 mAb, and 200μg of anti-PD-1 mAb. LLC1 cells (5 × 10 5 ) were subcutaneously inoculated and tumor growth was monitored. ( B ) On day 14, tumor-infiltrating cells were extracted from these mice ( n = 5) and subjected to flow cytometry. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Monoclonal antibodies (mAbs) specific for PD-1 (RMP1-14), CTLA-4 (9H10), CD38 (NIMR5), and CD16/32 (2.4G2) were purchased from BioXcell (Lebanon, NH, USA).

Techniques: Flow Cytometry

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Reduced ALDH1A1 expression in multiple myeloma cells increases resistance to daratumumab via downregulation of retinoic acid

doi: 10.1007/s00018-025-05891-7

Figure Lengend Snippet: ALDH1A1 increased after conventional chemotherapy regimen Vrd or Krd. A and B. GSE276561 and GSE47552 datasets revealed ALDH1A1 in MM was lower than healthy control; These GEO data were analyzed by on line tool GEO2R using Benjamini & Hochberg (False discovery rate) method; Each y axis label in the graph represents the expression measurement extracted from the TPM normalized expression counts (for RNA-seq), or the Value column of the original submitter-supplied Sample record (for microarrays). All the y axis label represents normalized mRNA expression level. C. mRNA levels of ALDH1A1 in NDMM samples are significantly lower than healthy control, but remarkably higher in RRMM samples than NDMM patients which are collected in our hospital; Relative ALDH1A1 levels is relative to GAPDH. D. mRNA levels of CD38 in NDMM patients are significantly higher than healthy control which are collected in our hospital; Relative CD38 levels is relative to GAPDH. E. ALDH1A1 mRNA increased after Vrd or Krd treatment; Relative ALDH1A1 levels is relative to GAPDH. F . western blot confirmed ALDH1A1 protein increased after Vrd or Krd treatment. **, p < 0.01, ***, p < 0.001. C-E, Data was analyzed by Mann-Whitnay test HC: healthy control; Vrd: Bortezomib + Lenalidomide + Dexamethasone; KRd: Carfilzomib + Lenalidomide + Dexamethasone; MM: multiple myeloma; NDMM: newly diagnosed multiple myeloma; SMM: smoldering multiple myeloma; MGUS: monoclonal gammopathy of undetermined significance

Article Snippet: The positive CD38 cells were labeled by APC anti-human CD38 antibody (E-AB-F1058E, Elabscience) or APC anti-mouse CD38 antibody (E-AB-F1193E, Elabscience) and analyzed by FACSCantoTM II flow cytometer.

Techniques: Control, Expressing, RNA Sequencing, Western Blot

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Reduced ALDH1A1 expression in multiple myeloma cells increases resistance to daratumumab via downregulation of retinoic acid

doi: 10.1007/s00018-025-05891-7

Figure Lengend Snippet: ALDH1A1 down-regulated in the nrRRMM patients received Dara-Rd treatment. A and B. rRRMM patients had higher ALDH1A expression compared with nrRRMM after Dara-Rd treatment analyzed by RT-qPCR and western blot; C and D . the CD38 expression (mRNA and protein) decreased in Dara-treated patients in nrRRMM patients compared with rRRMM patients; E . the mRNA expression of ALDH1A1 after Dara-based treatment was positively correlated with the CD38 mRNA expression. **, p < 0.01 A-D, data was analyzed by student t test. E, data was analyzed by simple linear regression nrRRMM: non-responder relapsed/refractory multiple myeloma; rRRMM: responder relapsed/refractory multiple myeloma; MFI: median fluorescence intensity; Dara-Rd: daratumumab + lenalidomide + dexamethasone; Dara: daratumumab

Article Snippet: The positive CD38 cells were labeled by APC anti-human CD38 antibody (E-AB-F1058E, Elabscience) or APC anti-mouse CD38 antibody (E-AB-F1193E, Elabscience) and analyzed by FACSCantoTM II flow cytometer.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Fluorescence

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Reduced ALDH1A1 expression in multiple myeloma cells increases resistance to daratumumab via downregulation of retinoic acid

doi: 10.1007/s00018-025-05891-7

Figure Lengend Snippet: ALDH1A1 directly regulates CD38 expression in H929 cells. A . the KD efficiency of different sh-ALDH1A1 sequences; B . Knock down ALDH1A1 decreased CD38 genes expression; C. Re-introducing ALDH1A1-OE plasmid restored downregulation of ALDH1A1 mRNA levels in ALDH1A1-KD cells; D . Re-introducing ALDH1A1 into ALDH1A1-KD cells significantly upregulate the expression of CD38; E . cell-surface expression of CD38 decreased in ALDH1A1-KD cells but restored by re-introducing ALDH1A1-OE plasmid determined by flow cytometry analysis. **, p < 0.01, ***, p < 0.001 Student t test was performed in all datasets AlDH1A1-KD: ALDH1A1 knockdown; Ctrl: control; shALDH1A1: short hairpin RNA for ALDH1A1; KD-OE: over expression of ALDH1A1 in KD cells

Article Snippet: The positive CD38 cells were labeled by APC anti-human CD38 antibody (E-AB-F1058E, Elabscience) or APC anti-mouse CD38 antibody (E-AB-F1193E, Elabscience) and analyzed by FACSCantoTM II flow cytometer.

Techniques: Expressing, Knockdown, Plasmid Preparation, Flow Cytometry, Control, shRNA, Over Expression

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Reduced ALDH1A1 expression in multiple myeloma cells increases resistance to daratumumab via downregulation of retinoic acid

doi: 10.1007/s00018-025-05891-7

Figure Lengend Snippet: ALDH1A1 increased CD38 through activation of RA/RAR via RA production. A . Volcano plots created by differential mRNA (fold change (FC) > 2 or < 0.5, and p < 0.05); B. KEGG analysis in ALDH1A1 KD cells of H929 and 8226 cells revealed that retinoic acid receptor, and retinoid X receptor agonists/antagonists pathways were enriched in both cell line; C. Knock down ALDH1A1caused downregulation of RAR proteins while re-introduced ALDH1A1 into ALDH1A1-KD cells increased their expressions; D . RA salvage restore the ADCC of Dara against MM cells; E . the RAR protein level was significantly increased determined by western blot after RA supplement; (F) mRNA levels of CD38 determined by RT-qPCR; (G) protein levels of CD38 determined by western blot. **, p < 0.01, ***, p < 0.001 C-G, student t test was adopted KD: ALDH1A1 knockdown; Ctrl: control; KD-OE: over expression of ALDH1A1 in KD cells; RA: retinoic acid; RAR: retinoic acid receptor; ADCC: antibody-dependent cellular cytotoxicity

Article Snippet: The positive CD38 cells were labeled by APC anti-human CD38 antibody (E-AB-F1058E, Elabscience) or APC anti-mouse CD38 antibody (E-AB-F1193E, Elabscience) and analyzed by FACSCantoTM II flow cytometer.

Techniques: Activation Assay, Knockdown, Western Blot, Quantitative RT-PCR, Control, Over Expression

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Reduced ALDH1A1 expression in multiple myeloma cells increases resistance to daratumumab via downregulation of retinoic acid

doi: 10.1007/s00018-025-05891-7

Figure Lengend Snippet: ALDH1A1 inhibitor reduces the sensitivity of Dara in vivo through inhibiting the RA/RAR pathway. A and B . compared with vehicle control, Dara cause significantly reduction of xenograft tumors weight and tumor growth; C . inhibitor of ALDH1A 673 A remarkably reduce the protein level of CD38, ALDH1A1 and RAR, and RA complement significantly restore the protein level of CD38 and RAR. *, p < 0.05, **, p < 0.01, ***, p < 0.001 A-B, analysis of variance (ANOVA) was used for multi-group comparisons followed by Tukey’s post hoc test ( N = 6); C, analysis of variance (ANOVA) was used for multi-group comparisons followed by Tukey’s post hoc test ( N = 3) RA: retinoic acid; RAR: retinoic acid receptor

Article Snippet: The positive CD38 cells were labeled by APC anti-human CD38 antibody (E-AB-F1058E, Elabscience) or APC anti-mouse CD38 antibody (E-AB-F1193E, Elabscience) and analyzed by FACSCantoTM II flow cytometer.

Techniques: In Vivo, Control