Journal: International Journal of Molecular Sciences

Article Title: The Impact of Duodenal Mucosal Vulnerability in the Development of Epigastric Pain Syndrome in Functional Dyspepsia

doi: 10.3390/ijms232213947

Figure Lengend Snippet: Infiltration of immune cells in duodenal mucosa. Duodenal biopsy specimens obtained from the EPS and PDS patients, and the healthy controls were stained immunohistochemically for mast cell using tryptase antibody ( a ), eosinophils using eosinophilic MBP antibody ( b ) and Ig-E in plasma cells using Ig-E ( c ). The immunostaining of each antibody was quantified by counting on 3 randomly selected high-power fields for each sample (×400 magnification) ( d , e ). The number of Ig-E-positive cells was counted within the cells that were stained with CD138, comparing with serial sections of each staining ( f ). The immune-positive cells were counted in 3 randomly selected high-power fields on each sample (controls: n = 23, EPS patients: n = 16, PDS patients: n = 11). EPS: epigastric pain syndrome, PDS: postprandial distress syndrome, MBP: major basic protein. * p < 0.05, Tukey–Kramer test, NS: not significant, Bar indicates 20 μm.

Article Snippet: The tissues were treated with a target retrieval solution (Dako, Glostrup, Denmark) in a pressure cooker at 125 °C for 5 min. Eosinophils, mast cells and plasma cells were incubated with mouse antieosinophilic major basic protein (MBP) (1:20; AbD Serotec, Kidlington, UK), rabbit antimast cell tryptase (1:200; Dako) and mouse antihuman CD138 (1:50; Dako) for plasma cells at 4 °C overnight incubation, respectively.

Techniques: Staining, Clinical Proteomics, Immunostaining

Journal: Journal of Vascular Research

Article Title: Rivaroxaban as a Protector of Oxidative Stress-Induced Vascular Endothelial Glycocalyx Damage via the IQGAP1/PAR1-2/PI3K/Akt Pathway

doi: 10.1159/000542419

Figure Lengend Snippet: sCD138 release under oxidative stress and the effect of rivaroxaban in ECs. a sCD138 concentration in the culture supernatant of HUVECs 20 min after 1, 50, 100 m m H 2 O 2 stimulation ( n = 5 for each culture). b ECs viability in 20 min after 1, 50, 100 m m H 2 O 2 stimulation ( n = 4 for each culture). c FX levels measured in HUVEC cell lysates with or without 20 min H 2 O 2 treatment ( n = 2 for each culture). d sCD138 concentration in the 1 m m H 2 O 2 -stimulated HUVEC culture supernatant with or without rivaroxaban pretreatment ( n = 4 for each culture). Bar graph represents averaged data. Values are the mean +/−SE. * p < 0.05 versus control. Significance is assessed using one-way ANOVA, followed by Dunnett’s post hoc analysis. ECs, endothelial cells; sCD138, soluble CD138; HUVECs, human umbilical vein endothelial cells.

Article Snippet: The concentration of soluble syndecan-1 (sCD138) was measured by ELISA (Diaclone 950.640.192, Besançon) at the dilution of 1:1.

Techniques: Concentration Assay, Control

Journal: Journal of Vascular Research

Article Title: Rivaroxaban as a Protector of Oxidative Stress-Induced Vascular Endothelial Glycocalyx Damage via the IQGAP1/PAR1-2/PI3K/Akt Pathway

doi: 10.1159/000542419

Figure Lengend Snippet: Involvement PI3K pathway in the improvement of cell viability and sCD138 shedding by rivaroxaban. a The effect of rivaroxaban in ECs viability in 20 min of 1 m m H 2 O 2 stimulation with or without PI3K inhibitor LY29002 ( n = 5 for each culture). b The effect of rivaroxaban on sCD138 shedding from the cell surface was measured using HUVEC culture supernatant in 20 min of H 2 O 2 stimulation with or without LY29002 ( n = 5 for each culture). Bar graph represents averaged data. Values are the mean +/−SE. * p < 0.05. Significance is assessed using one-way ANOVA, followed by Dunnett’s post hoc analysis. ECs, endothelial cells; PI3K, phosphatidylinositol 3-kinase.

Article Snippet: The concentration of soluble syndecan-1 (sCD138) was measured by ELISA (Diaclone 950.640.192, Besançon) at the dilution of 1:1.

Techniques:

Journal: Cytotherapy

Article Title: Challenges in αCD38-chimeric antigen receptor (CAR)-expressing natural killer (NK) cell-based immunotherapy in multiple myeloma: Harnessing the CD38dim phenotype of cytokine-stimulated NK cells as a strategy to prevent fratricide.

doi: 10.1016/j.jcyt.2023.03.006

Figure Lengend Snippet: Fig. 4. aCD38-CAR-NK-92 cells activate against primary MM cells. (A) Frequency of malignant (CD138+) cell content within the BM-derived mononuclear cell samples of three patients with MM. (B) Flow cytometry plots indicating the CD38 expression of the malignant cells. Intensity of CD38 expression (MFI) is depicted within the plots. (C, D) In vitro acti- vation of aCD38-CAR-NK-92 and NK-92-GFP cells after 4-h contact with BM-derived MNC samples from three patients with MM. Results show (C) degranulation and (D) IFNg release. (E) CD38 surface expression of CD138-selected MM cells from two patients as determined by flow cytometry. Intensity of CD38 expression (MFI) is depicted within the plots. (F) Degranulation of aCD38-CAR-NK-92 and NK-92-GFP cells following 4-h co-culture with primary CD138+-selected MM cells of two patients. (G) Combined results of the degranu- lating capacity of aCD38-CAR-NK-92 compared with NK-92-GFP cells after contact with primary CD138+-selected or CD138+-unselected (MNC) MM cell samples.

Article Snippet: Magnetic separation of the malignant CD138+ fraction from the MNC samples was performed using MACSprep Multiple Myeloma CD138 MicroBeads (Miltenyi Biotec, San Diego, CA, USA) where indicated.

Techniques: Derivative Assay, Flow Cytometry, Expressing, In Vitro, Cytometry, Co-Culture Assay

Journal: Oncogene

Article Title: LAMP5 modulates IRF4 stability and nuclear transport: a critical mechanism in myeloma progression and therapy

doi: 10.1038/s41388-025-03513-x

Figure Lengend Snippet: A , B LAMP5 mRNA expression in myeloma and other indicated cancer cell lines or other tissues, according to the Cancer Cell Line Encyclopedia (CCLE) dataset. C LAMP5 mRNA levels in plasma cells from myeloma patients ( n = 559) compared to normal plasma cells from healthy donors ( n = 22) (GEO: GSE2658 and GSE5900 ) (left). LAMP5 mRNA levels in plasma cells from myeloma patients ( n = 73) compared to normal plasma cells from healthy donors ( n = 14) (GEO: GSE6477 ) (right). P value was determined by unpaired two-tailed t test. D Quantitative real-time PCR analysis shows LAMP5 expression in normal plasma cells (Healthy, n = 10), malignant plasma cells (Pt MM, n = 20), and human myeloma cell lines (MM cell lines, n = 6). GAPDH served as loading control. P values were determined using one-way ANOVA. E Representative two patients of UMAP plot of CD138 (left) and LAMP5 (right) expression in scRNA-seq data ( GSE161801 ). F Immunofluorescent staining of healthy of myeloma patients bone marrow samples with DAPI and antibodies against LAMP5 and CD138 ( n = 5). Scale bar, 10 μm. G Overall survival (OS) of patient’s myeloma cells with high LAMP5 (High, n = 200) and low LAMP5 (Low, n = 200) expression in MMRF CoMMpass study IA15.

Article Snippet: The CD138 antibody (#AF2780) was purchased from R&D Systems.

Techniques: Expressing, Clinical Proteomics, Two Tailed Test, Real-time Polymerase Chain Reaction, Control, Staining

Journal: Oncogene

Article Title: LAMP5 modulates IRF4 stability and nuclear transport: a critical mechanism in myeloma progression and therapy

doi: 10.1038/s41388-025-03513-x

Figure Lengend Snippet: A Analysis of immunoprecipitates of HA-tagged LAMP5 in IM-9 cells with mass spectrometry. The proteins identified are indicated. B Co-immunoprecipitation of LAMP5 with IRF4 in ARP-1 or IM-9 cells. C Co-immunoprecipitation of LAMP5 with IRF4 in HEK293T cells co-transfected with LAMP5 and IRF4 plasmids. D Pull-down of HA-LAMP5 with IRF4 in HEK293T cells. E Immunofluorescent staining of ARP-1 or IM-9 cells with DAPI and antibodies against LAMP5 and IRF4. Scale bar, 10 μm. F Immunofluorescent staining of myeloma patients bone marrow samples with DAPI and antibodies against CD138, LAMP5 and IRF4 ( n = 5). Scale bar, 10 μm. G Schematic of the truncations including ΔDBD and ΔIAD fragments. H Western blotting showing different truncations of IRF4 (ΔDBD and ΔIAD) in HEK293T cells. I Pull-down of LAMP5 with different truncations of IRF4 (ΔDBD and ΔIAD) in HEK293T cells. J – M The expression of IRF4 or c-MYC in myeloma cells with reduced (sh LAMP5 ) or increased ( LAMP5 ) LAMP5 expression. Nontargeted shRNA (sh Ctrl ) or control vector ( Vec ) served as control. N , O The expression of LDHA or KLF2 in myeloma cells with reduced (sh LAMP5 ) LAMP5 expression. Nontargeted shRNA (sh Ctrl ) served as control. P , Q Immunofluorescent staining of ARP-1 or IM-9 cells (sh Ctrl and sh LAMP5 ) with DAPI and antibody against LAMP5 and IRF4. Scale bar, 10 μm. R Immunofluorescent staining of tumor tissues in Fig. with DAPI and antibodies against LAMP5 and IRF4. Scale bar, 20 μm. S Correlation coefficient of the mRNA levels of LAMP5 and mRNA levels of IRF4 or c-MYC in myeloma patients ( n = 559). The correlations were evaluated using Pearson coefficient. r, correlation coefficient. B – D , H , I , and M are representative of two independent experiments. E , J –L, and N – R are representative of three independent experiments. Data are averages ±SD. P value was determined by Pearson coefficient. J , K , L , N , O P values were determined using one-way ANOVA.

Article Snippet: The CD138 antibody (#AF2780) was purchased from R&D Systems.

Techniques: Mass Spectrometry, Immunoprecipitation, Transfection, Staining, Western Blot, Expressing, shRNA, Control, Plasmid Preparation

Journal: Oncogene

Article Title: LAMP5 modulates IRF4 stability and nuclear transport: a critical mechanism in myeloma progression and therapy

doi: 10.1038/s41388-025-03513-x

Figure Lengend Snippet: A Workflow of high-throughput virtual screening (HTVS) for LAMP5 inhibitors. B Docking scores of the top 4 candidates obtained from HTVS. C Chemical structure and in silico docking of pyrazofurin into the active pocket of human LAMP5 protein. D Proliferation of myeloma cells (ARP-1 or IM-9) treated with or without pyrazofurin (Pyr.) (0.1 or 1 μM) over time. PBS buffer treated group served as control. E Antiproliferative activities of pyrazofurin were evaluated on human myeloma cell lines ARP-1 or IM-9. F Co-immunoprecipitation of LAMP5 with IRF4 in ARP-1 or IM-9 cells treated with or without pyrazofurin (1 μM). G Co-immunoprecipitation of LAMP5 with IRF4 in HEK293T cells co-transfected with HA-LAMP5 and IRF4 plasmids. H IRF4 protein levels in myeloma cells treated with 1 μM pyrazofurin and 2.5 μM MG-132 or 2.5 μM leupeptin for 7 h. 6-week-old male C57BL/6 J mice were intravenously injected vk12598 cells ( n = 9 mice/group), followed by intraperitoneal administration of pyrazofurin (5 mg/kg bodyweight) three times per week for a duration of two weeks, beginning two weeks after cell injection. Shown are the experimental schematic ( I ). J ELISA analysis shown the concentrations of IgG2b in mouse sera. K Immunofluorescent staining of I mentioned spleen tumor cells of vk12598 mice with DAPI and antibodies against CD138 or IRF4. Scale bar, 40 μm. L Photographic images of tumors in NSG mice after implantation of ARP-1 or IM-9 cells ( n = 3 mice/group), and intraperitoneal administration with or without pyrazofurin (5 mg/kg bodyweight). Shown are time course of tumor volume ( M ). N , O Immunofluorescent staining of Ki67 or IRF4 expression in tumor tissues. D and E are representative of three independent experiments. F – H are representative of two independent experiments. Data are averages ±SD. J P value was determined by unpaired two-tailed t test; D , M P values were determined using two-way ANOVA.

Article Snippet: The CD138 antibody (#AF2780) was purchased from R&D Systems.

Techniques: High Throughput Screening Assay, In Silico, Control, Immunoprecipitation, Transfection, Injection, Enzyme-linked Immunosorbent Assay, Staining, Expressing, Two Tailed Test