Journal:

Article Title: CTLA-4 regulates allergen response by modulating GATA-3 protein level per cell

doi: 10.1111/j.1365-2567.2007.02537.x

Figure Lengend Snippet: Effects of in vitro CTLA-4 blockade on GATA-3 protein level/cell and frequency of IL-4-producing cells. Naive CD4+ T cells purified from the spleens of normal mice were induced to differentiate with anti-CD3 mAb, a recombinant form of CD80-Fc, IL-4 and anti-IFN-γ mAb for 3 days. Anti-CTLA-4 mAb was added in soluble form at increasing concentrations (0·1–10 μg/ml). (a) Mean fluorescence intensity for GATA-3 staining, (b) percentage of IL-4+ cells.

Article Snippet: 18 CD4 cell culture For in vitro experiments, splenic naive CD4 cells were purified from non-immunized BALB/c mice by immuno-magnetic cell sorting as previously described 19 and cultured in 24-well plates precoated with anti-CD3ε mAb (clone 145-2C11; 10 μg/ml) and/or recombinant mouse B7-1(CD80)/Fc chimeric protein (R & D Systems, cat. 740-B1; 0·33 μg/ml).

Techniques: In Vitro, Purification, Recombinant, Fluorescence, Staining

Journal: Current protocols

Article Title: Redefining Cell Culture Using a 3D Flipwell Co-culture System: A Mimetic for Gut Architecture and Dynamics In Vitro.

doi: 10.1002/cpz1.70107

Figure Lengend Snippet: Figure 15 THP-1 dramatic polarization toward an M1-like phenotype with Sepiapterin treatment in the 3D Flipwell coculture system.(A) Confocal images of THP-1 cells cocultured with Caco-2:HT- 29 in the 3D Flipwells and stained for pan macrophage marker CD68 (red) and CD80 (green). Pan macrophage CD68 marker is present in both the SEP treated sample and the untreated control. CD80 marker, typical of the M1 polarization phenotype, is more prominently present in the SEP treated sample with long pseudopodia extending outward typical of the M1 phenotype. (B) Con- focal images of THP-1 cells cocultured with Caco-2:HT-29 in the 3D Flipwell and stained for pan macrophage marker CD68 and CD163 to validate phenotypic polarization toward M1 phenotype and away from M2 phenotype. Scale bar = 25 μm.

Article Snippet: See the T erm s and C onditions (https://onlinelibrary.w iley.com s-and-conditions) on W iley O nline L ibrary for rules of use; O A articles are governed by the applicable C reative C om m ons L icense Anti-human MUC2 (996/1) (Thermo Fisher, cat. no. MA5-12345) Anti-human CK20 (Life Technologies, cat. no. PA5-82875) Anti-human CD80 (Cell Signaling Technologies, E3Q9V, cat. no. 15416) Anti-human CD163 (Abcam, cat. no. 156769) or anti-human CD68 (Novus Biologicals, cat. no. NB100-683) Secondary antibodies: Alexa Fluor 488 goat anti-rabbit IgG (Thermo Fisher, cat. no. A11008) for CD80 and CK20 Alexa Fluor 594 goat anti-mouse IgG (Thermo Fisher, cat. no. A11005) for MUC2 and CD68 or CD163 DAPI, VectaShield mounting medium (Vector Laboratories, cat. no. H-1000) 50-ml conical tubes (SpectraTube, Alkali Scientific cat. no. C2750) Plastic wrap Lab tape or shipping tape Tweezers 12-well tissue culture plate (Corning Costar, cat. no. 3513) 6-well dish Aluminum foil Scalpel Round cover slips, #1, 1 oz (Corning Float Glass, cat. no. 50143822, or Fisherbrand cat. no. 12-546-2 25CIR-2) Laboratory marker (VWR, cat. no 95042-566) Attofluor cell chamber coverslip holder (Thermo Fisher Scientific, cat. no. A7816) Confocal microscope and software [e.g., Leica Microsystems TCS SP5, multi-photon laser scanning, and Suite Advanced Fluorescence (LAS AF) software] Deep Petri dish bottoms, 100 × 25–mm deep polystyrene stackable Petri dish, sterile (USA Scientific, cat. no. 8609-0625) 10-, 200-, and 1000-μl pipet tips (Alkali Scientific PurePoint or equivalent) Beckman Coulter Allegra 6 centrifuge with a swing bucket rotor for 50-ml conical tubes Optional: ImmunoPen (Fisher Scientific, cat. no. 4021761EA) Wash, permeabilization, disassembly, and blocking steps 1.

Techniques: Staining, Marker, Control

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Co-targeting CD47 and VEGF elicited potent anti-tumor effects in gastric cancer

doi: 10.1007/s00262-024-03667-9

Figure Lengend Snippet: Targeting CD47 significantly inhibited the growth of gastric cancer in Hu-PDX models. A In the Hu-PDX1 model, tumor volume was measured twice a week and presented as mean ± SD. After treatment with SIRPα-Fc for 4 weeks, tumor weight was presented. ( n = 6 for control group and n = 8 for SIRPα-Fc group, * P < 0.05, ** P < 0.01) B Each line represented the tumor volume from an independent mouse in the Hu-PDX1 model. C In the Hu-PDX2 model, tumor volume was measured twice a week and presented as mean ± SD. After treatment with SIRPα-Fc for 4 weeks, tumor weight was presented. ( n = 6 per group, * P < 0.05, ** P < 0.01). D Each line represented the tumor volume from an independent mouse in the Hu-PDX2 model. E Representative photographs of immunohistochemical staining for CD80, CD163, and CD8 of tumor tissue sections and the number of CD80 + , CD163 + , and CD8 + cells in each group were normalized to the control group. The value of control was set to 1.0. ( * P < 0.05, ** P < 0.01)

Article Snippet: Anti-CD80 monoclonal antibody (66,406-1-Ig, Proteintech), anti-CD163 monoclonal antibody (GB13340, Servicebio), anti-CD8 monoclonal antibody (GB12068, Servicebio), anti-CD31 monoclonal antibody (GB113151, Servicebio), VEGFA Monoclonal antibody (19,003-1-AP, Proteintech), anti-CD47 monoclonal antibody (ab218810, Abcam), carboxyfluorescein diacetate succinimidyl ester (CFDA SE) (C0051, Beyotime), PerCP anti-CD68 (333,813, BioLegend), PE anti-CD11b (101,208, BioLegend), granulocyte–macrophage colony-stimulating factor (GM-CSF) (C003, novoprotein), FITC-labeled anti-CD47 (CC2C6, BioLegend), Human Lymphocyte separation medium (7,111,011, DAKEWE).

Techniques: Control, Immunohistochemical staining, Staining

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Co-targeting CD47 and VEGF elicited potent anti-tumor effects in gastric cancer

doi: 10.1007/s00262-024-03667-9

Figure Lengend Snippet: CD47 blockade combined with antiangiogenetic therapy elicited enhanced anti-tumor effect in Hu-PDX models of gastric cancer. A and B In the Hu-PDX1 model, tumor-bearing mice were treated with SIRPα-Fc and/or VEGFR1-Fc for 4 weeks, tumor volume and tumor weight were presented as mean ± SD. Each line represented the value of the tumor volume of a single mouse. C and D In the Hu-PDX2 model, tumor volume and tumor weight were presented after the same treatment in the Hu-PDX1 model. Each line represented the value of the tumor volume of a single mouse. ( n = 6 per group, * P < 0.05, ** P < 0.01). E Representative photographs of immunohistochemical staining for CD80, CD163, CD8, and CD31 of tumor tissue sections and the number of CD80 + , CD163 + , and CD8 + cells and the relative vessel density in each group were normalized to the control group. The value of control was set to 1.0. (* P < 0.05, ** P < 0.01)

Article Snippet: Anti-CD80 monoclonal antibody (66,406-1-Ig, Proteintech), anti-CD163 monoclonal antibody (GB13340, Servicebio), anti-CD8 monoclonal antibody (GB12068, Servicebio), anti-CD31 monoclonal antibody (GB113151, Servicebio), VEGFA Monoclonal antibody (19,003-1-AP, Proteintech), anti-CD47 monoclonal antibody (ab218810, Abcam), carboxyfluorescein diacetate succinimidyl ester (CFDA SE) (C0051, Beyotime), PerCP anti-CD68 (333,813, BioLegend), PE anti-CD11b (101,208, BioLegend), granulocyte–macrophage colony-stimulating factor (GM-CSF) (C003, novoprotein), FITC-labeled anti-CD47 (CC2C6, BioLegend), Human Lymphocyte separation medium (7,111,011, DAKEWE).

Techniques: Immunohistochemical staining, Staining, Control

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Co-targeting CD47 and VEGF elicited potent anti-tumor effects in gastric cancer

doi: 10.1007/s00262-024-03667-9

Figure Lengend Snippet: Bispecific fusion protein SIRPα-VEGFR1 elicited synergetic antitumor effect and prevented gastric cancer recurrence. A and B In the Hu-PDX1 model, tumor volume and tumor weight were measured and the data was presented as mean ± SD after treatment with SIRPα-Fc plus VEGFR1-Fc, and SIRPα-VEGFR1 for 4 weeks. Each line represented the value of the tumor volume of a single mouse. C and D In the Hu-PDX2 model, tumor volume and tumor weight were measured after the same treatment in the Hu-PDX1 model. E The number of CD80 + , CD163 + , and CD8 + cells and the relative vessel density in each group were normalized to the control group. The value of control was set to 1.0. (* P < 0.05, ** P < 0.01). F In the humanized tumor recurrence model, mice were treated with control, SIRPα-Fc + VEGFR1-Fc, SIRPα-VEGFR1 for 2 weeks, and tumor volume was measured. Each line represented the value of the tumor volume of a single mouse. G Survival curves for different treatment groups. (n = 6 per group, * P < 0.05, ** P < 0.01)

Article Snippet: Anti-CD80 monoclonal antibody (66,406-1-Ig, Proteintech), anti-CD163 monoclonal antibody (GB13340, Servicebio), anti-CD8 monoclonal antibody (GB12068, Servicebio), anti-CD31 monoclonal antibody (GB113151, Servicebio), VEGFA Monoclonal antibody (19,003-1-AP, Proteintech), anti-CD47 monoclonal antibody (ab218810, Abcam), carboxyfluorescein diacetate succinimidyl ester (CFDA SE) (C0051, Beyotime), PerCP anti-CD68 (333,813, BioLegend), PE anti-CD11b (101,208, BioLegend), granulocyte–macrophage colony-stimulating factor (GM-CSF) (C003, novoprotein), FITC-labeled anti-CD47 (CC2C6, BioLegend), Human Lymphocyte separation medium (7,111,011, DAKEWE).

Techniques: Control

Journal: Arthritis Research & Therapy

Article Title: Interferon-induced protein IFIT4 is associated with systemic lupus erythematosus and promotes differentiation of monocytes into dendritic cell-like cells

doi: 10.1186/ar2475

Figure Lengend Snippet: Comparison of the phenotypic profiles of DCLCs primed with or without IFIT4 over-expression. (a) Surface antigens of normal THP-1 cells were examined by flow cytometry. (b) To analyze the effect of IFIT4 on phenotypic changes of dendritic cell-like cells (DCLCs) during the process of differentiation, THP-1 cells were transfected with pEGFP-IFIT4 or pEGFP-C1; 36 hours later, cells were stimulated with granulocyte-macrophage colony-stimulating factor (GM-CSF; 50 ng/ml) and IL-4 (20 ng/ml) for 90 hours to generate DCLCs. These DCLCs primed with pEGFP-IFIT4 or pEGFP-C1 transfection were incubated with fluorochrome-conjugated monoclonal antibodies (mAbs) and the antigens of CD40, CD80, CD86, CD83, HLA-DR, CD14 and CD1a on the surface of those DCLCs were analyzed by flow cytometry. Appropriate fluorochrome or isotype control mAbs of each antibody species were used as negative controls. Shaded histograms represent isotype control antibodies. The thick line represents DCLCs primed with pEGFP-IFIT4 transfection, whereas the slender lines represent DCLCs primed with pEGFP-C1 transfection. All experiments were performed three times and a set of representative histograms was presented. IFIT4, interferon-induced protein with tetratricopeptide repeats 4.

Article Snippet: FITC anti-human CD40 (Catalogue no: FAB617F) and PE anti-human CD80 (Catalogue no: FAB140P) were from R&D Systems, Minneapolis, MN, USA.

Techniques: Comparison, Over Expression, Flow Cytometry, Transfection, Incubation, Bioprocessing, Control

Journal: Frontiers in Immunology

Article Title: Impact of alcohol consumption on outcomes and potential of immune biomarkers for postoperative complications in trauma patients

doi: 10.3389/fimmu.2025.1492288

Figure Lengend Snippet: Comparison of patient demographics, comorbidities, and postoperative complications between alcohol-negative and alcohol-positive groups included in the ELISA analysis.

Article Snippet: ELISA kits for CD28 (#DY342-05), B7-1 (#DY140) (R&D Systems, Minneapolis, MN, USA), Interleukin 13 (IL-13; #900-K23), Eotaxin-3 (#900-K167), and Tissue Inhibitor of Metalloproteinase 1 (TIMP-1; #900-K438) (PeproTech, Hamburg, Germany) were used according to the manufacturer’s protocols.

Techniques: Comparison, Enzyme-linked Immunosorbent Assay, Control, Infection

Journal: Frontiers in Immunology

Article Title: Impact of alcohol consumption on outcomes and potential of immune biomarkers for postoperative complications in trauma patients

doi: 10.3389/fimmu.2025.1492288

Figure Lengend Snippet: ELISA quantification of key inflammatory and T/B cell activation markers in patient serum samples. Patients were categorized as negative for alcohol risk (Neg.) or positive for alcohol risk (Pos.). Each group is subdivided into a control group (Ctr.) and a complication group (Comp.). The levels of (A) CD28, (B) B7-1, (C) Interleukin 13 (IL-13), (D) Eotaxin-3, and (E) Tissue Inhibitor of Metalloproteinase 1 (TIMP-1). The data presented as Tukey’s boxplots of the 4 groups Neg. Ctr. (N=31), Neg. Comp. (N=17), Pos. Ctr. (N=46), and Pos. Comp. (N=16). Statistical analysis was performed using 2-way ANOVA followed by Tukey’s multiple comparisons test if the effect of alcohol and/or complication was significant, with significance indicated by *p < 0.05.

Article Snippet: ELISA kits for CD28 (#DY342-05), B7-1 (#DY140) (R&D Systems, Minneapolis, MN, USA), Interleukin 13 (IL-13; #900-K23), Eotaxin-3 (#900-K167), and Tissue Inhibitor of Metalloproteinase 1 (TIMP-1; #900-K438) (PeproTech, Hamburg, Germany) were used according to the manufacturer’s protocols.

Techniques: Enzyme-linked Immunosorbent Assay, Activation Assay, Control