Journal: Biochemistry and Biophysics Reports

Article Title: Alterations of the gut microbial community structure modulates the Th17 cells response in a rat model of asphyxial cardiac arrest

doi: 10.1016/j.bbrep.2023.101543

Figure Lengend Snippet: CA/CPR injury induced the Th17 cells response. (A) Th17 cells (CD4 was marked with green and IL-17 was marked with red) were present in the cortex of CA/CPR rat but was absent in the cortex of sham rat after 72 h following CA/CPR. (B) The level of IL-17, IL-6, and TNF-α increased in the serum of rats after 72 h following CA/CPR, compared with the rat in sham group. (C) The proportions of the Th17 cells in the PBMC were evidently enhanced from 1.71 ± 0.67% in the rats of sham group to 4.5 ± 1.2% in rats of CA/CPR group. (D)Th17 cells fluorescently labeled after microinjection in PPs were detected in the PBMC after 72 h following CA/CPR, and CFSE-positive cells derived from labeled PPs of small intestine accounted for 28.33 ± 6.18% of total Th17 cells (* P < 0.05). (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: The lymphocytes in the small intestines PPs of rats were labeled with carboxyfluorescein succinimidyl ester (CFSE, MedChemExpress, USA) by microinjection (25 μM in 2 ul of PBS per PP) after 24 h following CA/CPR.

Techniques: Labeling, Microinjection, Derivative Assay

Journal: Scandinavian journal of immunology

Article Title: Mucosal-associated invariant T cells modulate innate immune cells and inhibit colon cancer growth.

doi: 10.1111/sji.13391

Figure Lengend Snippet: FIGURE 3 Activation of MAIT cells enhance killing against colon cancer cells. (A) Experimental setup of human whole leukocyte isolation and killing assay co-culture with COLO 205. (B) Percent specific lysis of COLO 205 co-cultured with human whole leukocytes with or without 5-A-RU/MGO stimulation using a 4-h Calcein Release Assay. (C) Frequency of CFSE+ cells after overnight co-culture of human whole leukocytes with COLO 205 with or without 5-A-RU/MGO stimulation for 16 h by flow cytometry. Each dot represents a sample from an individual healthy donor and data were collected from two independent experiments. Figure created using BioRender.com. *p < 0.05 by Wilcoxon ranked test.

Article Snippet: For flow cytometry- based killing assay, 100,000 CFSE- stained (Cat. #: 6162, Immunochemistry Technologies, Davis, California, CA, USA) COLO 205 was co- cultured with 106 human whole leukocytes for 16 h and the frequency of CFSE+ cells was analysed.

Techniques: Activation Assay, Isolation, Co-Culture Assay, Lysis, Cell Culture, Release Assay, Flow Cytometry

Journal: Frontiers in Pharmacology

Article Title: Jianpi Huayu Decoction Attenuates the Immunosuppressive Status of H 22 Hepatocellular Carcinoma-Bearing Mice: By Targeting Myeloid-Derived Suppressor Cells

doi: 10.3389/fphar.2020.00016

Figure Lengend Snippet: Jianpi Huayu Decoction (JHD) weakens the immunosuppressive ability of Myeloid-derived suppressor cells (MDSCs) in vitro. (A) Relative expression of ROS in MDSCs was analyzed by flow cytometry (n = 3). Representative flow cytometry data and statistical diagram are shown. (B) JHD ameliorated the proliferation inhibitive ability of MDSCs on CD4 + T cells. Briefly, MDSCs which isolated from H 22 hepatocellular carcinoma bearing mice by MACS technique were pretreated with JHD (500μg/ml) or not. Naïve CD4 + T cells were isolated from 6-weeks BALB/c mice by Naïve CD4 + T cells Isolation Kit, and activated by plate-coated anti‐CD3 antibody (5μg/ml, Biogems) and soluble anti‐CD28 antibody (2μg/ml, Biogems) for 12 hours. Then CD4 + T cells and MDSCs were co-cultured at different ratios (CD4 + T cells: MDSCs = 1:1, 1:2) for 48h, and CD4 + T cells co-cultured with DC (6 × 10 ^5 cells/well) as positive control. Proliferation of CD4 + T cell was measured by carboxyl fluorescein diacetate succinimide (CFSE) using flow cytometry, with the gate of CD4 + T cells (n = 3). (C, D) Expression of STAT-3 and iNOS in spleen were measured by immunohistochemistry staining (×400 magnification, n = 5). Scale bar = 50μm. *: p < 0.05; **: p < 0.01; ns, not significant.

Article Snippet: The cells were then labelled with carboxyl fluorescein diacetate succinimide (CFSE) and cultured in 24-well plates that were coated with anti-CD3 (10μg/ml, BioGems) and anti-CD28 (2μg/ml, BioGems) antibodies for 12 hours.

Techniques: Derivative Assay, In Vitro, Expressing, Flow Cytometry, Isolation, Cell Culture, Positive Control, Immunohistochemistry, Staining