Journal: Heliyon

Article Title: Cytokine expression and mortality risk among COVID-19 hospitalized patients over 60 years of age in a referral hospital in Cartagena, Colombia

doi: 10.1016/j.heliyon.2024.e29028

Figure Lengend Snippet: Biological markers and cytokine levels. Concentrations or levels are presented using median (IQR).

Article Snippet: The quantification of IL-2 (Ref: 12-7029-42), IL-4 (Ref: 48-7049-42), IL-6 (Ref: 46-7069-42), IL-10 (Ref: 53-7108-42), TNF-α (Ref: 56-7349-42), and IFN-γ (Ref: 47-7319-42) in peripheral blood leukocytes was conducted using intracellular reagents and antibodies (Invitrogen eBioscience) following manufacturer's recommendations on an Attune NxT flow cytometer (ThermoFisher; catalog number: A24861).

Techniques: Coagulation

Journal: Heliyon

Article Title: Cytokine expression and mortality risk among COVID-19 hospitalized patients over 60 years of age in a referral hospital in Cartagena, Colombia

doi: 10.1016/j.heliyon.2024.e29028

Figure Lengend Snippet: Expression levels of cytokines stratified by ICU admission and death. A) IL-2. B) IL-4. C) IL-6. D) IL-10. E) TNF-α. F) IFN-γ. Box plots show values transformed using the ln function. *p < 0.010, **p < 0.05.

Article Snippet: The quantification of IL-2 (Ref: 12-7029-42), IL-4 (Ref: 48-7049-42), IL-6 (Ref: 46-7069-42), IL-10 (Ref: 53-7108-42), TNF-α (Ref: 56-7349-42), and IFN-γ (Ref: 47-7319-42) in peripheral blood leukocytes was conducted using intracellular reagents and antibodies (Invitrogen eBioscience) following manufacturer's recommendations on an Attune NxT flow cytometer (ThermoFisher; catalog number: A24861).

Techniques: Expressing, Transformation Assay

Journal: Cells

Article Title: Peripheral Blood Mononuclear Cells from Patients with Type 1 Diabetes and Diabetic Retinopathy Produce Higher Levels of IL-17A, IL-10 and IL-6 and Lower Levels of IFN-γ—A Pilot Study

doi: 10.3390/cells12030467

Figure Lengend Snippet: Antibodies for flow cytometry and conditions for immunostaining of peripheral blood mononuclear cells (PBMC).

Article Snippet: IL-6 , MQ2-13A5 , APC , 1:40 , , eBioscience , 17-7069-42.

Techniques: Flow Cytometry, Immunostaining

Journal: Scientific Reports

Article Title: PLCγ2 regulates TREM2 signalling and integrin-mediated adhesion and migration of human iPSC-derived macrophages

doi: 10.1038/s41598-021-96144-7

Figure Lengend Snippet: Survival and phagocytosis are reduced in PLCγ2 Ko iPSC macrophages, while LPS-induced secretion of inflammatory cytokines TNFα and IL-6 is unaffected. ( A ) Cell death is slightly increased under normal culture conditions after ≥ 7 days in the PLCγ2 Ko line compared to the Parent. n = 3 ( B ) In the absence of M-CSF, PLCγ2 Ko cells show enhanced sensitivity to cell death, as the percentage of dead cells is increasing earlier than in the Parent line. n = 3 ( C ) Representative live-cell images of human iPSC macrophages phagocytosing pHrodo-labelled synaptosomes and quantification of phagocytic uptake indicated as number of spots per cell. PLCγ2 Ko cells show decreased phagocytosis rate compared to Parent. Cytochalasin D (cytoD, 10 µM) pre-incubation prevents phagocytic uptake in both lines. n = 4. Scale bar 50 μm, inset is a section of the image magnified threefold. ( D) Representative live-cell images of macrophages phagocytosing pHrodo-labelled dead SHSY5Y and quantification of phagocytic uptake indicated as number of spots per cell. PLCγ2 Ko cells show decreased phagocytosis rate compared to Parent. Cytochalasin D pre-incubation prevents phagocytic uptake in both lines. n = 4. Scale bar 50 μm, inset is a section of the image magnified threefold. ( E) Secretion of TNFα and IL-6 after 24 h stimulation with LPS is not affected in the absence of PLCγ2, n = 4. n.d. – not detectable. Data shown represent mean ± SEM, two-way ANOVA followed by Bonferroni’s multiple comparison test. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: For detection of IL-6, Greiner high-bind 96 well plates (Sigma) were coated with IL-6 antibody (Life technologies, #14–7069-81) overnight at 4 °C.

Techniques: Incubation

Journal: bioRxiv

Article Title: Altered metabolic function induced by amyloid-beta oligomers and PSEN1-mutations in iPSC-derived astrocytes

doi: 10.1101/2023.08.23.554346

Figure Lengend Snippet: Astrocytes (45 days old) were treated with 0.2, 1, 2µM Aβ oligomers. Cytokine or GFAP levels in media were measure using ELISA or by investigating intracellular accumulation using flow cytometry or ELISA. A) IL-6 levels (pg/ml) in the media were measured using ELISA or C) Flow cytometry following treatment of astrocytes with Aβ oligomers. B) IL-8 levels (pg/ml) in the media were measured using ELISA or D) Flow cytometry (fold change) following treatment of astrocytes with Aβ oligomers. E) Secreted levels of GFAP in the media or F) Cell lysates were measured using ELISA (ng/ml). Results are expressed as ± SD, n=3 P<0.05 (*), P<0.01 (**), P<0.001 (***). Comparisons between treatments were performed using ANOVA followed by Dunnet’s post-test.

Article Snippet: Astrocytes were washed a further two times in wash buffer before incubation with IL-6-PE (12-7069-82, Invitrogen) and IL-8 FITC (BH0814, BioLegend) conjugated antibodies for 30 minutes.

Techniques: Enzyme-linked Immunosorbent Assay, Flow Cytometry

Journal: bioRxiv

Article Title: Altered metabolic function induced by amyloid-beta oligomers and PSEN1-mutations in iPSC-derived astrocytes

doi: 10.1101/2023.08.23.554346

Figure Lengend Snippet: Cytokine, GFAP, 8-Isoprostane levels in media or cellular lysates were measured in control and fAD astrocytes (45 days old). A) Levels of GFAP were measure in the cell lysates or F) cell culture media using ELISA (ng/ml). C) Isoprostane levels were also measured in cellular lysates (pg/mg). Pooled control and fAD cell samples are compared. D) IL-6 and E) IL-8 levels were measured using Flow cytometry (fold change) following in control and fAD astrocytes. Results are expressed as ± SD, n=3 P<0.05 (*), P<0.01 (**), P<0.001 (***). For direct comparison between control and PSEN1, unpaired t-tests were performed. Comparisons between individual PSEN1 lines were performed using ANOVA followed by Dunnet’s post-test.

Article Snippet: Astrocytes were washed a further two times in wash buffer before incubation with IL-6-PE (12-7069-82, Invitrogen) and IL-8 FITC (BH0814, BioLegend) conjugated antibodies for 30 minutes.

Techniques: Cell Culture, Enzyme-linked Immunosorbent Assay, Flow Cytometry