Journal: Journal of Hematology & Oncology

Article Title: Heterodimerization of T cell engaging bispecific antibodies to enhance specificity against pancreatic ductal adenocarcinoma

doi: 10.1186/s13045-024-01538-5

Figure Lengend Snippet: Tumor-associated antigen expression (normalized MFI, mean fluorescence intensity) in PDAC cell lines

Article Snippet: Either recombinant human EGFR (R&D system: Cat# 9565-ER), HER2 (Sino Biological: Cat# 10,004-H08H), or both were immobilized on CM5 chips.

Techniques: Expressing, Fluorescence

Journal: Journal of Hematology & Oncology

Article Title: Heterodimerization of T cell engaging bispecific antibodies to enhance specificity against pancreatic ductal adenocarcinoma

doi: 10.1186/s13045-024-01538-5

Figure Lengend Snippet: Structure and in vitro activity of PDAC-directed T cell-engaging bispecific antibodies ( A ) IgG-[L]-scFv structure of BsAbs. CH, constant heavy chain; CL, constant light chain; scFv, single chain variable fragment; VH, variable heavy chain; VL, variabl light chain. ( B ) Flow cytometric analysis displaying fluorescent intensities of IgG-[L]-scFv BsAbs bound to human PDAC cells. IgG-[L]-scFv BsAbs evaluated were specific for human EGFR, HER2, MSLN, c-MET, and B7-H3. ( C ) Antibody-dependent T cell-mediated cytotoxicity (ADTC) as a function of increasing doses of BsAbs against PDAC cell lines. Ratio of effector T cells to target PDAC cells (E: T ratio) was set to 10:1

Article Snippet: Either recombinant human EGFR (R&D system: Cat# 9565-ER), HER2 (Sino Biological: Cat# 10,004-H08H), or both were immobilized on CM5 chips.

Techniques: In Vitro, Activity Assay

Journal: Journal of Hematology & Oncology

Article Title: Heterodimerization of T cell engaging bispecific antibodies to enhance specificity against pancreatic ductal adenocarcinoma

doi: 10.1186/s13045-024-01538-5

Figure Lengend Snippet: In vitro sensitivities (EC50, pM) to target antigen-specific bispecific antibodies in PDAC cell lines

Article Snippet: Either recombinant human EGFR (R&D system: Cat# 9565-ER), HER2 (Sino Biological: Cat# 10,004-H08H), or both were immobilized on CM5 chips.

Techniques: In Vitro

Journal: Journal of Hematology & Oncology

Article Title: Heterodimerization of T cell engaging bispecific antibodies to enhance specificity against pancreatic ductal adenocarcinoma

doi: 10.1186/s13045-024-01538-5

Figure Lengend Snippet: EGFR and HER2 T-BsAb heterodimerization and in vitro kinetics ( A ) Schematic of BsAb heterodimerization by controlled Fab Arm Exchange. Parental BsAbs bore 2 Fabs specific to EGFR or HER2 and 2 scFvs specific for CD3 (‘2 + 2’). Reduction of inter-H-chain disulfide bonds and subsequent heterodimerization (driven by F405L and K409R amino acid substitutions) yielded EGFRxHER2 BsAbs bearing 1 EGFR Fab, 1 HER2 Fab, and 2 CD3 scFv (‘1 + 1 + 2’). ( B ) SPR analysis of T-BsAbs binding to EGFR (left panel), HER2 (middle panel), and EGFR x HER2 (right panel) proteins. Representative normalized sensorgrams at 20nM were shown. ( C ) IHC staining of two cell-derived PDAC xenograft tumors SW1990 (top) and BxPC-3 (bottom). OCT-embedded tumor sections were stained with EGFRxEGFR, HER2xHER2, or EGFRxHER2 BsAbs. Control slides were either unstained (SW1990) or incubated with a control antibody (BxPC-3). ( D ) Antibody-dependent T cell-mediated cytotoxicity assays (ADTC) against SW1990 (left) and BxPC-3 (right). Cytotoxicity measured by Chromium-51 release. Ratio of effector T cells to target PDAC cells (E: T ratio) was set to 10:1. EC50s (SW1990; BxPC-3): EGFRxEGFR: 165.8fM; 31.5fM, HER2xHER2: 18.5pM; 7.14pM, EGFRxHER2: 699.6fM; 128.8fM. CD33xCD33 BsAb was included as a negative control

Article Snippet: Either recombinant human EGFR (R&D system: Cat# 9565-ER), HER2 (Sino Biological: Cat# 10,004-H08H), or both were immobilized on CM5 chips.

Techniques: In Vitro, Binding Assay, Immunohistochemistry, Derivative Assay, Staining, Control, Incubation, Negative Control

Journal: Journal of Hematology & Oncology

Article Title: Heterodimerization of T cell engaging bispecific antibodies to enhance specificity against pancreatic ductal adenocarcinoma

doi: 10.1186/s13045-024-01538-5

Figure Lengend Snippet: Avidities of EGFR and HER2 T-BsAbs

Article Snippet: Either recombinant human EGFR (R&D system: Cat# 9565-ER), HER2 (Sino Biological: Cat# 10,004-H08H), or both were immobilized on CM5 chips.

Techniques: Protein Binding

Journal: Journal of Hematology & Oncology

Article Title: Heterodimerization of T cell engaging bispecific antibodies to enhance specificity against pancreatic ductal adenocarcinoma

doi: 10.1186/s13045-024-01538-5

Figure Lengend Snippet: Heterodimeric EGFR and HER2 T-BsAbs impede PDAC growth ( A ) In vivo antitumor effect of BsAbs in the presence of human T cells against SW1990 cell line xenografts; 3 × 10 6 cells of SW1990 were subcutaneously implanted into mice. T-BsAbs were administered twice per week and 2 × 10 7 of luciferase-transduced T cells (Luc(+) T cells) were administered once per week for 2 weeks to treat the tumors. ( B ) In vivo anti-tumor effects of 10 µg EGFR and HER2 T-BsAbs. ( C ) Relative body weight of mice during treatment and ( D ) overall survival were plotted. ( E ) Bioluminescence imaging (BLI) of Luc(+) T cells. Quantitation of bioluminescence intensity in the lesions of tumors. Representative images (right) were taken on day 7

Article Snippet: Either recombinant human EGFR (R&D system: Cat# 9565-ER), HER2 (Sino Biological: Cat# 10,004-H08H), or both were immobilized on CM5 chips.

Techniques: In Vivo, Luciferase, Imaging, Quantitation Assay

Journal: Journal of Hematology & Oncology

Article Title: Heterodimerization of T cell engaging bispecific antibodies to enhance specificity against pancreatic ductal adenocarcinoma

doi: 10.1186/s13045-024-01538-5

Figure Lengend Snippet: T-BsAbs drive T cell infiltration, activation, and function in PDAC ( A ) Treatment of SW1990 tumor-bearing mice with EGFR and HER2 T-BsAbs; 3 × 10 6 cells of SW1990 were subcutaneously implanted into mice. Once tumor reached ∼500mm 3 , mice received a single infusion of 2 × 10 7 T cells and were treated with 10µg T-BsAb, administered twice per week. Tumors were harvested for analysis on day 10 after initiation of treatment. Mice that exhibited 50% or more reduction in tumor volume by day 10 were excluded from this and subsequent analyses. ( B ) Flow cytometric analysis of the in vivo effect of T-BsAb treatments on T cell infiltration into harvested SW1990 PDAC tumors. Infiltrating T cells were identified by detection of human CD45 + and human CD4 + or CD8 + . ( C ) Frequency of T cell activation indicated by detection of IFN-γ + TNF-α + T cells and expression of CD69 among CD4 + or CD8 + T cells. ( D ) Frequency of CD11b + intratumoral myeloid cells and prevalence of PDL1 expression

Article Snippet: Either recombinant human EGFR (R&D system: Cat# 9565-ER), HER2 (Sino Biological: Cat# 10,004-H08H), or both were immobilized on CM5 chips.

Techniques: Activation Assay, In Vivo, Expressing

Journal: Journal of Hematology & Oncology

Article Title: Heterodimerization of T cell engaging bispecific antibodies to enhance specificity against pancreatic ductal adenocarcinoma

doi: 10.1186/s13045-024-01538-5

Figure Lengend Snippet: ADTC sensitivities (EC50, pM) of SW1990 lines to EGFR and HER2 T-BsAbs

Article Snippet: Either recombinant human EGFR (R&D system: Cat# 9565-ER), HER2 (Sino Biological: Cat# 10,004-H08H), or both were immobilized on CM5 chips.

Techniques:

Journal: Journal of Hematology & Oncology

Article Title: Heterodimerization of T cell engaging bispecific antibodies to enhance specificity against pancreatic ductal adenocarcinoma

doi: 10.1186/s13045-024-01538-5

Figure Lengend Snippet: FACS binding (EC50, pM) of EGFR and HER2 T-BsAbs to SW1990 lines

Article Snippet: Either recombinant human EGFR (R&D system: Cat# 9565-ER), HER2 (Sino Biological: Cat# 10,004-H08H), or both were immobilized on CM5 chips.

Techniques: Binding Assay

Journal: Journal of Hematology & Oncology

Article Title: Heterodimerization of T cell engaging bispecific antibodies to enhance specificity against pancreatic ductal adenocarcinoma

doi: 10.1186/s13045-024-01538-5

Figure Lengend Snippet: EGFRxHER2 T-BsAbs efficacy is lost in EGFR and HER2-single-positive PDAC ( A ) Mean fluorescent intensities determined by flow cytometry and ( B ) antibody-dependent T cell-mediated cytotoxicity (ADTC) against SW1990-EGFR-KO (left) and SW1990-HER2-KO (right) cell lines. Ratio of effector T cells to target PDAC cells (E: T ratio) was set to 10:1. ( C ) Tumor growth of subcutaneous SW1990 EGFR-KO (left) and HER2-KO (right) tumors. One infusion of 2 × 10 7 T cells was administered. Two doses of T-BsAbs (10 µg each) were given by i.v. injection

Article Snippet: Either recombinant human EGFR (R&D system: Cat# 9565-ER), HER2 (Sino Biological: Cat# 10,004-H08H), or both were immobilized on CM5 chips.

Techniques: Flow Cytometry, Injection

Journal: bioRxiv

Article Title: Zinc Alpha-2-Glycoprotein (ZAG/AZGP1) secreted by triple-negative breast cancer promotes tumor microenvironment fibrosis

doi: 10.1101/2024.03.04.583349

Figure Lengend Snippet: (A) Inhibition of 3T3-L1 adipogenesis is rescued with heat inactivation of the secretome of MDA-MB-468 and MDA-MB-231 cells. (B) The anti-adipogenic factor of the MDA-MB-468 and MDA-MB-231 secretomes (total) is retained in the supernatant (top) fraction following centrifugation using a spin column with a molecular weight cut-off of 100,000 Daltons. (C) Candidate anti-adipogenic factors identified in the secretome of MDA-MB-468 and MDA-MB-231 cells. (D) The secretome of MDA-MB-468 cells depleted of ZAG does not inhibit 3T3-L1 adipogenesis. (E) Supplementing 3T3-L1 cells with recombinant ZAG during the first two days of adipogenesis is sufficient to inhibit adipogenesis. (A-E) All data are mean ± SD. Data points show independent biological replicates. p-values calculated using one-way ANOVA followed by (A-B) Tukey’s multiple comparison test or (D) Dunnett’s multiple comparison test, or (E) two-way ANOVA followed by Šidák’s multiple comparison test. (ns is non-significant; *<0.05, **<0.01, and ****<0.0001). See also Figure S2.

Article Snippet: Where indicated, differentiation media was supplemented with recombinant ZAG (R&D Systems #4764-ZA) or TSP-1 (R&D Systems #3074-TH).

Techniques: Inhibition, Centrifugation, Molecular Weight, Recombinant, Comparison

Journal: Cells

Article Title: GDF3 Protects Mice against Sepsis-Induced Cardiac Dysfunction and Mortality by Suppression of Macrophage Pro-Inflammatory Phenotype

doi: 10.3390/cells9010120

Figure Lengend Snippet: Higher serum levels of GDF3 are related to poor outcomes of septic patients. ( A ) Scatter plots of serum GDF3 levels on admission of healthy donors and septic patients. ( B ) AUC discriminating sepsis from healthy controls. ( C , D ) Correlation between serum GDF3 levels and ( C ) APACHE II score and ( D ) SOFA score at admission of ICU. ( E ) Serum GDF3 levels in different groups sorted by survivor and non-survivor. ( F ) AUROC predicting 28-day mortality: GDF3 (AUC 0.770), Lac (AUC 0.767), CRP (AUC 0.632), PCT (AUC 0.677), SAA (AUC 0.724). (*, p < 0.05 vs. healthy donors; #, p < 0.05 vs. survivor).

Article Snippet: Recombinant mouse GDF3 protein (R&D Systems, Minneapolis, MN, USA) was added with BSA as a carrier protein to enhance protein stability and increase shelf-life.

Techniques:

Journal: Cells

Article Title: GDF3 Protects Mice against Sepsis-Induced Cardiac Dysfunction and Mortality by Suppression of Macrophage Pro-Inflammatory Phenotype

doi: 10.3390/cells9010120

Figure Lengend Snippet: The dynamic expression of GDF3 in mice and macrophages treated with endotoxin. ( A ) mRNA levels of GDF3 in different tissues of mice, n = 4. GAPDH gene expression was used as the internal control. ( B ) Serum GDF3 levels are detected at indicated time points in mice after LPS (10mg/kg) injection (*, p < 0.05; n = 4). mRNA levels of GDF3 were determined in ( C ) whole blood and ( D ) spleens of LPS-injected mice (*, p < 0.05; n = 6). ( E ) LPS-dose response and ( F ) time course of GDF3 expression in BMDMs, where indicated, ( E ) total RNAs were collected at 24 h after LPS exposure (*, p < 0.05) and ( F ) BMDMs were pre-treated with LPS (10 ng/mL) (*, p < 0.05; n = 4). Similar results were obtained in other two independent experiments.

Article Snippet: Recombinant mouse GDF3 protein (R&D Systems, Minneapolis, MN, USA) was added with BSA as a carrier protein to enhance protein stability and increase shelf-life.

Techniques: Expressing, Gene Expression, Control, Injection

Journal: Cells

Article Title: GDF3 Protects Mice against Sepsis-Induced Cardiac Dysfunction and Mortality by Suppression of Macrophage Pro-Inflammatory Phenotype

doi: 10.3390/cells9010120

Figure Lengend Snippet: Recombinant GDF3 protein suppresses pro-inflammatory cytokine production in LPS-treated macrophages. ( A – D ) BMDMs treated with the indicated doses of recombinant GDF3 (rGDF3) and ( E – H ) RAW264.7 macrophages treated with rGDF3 (50 ng/mL) for 2 h, followed by addition of LPS (10 ng/mL) for 24 h. Culture supernatants were measured for ( A , E ) TNF-α, ( B , F ) IL-6, ( C , G ) IL-1β and ( D , H ) MCP-1 by ELISA (*, p < 0.05; n = 7–9 wells). Similar results were obtained in another separated experiments.

Article Snippet: Recombinant mouse GDF3 protein (R&D Systems, Minneapolis, MN, USA) was added with BSA as a carrier protein to enhance protein stability and increase shelf-life.

Techniques: Recombinant, Enzyme-linked Immunosorbent Assay

Journal: Cells

Article Title: GDF3 Protects Mice against Sepsis-Induced Cardiac Dysfunction and Mortality by Suppression of Macrophage Pro-Inflammatory Phenotype

doi: 10.3390/cells9010120

Figure Lengend Snippet: Recombinant GDF3 protein reduces macrophage M1 and increases M2 polarization in LPS-treated BMDMs. BMDMs were treated with rGDF3 (50 ng/mL) for 2 h, then added LPS (10 ng/mL) for 6 h. mRNA levels of ( A ) iNOS and ( B ) Arg-1 were evaluated by RT-PCR. The mRNA levels were normalized to 18S mRNA levels and expressed as fold versus BSA group (*, p < 0.05 vs. Ctl; #, p < 0.05 vs. LPS; n = 6). ( C , D ) BMDMs were treated with rGDF3 (50 ng/mL) for 2 h, followed by LPS (10 ng/mL) exposure for 12 h. These BMDMs were stained with antibodies to ( C ) CD38 and ( D ) CD206 for flow cytometry analysis. Quantification results for ( E ) CD38 positive macrophages and ( F ) CD206 positive macrophages. (*, p < 0.05 vs. BSA; #, p < 0.05 vs. LPS; n = 3). Similar results were obtained in other two separated experiments.

Article Snippet: Recombinant mouse GDF3 protein (R&D Systems, Minneapolis, MN, USA) was added with BSA as a carrier protein to enhance protein stability and increase shelf-life.

Techniques: Recombinant, Reverse Transcription Polymerase Chain Reaction, Staining, Flow Cytometry

Journal: Cells

Article Title: GDF3 Protects Mice against Sepsis-Induced Cardiac Dysfunction and Mortality by Suppression of Macrophage Pro-Inflammatory Phenotype

doi: 10.3390/cells9010120

Figure Lengend Snippet: Recombinant GDF3 protein attenuates inflammatory response and mortality in endotoxin-induced septic mice. WT mice injected with rGDF3 protein (I.P., 10μg/kg BW) 12 h prior to LPS injection (I.P., 10mg/kg BW), 12 h later serum was collected for measuring ( A ) TNF-α, ( B ) IL-6, and ( C ) MCP-1 levels (*, p < 0.05 vs. Ctl; #, p < 0.05 vs. LPS + B; n = 5). ( D ) Kaplan-Meier survival curves were generated for LPS-mice treated with or without rGDF3 (*, p < 0.05; n = 12).

Article Snippet: Recombinant mouse GDF3 protein (R&D Systems, Minneapolis, MN, USA) was added with BSA as a carrier protein to enhance protein stability and increase shelf-life.

Techniques: Recombinant, Injection, Generated

Journal: Cells

Article Title: GDF3 Protects Mice against Sepsis-Induced Cardiac Dysfunction and Mortality by Suppression of Macrophage Pro-Inflammatory Phenotype

doi: 10.3390/cells9010120

Figure Lengend Snippet: Administration of rGDF3 to LPS-mice improves cardiac function and promotes cardiac macrophage M2 phenotype. WT mice received recombinant GDF3 protein (I.P., 10μg/kg BW) 12 h prior to LPS injection (I.P., 10mg/kg BW), 12 h later cardiac function was measured by echocardiograph. ( A ) Representative M-mode echocardiography recordings for three groups, and ( B ) left ventricular ejection fraction (EF %), ( C ) fractional shortening (FS %) and ( D ) left ventricular internal dimension at end-systolic (LVIDs) were calculated (*, p < 0.05 vs. Ctl; #, p < 0.05 vs. LPS+B; n = 5). Cardiac cells were stained with antibodies to ( E ) MHC-II, ( G ) CD206, ( I ) F4/80 plus CD11b, ( K ) Ly6C for flow cytometry analysis and their respective quantification results are shown in ( F , H , J , L ). (*, p < 0.05; n = 4). Similar results were obtained in other two independent experiments.

Article Snippet: Recombinant mouse GDF3 protein (R&D Systems, Minneapolis, MN, USA) was added with BSA as a carrier protein to enhance protein stability and increase shelf-life.

Techniques: Recombinant, Injection, Staining, Flow Cytometry

Journal: Cells

Article Title: GDF3 Protects Mice against Sepsis-Induced Cardiac Dysfunction and Mortality by Suppression of Macrophage Pro-Inflammatory Phenotype

doi: 10.3390/cells9010120

Figure Lengend Snippet: GDF3-mediated macrophage M2 phenotype is associated with the activation of Samd2/3 and inhibition of NLRP3 inflammasome. ( A ) Representative immuno-blots and quantification results for ( B ) p-Samd2, ( C ) p-Smad3 and ( D ) NLRP3. GAPDH was used as a loading control for total protein (*, p < 0.05 vs. Ctl cells, #, p < 0.05 vs. LPS treated cells; n = 3). ( E – G ) RAW264.7 macrophages were pre-treated with SB431542 or DMSO (10μM) for 0.5 h, then incubated with rGDF3 (50 ng/mL) or BSA for 1 h, followed by stimulation with LPS (10 ng/mL) for 12 h. Supernatants were collected to measure ( E ) TNF-α, ( F ) IL-6, and ( G ) IL-1β levels (*, p < 0.05 vs. Ctl cells; #, p < 0.05 vs. LPS-treated cells; n = 4). The mRNA levels of ( H ) iNOS and ( I ) Arg-1 in these macrophages treated as above were measured by RT-PCR. The fold changes were normalized to 18S (*, p < 0.05 vs. LPS-cells; n = 4). Data shown were representative of three independent experiments.

Article Snippet: Recombinant mouse GDF3 protein (R&D Systems, Minneapolis, MN, USA) was added with BSA as a carrier protein to enhance protein stability and increase shelf-life.

Techniques: Activation Assay, Inhibition, Western Blot, Control, Incubation, Reverse Transcription Polymerase Chain Reaction

Journal: Cells

Article Title: GDF3 Protects Mice against Sepsis-Induced Cardiac Dysfunction and Mortality by Suppression of Macrophage Pro-Inflammatory Phenotype

doi: 10.3390/cells9010120

Figure Lengend Snippet: Scheme depicting GDF3-drived protection against endotoxin-induced inflammation, cardiac dysfunction and mortality. Exogenous GDF3 binds to ALK4/5/7 where activates Smad2/3 and inhibits NLRP3 inflammasome, consequently suppresses macrophage pro-inflammatory phenotype (M1), leading to reduced inflammation, cardiac dysfunction, and mortality in endotoxin-induced septic mice.

Article Snippet: Recombinant mouse GDF3 protein (R&D Systems, Minneapolis, MN, USA) was added with BSA as a carrier protein to enhance protein stability and increase shelf-life.

Techniques:

Journal: PLoS ONE

Article Title: Networked T Cell Death following Macrophage Infection by Mycobacterium tuberculosis

doi: 10.1371/journal.pone.0038488

Figure Lengend Snippet: (A) Addition of the pan-caspase inhibitor Z-VAD-FMK mitigated cell death induced by low MOI and high MOI-infected macrophage supernatants (shaded bars, infection + and ++ respectively), as well as background cell death observed in the uninfected control supernatants (closed bars, infection -). Cell death in infected supernatants was not mitigated by coincubation with blocking antibodies to TNF-α (B) or Fas (C), and in each case was significantly higher than in uninfected control supernatants (closed bars). Cell death induced by positive controls (D) (1 µM Staurosporine, 5 ng/ml recombinant human TNF-α +0.2 µg/ml cycloheximide and 10 ng/ml recombinant human Fas ligand) was in each case significantly reduced by coincubation with the corresponding inhibitor (open bars). Data shown in each panel are from one representative experiment, showing mean percentage cell death ± SEM from triplicate incubations. ** = p<0.01, *** = p<0.001, Student’s t test relative to uninfected control (A–C), or positive control relative to inhibitor (D).

Article Snippet: In positive control experiments for Fas ligand-mediated cell killing, cells were incubated in the presence of 10 ng/ml recombinant human Fas ligand (R&D systems), crosslinked with 10 μg/ml anti-6× Histidine (R&D systems).

Techniques: Infection, Control, Blocking Assay, Recombinant, Positive Control

Journal: Mediators of Inflammation

Article Title: Cytomegalovirus Infection Impairs Immunosuppressive and Antimicrobial Effector Functions of Human Multipotent Mesenchymal Stromal Cells

doi: 10.1155/2014/898630

Figure Lengend Snippet: CMV blocks IDO activity and IDO-mediated antimicrobial effects in human MSC. (a) PBL, stimulated with OKT3, were cocultured with MSC in the presence or absence of CMV. As controls UV-inactivated CMV (uvCMV), the IDO-specific inhibitor 1-L-methyl-tryptophan (1-MT; 1.5 mM), or a neutralising anti-IFN- γ antibody ( α IFN- γ ; 10 ng/mL) was used. After three days IDO activity was determined and is presented as mean kynurenine content ± SEM of three independent experiments, each done in triplicate. (b) PBL (1 × 10 5 /well), stimulated with CD3-directed mAB OKT3, were cocultured with MSC (3 × 10 4 /well) in the absence or presence of CMV (MOI 5). After three days cultures were infected with S. aureus (10–100 cfu/well) and bacterial growth was determined photometrically. As a control, cultures were supplemented with L-tryptophan (Trp; 0.6 mM) at the time point of bacterial infection. Data are given as mean OD (620 nm) ± SEM of three experiments, each done in triplicate. Significant differences ( P < 0.05) as compared to the positive control are marked by asterisks.

Article Snippet: Recombinant human IFN- γ was purchased from R&D Systems (Wiesbaden, Germany).

Techniques: Activity Assay, Infection, Control, Positive Control

Journal: Mediators of Inflammation

Article Title: Cytomegalovirus Infection Impairs Immunosuppressive and Antimicrobial Effector Functions of Human Multipotent Mesenchymal Stromal Cells

doi: 10.1155/2014/898630

Figure Lengend Snippet: CMV inhibits IDO induction by recombinant IFN- γ . (a) MSC (2 × 10 4 /well) which were infected with various amounts of CMV (MOI 0.1–10) were stimulated with IFN- γ (300 U/mL). After three days IDO activity was determined and is presented as mean kynurenine content ± SEM of three independent experiments, each done in triplicate. (b) MSC (1.5 × 10 6 /flask) were stimulated with IFN- γ (600 U/mL) in the absence or presence of CMV (MOI 5). Cells were harvested after 24 h and IDO protein was detected in Western blot analysis. β -Actin was utilized as a protein loading control, while the viral pp72 protein served as an infection control.

Article Snippet: Recombinant human IFN- γ was purchased from R&D Systems (Wiesbaden, Germany).

Techniques: Recombinant, Infection, Activity Assay, Western Blot, Control