Journal: PLoS pathogens

Article Title: TGF-β1 down-regulation of NKG2D/DAP10 and 2B4/SAP expression on human NK cells contributes to HBV persistence.

doi: 10.1371/journal.ppat.1002594

Figure Lengend Snippet: Figure 2. Deficient NK cell function in IT patients. (A) Deficient NK cell cytotoxicity from IT patients. NK cell cytotoxicity was assessed in primary NK cells from IT, IA, and IN patients and healthy controls using a 51Cr-release assay with K562 target cells. (B) Results show the mean 6 SEM values in 19 patients and 5 controls. Values for IT patients were less than controls (P= 0.0384). (C) Positive correlation between NK cell cytotoxicity and the percentage of NK cells expressing NKG2D and 2B4. (D) Fresh PBMCs were stimulated with/without IL-12 as described in the Materials and Methods. After 16 h, IFN-c production was determined using flow cytometry by gating on CD32CD56+ NK cells. A representative dot plot displaying intracellular IFNc staining in NK cells with the subsets indicated is shown. (E) Cumulative data are shown. (F, G) Fresh PBMCs from patients before treatment (black) or after treatment (white) were stimulated with IL-12 for 16 h. IFNc (F) and CD107a (G) production was determined using flow cytometry by gating on CD32CD56+ NK cells. (H) Induction of ex vivo NK cell cytotoxic activity after in vivo administration of antiviral treatment with nucleoside analogues until ALT reached normal levels. NK cytotoxicity was determined ex vivo by measuring the lysis of 51Cr-labelled target cells before treatment (red) or after treatment (green). Cumulative data are shown. doi:10.1371/journal.ppat.1002594.g002

Article Snippet: Flow cytometric analysis Fresh human peripheral blood mononuclear cells (PBMCs) were isolated from peripheral blood by Ficoll-Isopaque (Solarbio, China) gradient centrifugation.

Techniques: Cell Function Assay, Release Assay, Expressing, Flow Cytometry, Staining, Ex Vivo, Activity Assay, In Vivo, Analogues, Lysis

Journal: Immunity, Inflammation and Disease

Article Title: Activated Peripheral CD8 + T Lymphocytes Inhibit the Proliferation of Hippocampal Neural Stem Cells via the IFN‐γ/JAK/STAT Signaling Pathway

doi: 10.1002/iid3.70287

Figure Lengend Snippet: Isolation culture and identification of NSCs and CD8 + T cells. (A) Immunofluorescence image of NSCs, Nestin in red and DAPI in blue, scale bar = 100 μm; (B) Flow cytometry showed that more than 95% of the isolated and extracted cells were Nestin positive; (C) Flow cytometry showed that more than 95% of the isolated and extracted peripheral T lymphocytes were CD8 + T lymphocytes.

Article Snippet: Peripheral blood lymphocytes were separated with a Mouse Peripheral Lymphocytes Isolation Liquid Kit (Solarbio, China).

Techniques: Isolation, Immunofluorescence, Flow Cytometry

Journal: iScience

Article Title: Neutrophil-mediated delivery of the combination of colistin and azithromycin for the treatment of bacterial infection.

doi: 10.1016/j.isci.2022.105035

Figure Lengend Snippet: Figure 1. Schematic illustration of azithromycin and colistin loaded neutrophils for the effective treatment of bacterial infection in a mouse acute pneumonia model Azithromycin is taken up directly by neutrophils. Intracellular delivery of colistin is mediated by colistin loaded liposomes. Mice are infected by P. aeruginosa intranasally and neutrophils are administered through tail vein injection. AZM, azithromycin; CST, colistin.

Article Snippet: Mouse neutrophils were isolated from bone marrow by a mouse bone marrow neutrophil isolation kit (Solarbio, Beijing, China).

Techniques: Infection, Liposomes, Injection

Journal: iScience

Article Title: Neutrophil-mediated delivery of the combination of colistin and azithromycin for the treatment of bacterial infection.

doi: 10.1016/j.isci.2022.105035

Figure Lengend Snippet: Figure 2. Uptake of azithromycin by neutrophils and the bacteria-killing efficacies of the neutrophils (A) Schematic illustration of the isolation of neutrophils and uptake of azithromycin by neutrophils. (B) Uptake kinetics of azithromycin by neutrophils. The intracellular amounts of azithromycin were determined by ELISA. Results represent means G SD. (C) Release of azithromycin after neutrophils were incubated with 100 mg/mL azithromycin for 2 h. The extracellular concentrations of azithromycin were determined by ELISA. Results represent means G SD. (D) Time-kill curves of neutrophils against PA14. Neutrophils were incubated in RPMI with 20% FBS in the absence or presence of 100 mg/mL azithromycin for 2 h. After resuspension with RPMI, the neutrophils at the concentration of 23106 cells/mL were incubated with 2 3 107 CFU/mL PA14 (total volume = 1 mL). Meanwhile, the same amount of PA14 cells was incubated with or without 2 mg/mL azithromycin in RPMI. At indicated time points, the live bacteria numbers were determined by plating. Results represent means G SD, **, p < 0.01; ***, p < 0.001 by Student’s t test.

Article Snippet: Mouse neutrophils were isolated from bone marrow by a mouse bone marrow neutrophil isolation kit (Solarbio, Beijing, China).

Techniques: Bacteria, Isolation, Enzyme-linked Immunosorbent Assay, Incubation, Concentration Assay

Journal: iScience

Article Title: Neutrophil-mediated delivery of the combination of colistin and azithromycin for the treatment of bacterial infection.

doi: 10.1016/j.isci.2022.105035

Figure Lengend Snippet: Figure 5. Effects of the intracellular azithromycin and colistin on neutrophil’s responses to bacteria (A) ROS production by neutrophils. Neutrophils loaded with azithromycin, colistin, or both drugs were incubated with PA14 (neutrophils:bacteria = 1:10). The production of ROS was monitored every 5 min for 1 h. Results represent means G SD. (B) Release of colistin and azithromycin from drug-loaded neutrophils in the presence of PA14. The extracellular drug concentrations were determined by ELISA. The cell survival percentages were determined by trypan blue staining. Results represent means G SD.

Article Snippet: Mouse neutrophils were isolated from bone marrow by a mouse bone marrow neutrophil isolation kit (Solarbio, Beijing, China).

Techniques: Bacteria, Incubation, Enzyme-linked Immunosorbent Assay, Staining