Journal: Biochemical and biophysical research communications

Article Title: Histone deacetylase 8 inhibition suppresses mantle cell lymphoma viability while preserving natural killer cell function

doi: 10.1016/j.bbrc.2020.11.001

Figure Lengend Snippet: (A) Transcriptional expression of HDACs during cytokine stimulation of primary human NK cells. RNA from freshly isolated human NK cells or NK cells stimulated with IL2, IL12, and IL18 for 24 hours were assessed by RT-qPCR. The data represents the average of 3 independent experiments with the error bars representing the standard deviation. *p<0.05. (B) Immunoblot of HDAC8 protein expression in primary NK and NK cell lines as indicated. (C) Viability of primary NK cells during HDAC8 inhibition. NK cells were treated with PCI-34051, vorinostat, or LBH-589 at the indicated concentration, followed by stimulation with IL2, IL12, and IL15. Viability was assessed by CTG at the times indicated and normalized to the vehicle (DMSO) control. Data represents the average of 3 independent experiments and error bars represent the standard deviation. (D) Viability of ex vivo expanded NK cells during HDAC8 inhibition. Lymphokine activated killer cells were expanded for 9 days, rested (100U/ml IL2) or subsequently reactivated (500U/ml IL2) overnight, followed by exposure to PCI-34051 at the indicated concentration. Viability was assessed as in panel C. Data represents the average of 2 independent experiments with the error bars representing the standard deviation.

Article Snippet: Lymphokine activated killer cells were generated per manufactures protocol (#130-094-483, Miltenyl Biotec) and expanded for 9 days with 500U/mL IL2.

Techniques: Expressing, Isolation, Quantitative RT-PCR, Standard Deviation, Western Blot, Inhibition, Concentration Assay, Control, Ex Vivo

Journal: Med (New York, N.y.)

Article Title: Early T cell and binding antibody responses are associated with COVID-19 RNA vaccine efficacy onset

doi: 10.1016/j.medj.2021.04.003

Figure Lengend Snippet:

Article Snippet: No-GFP-CD16.NK-92 cell line (High affinity 176V) , ATCC , PTA-6967.

Techniques: Virus, Recombinant, Staining, Luminex, Neutralization, Generated, Software

Journal: Canadian Journal of Anaesthesia

Article Title: Effects of sevoflurane on natural killer cell-induced apoptosis of malignant tumour cells: an in vitro laboratory study

doi: 10.1007/s12630-026-03129-z

Figure Lengend Snippet: The three natural killer-cell-mediated tumour defense mechanisms. The first mechanism is degranulation with fast kinetics and a directed (cell–cell-contact-mediated) mechanism (A). The second mechanism is the induction of the cytokine-induced death receptor pathway (B). This mechanism is not directed (no cell–cell contact is required) and is promoted by TNF-α and IFN-γ and inhibited by IL-6. The third mechanism is extracellular-vesicle/exosome-mediated tumour defense which is not directed (C). Vesicles containing enzymes such as perforin and granzyme are released by NK cells and may induce distant, not directed target cell apoptosis. The experimental setup of this study is displayed in D and E. The natural killer cell line NK92 was treated with 2.2% sevoflurane for 2 hr. In most experiments, NK92 cells were co-incubated with leukemia cells (K562) as a target cell line for varying durations (between 0 and 48 hr); for the perforin measurements in the NK-cell supernatant and NK-cell cytoplasm, no co-incubation was performed. NK = natural killer; IL-6 = interleukin-6; IFN-γ = interferon-gamma; TNF-α = tumour necrosis factor-alpha; mRNA = messenger RNA

Article Snippet: Magnetic purification of NK cells was performed using the MACSxpress® Whole Blood NK Cell Isolation Kit, human (Miltenyi Biotech, Bergisch Gladbach, Germany).

Techniques: Incubation

Journal: Canadian Journal of Anaesthesia

Article Title: Effects of sevoflurane on natural killer cell-induced apoptosis of malignant tumour cells: an in vitro laboratory study

doi: 10.1007/s12630-026-03129-z

Figure Lengend Snippet: Sevoflurane did not affect natural killer cell viability. Live NK92 cells are defined as Annexin V− and FVS660− cells, determined by flow cytometry analysis. NK92 cells were exposed for 2 hr to sevoflurane (or to air only), followed by a short incubation phase of 1, 2, 4, and 6 hr (A), and a long one of 24 and 48 hr (B). All measurements in Fig. 2 were analyzed using two-way analysis of variance, and multiple comparisons were corrected using the Bonferroni method (four comparisons in A and three comparisons in B). Values represent means (standard deviations). Six independent experiments were included in this analysis. FVS = Fixable Viability Stain; NK = natural killer; Sevo = sevoflurane

Article Snippet: Magnetic purification of NK cells was performed using the MACSxpress® Whole Blood NK Cell Isolation Kit, human (Miltenyi Biotech, Bergisch Gladbach, Germany).

Techniques: Flow Cytometry, Incubation, Staining

Journal: Canadian Journal of Anaesthesia

Article Title: Effects of sevoflurane on natural killer cell-induced apoptosis of malignant tumour cells: an in vitro laboratory study

doi: 10.1007/s12630-026-03129-z

Figure Lengend Snippet: Sevoflurane did not affect natural killer cell degranulation, cytokine messenger ribonucleic acid induction, or the quantity of released extracellular vesicles. NK92 cell degranulation (A) was assessed by the release of CD107a (a marker of degranulation, analyzed by flow cytometry) and induction of IFN-γ (B), TNF-α (C), and IL-6 (D) mRNA (analyzed by real-time polymerase chain reaction) were quantified in NK92 cells with (sevoflurane [sevo]) or without (air) exposure to sevo and incubation with K562 tumour cells. Control cells (ctrl) were not exposed to tumour cells, and the positive degranulation control was NK92 cells stimulated (pos) with PMA and ionomycin. Ribonucleic acid data were normalized. The size (E) and number (F) of EVs released by NK92 cells with (sevo) and without (air) exposure to sevoflurane was quantified by nanotracking analysis. Particles were collected at 48 hr. An unpaired, two-sided Student’s t test was used to compare the air and sevoflurane groups (Fig. 4A–F). Bar graphs show means (standard deviations); box plots illustrate the median [interquartile range], and whiskers indicate min/max values. A minimum of three independent experiments were included in each of the analyses. CD = cluster of differentiation; Ctrl = negative control; IFN-γ = interferon-gamma; IL-6 = interleukin-6; mRNA = messenger ribonucleic acid; NK = natural killer; Pos = positive control; PMA = phorbol 12-myristate 13-acetate; Sevo = sevoflurane; TNF-α = tumour necrosis factor-alpha

Article Snippet: Magnetic purification of NK cells was performed using the MACSxpress® Whole Blood NK Cell Isolation Kit, human (Miltenyi Biotech, Bergisch Gladbach, Germany).

Techniques: Marker, Flow Cytometry, Real-time Polymerase Chain Reaction, Incubation, Control, Negative Control, Positive Control

Journal: Journal of Extracellular Vesicles

Article Title: GMP‐Compliant Process for the Manufacturing of an Extracellular Vesicles‐Enriched Secretome Product Derived From Cardiovascular Progenitor Cells Suitable for a Phase I Clinical Trial

doi: 10.1002/jev2.70145

Figure Lengend Snippet: EV enrichment in the final product after purification and concentration by TFF. (a) Particle size distribution. Size distribution profiles were obtained by NTA in spent medium, in CCM after clarification, in the retentate, and in the final product. Data were analysed with the NTA 3.2 software, and the mean and mode of each condition were determined. The data presented are from a single representative experiment. (b) Particle identity and tetraspanin enrichment over the manufacturing process. MFI of CD9, CD63 and CD81 obtained by MACSPlex Exosome kit are presented for spent media, CCM after clarification, retentate and final product. (c) Particle surface marker expression. An abundance of surface proteins on EV in the final product were determined by MACSPlex Exosome kit. (d) Particle concentration. The average concentration (in 10 9 particles/mL) of each sample was determined by NTA and by a correlation between NTA and MACPlex results. (e) Cryo‐EM analysis. Representative images of the final product depicting single‐ and multi‐bilayer bound vesicles. Scale bar = 100 nm. (f) ONi Super‐resolution Nanoparticle imaging. Representative images of a Triple‐Positive (TP) tetraspanin cluster identified within the final product. Individual CD9, CD63 and CD81 images and an overlay are presented. The count (%) for each cluster sub‐type (single, double or triple positive) for the final product is presented in the bar graph.

Article Snippet: A bead‐based multiplex immunoassay, MACSPlex Cytotoxic T/NK Cell Kit (Miltenyi Biotec, #130‐125‐800), was used to measure the levels of multiple cytokines, including Interferon‐gamma (IFN‐γ) and Interleukine‐2 (IL‐2).

Techniques: Purification, Concentration Assay, Clarification Assay, Software, Marker, Expressing, Cryo-EM Sample Prep, Imaging

Journal: Journal of Extracellular Vesicles

Article Title: GMP‐Compliant Process for the Manufacturing of an Extracellular Vesicles‐Enriched Secretome Product Derived From Cardiovascular Progenitor Cells Suitable for a Phase I Clinical Trial

doi: 10.1002/jev2.70145

Figure Lengend Snippet: Long term stability (up to 36 months of storage). (a) Particle size distribution with mean and mode particle size in the final product, by NTA, over time. (b) Tetraspanin marker expression (CD81, CD63 and CD9) in final product by MACSPlex Exosome kit, over time. (c) HUVEC proliferation assay results for final product, over time.

Article Snippet: A bead‐based multiplex immunoassay, MACSPlex Cytotoxic T/NK Cell Kit (Miltenyi Biotec, #130‐125‐800), was used to measure the levels of multiple cytokines, including Interferon‐gamma (IFN‐γ) and Interleukine‐2 (IL‐2).

Techniques: Marker, Expressing, Proliferation Assay

Journal: Journal of Extracellular Vesicles

Article Title: GMP‐Compliant Process for the Manufacturing of an Extracellular Vesicles‐Enriched Secretome Product Derived From Cardiovascular Progenitor Cells Suitable for a Phase I Clinical Trial

doi: 10.1002/jev2.70145

Figure Lengend Snippet: Summary of the components and biological effects of the EV‐enriched secretome final product. (a) Some of the protein and molecular components identified in the final product. Image created with BioRender.com . (b) Summary of cell surface markers identified by the MACSPlex Exosome kit in the final product, which their known roles in biological processes. (c) Functional effects of the final product. Image created with BioRender.com .

Article Snippet: A bead‐based multiplex immunoassay, MACSPlex Cytotoxic T/NK Cell Kit (Miltenyi Biotec, #130‐125‐800), was used to measure the levels of multiple cytokines, including Interferon‐gamma (IFN‐γ) and Interleukine‐2 (IL‐2).

Techniques: Functional Assay