recombinant cytokines  (R&D Systems)

Journal: Cell Reports Medicine

Article Title: Development of a chimeric cytokine receptor that captures IL-6 and enhances the antitumor response of CAR-T cells

doi: 10.1016/j.xcrm.2024.101526

Figure Lengend Snippet: Attenuated Akt signaling in G6/7R-M452L supports long-term proliferation of anti-CD19 CAR-T cells (A) A schematic diagram of the individual mutants in G6/7R. (B) T cells transduced with anti-CD19 CAR alone or with anti-CD19 CAR and G6/7R with the indicated mutations were rested overnight in cytokine-free media. Immunoblotting evaluated the phosphorylated and total levels of STAT3, STAT5, and Akt. Representative data of two experiments. (C) Anti-CD19 CAR or anti-CD19 CAR+G6/7R T cells with the indicated mutations were stimulated with NALM-6 and cultured without exogenous cytokines. The data shown are fold expansion of the CAR-T cells on day 7 following stimulation ( n = 4 cultures, one-way ANOVA with multiple comparison test). Representative data of three independent experiments. (D–H) Anti-CD19 CAR-T cells with or without ectopic expression of G6/7R or G6/7R-M452L were restimulated with NALM-6. The data shown are the frequency (D) and fold expansion (E) of CD8 + central memory T cells on day 7 ( n = 4 different donor samples, repeated measures one-way ANOVA with multiple comparison test), cumulative fold expansion of CAR-T cells upon weekly stimulation until week 6 (F, n = 4 cultures, one-way ANOVA with multiple comparison test), the frequency of Annexin V + cells on day 7 (G, n = 3 cultures, one-way ANOVA with multiple-comparison test), and mean fluorescence intensity of CFSE on day 7 (H, n = 3 cultures, one-way ANOVA with multiple-comparison test). In (H), T cells were labeled with CFSE before stimulation. NS, not significant. (I–K) Mass cytometry analysis was performed in G6/7R and G6/7R-M452L anti-CD19 CAR-T cells 7 days after restimulation. (I) An EmbedSOM-based plot of concatenated samples to visualize the distribution of G6/7R and G6/7R-M452L CD8 + CAR-T cells. Individual cells are colored according to the clusters obtained by FlowSOM. Each sample consists of a pool of three cultures from one donor. (J) Proportions of each cluster in G6/7R and G6/7R-M452L CAR-T cells. Color annotations are provided for the four major populations (clusters 1, 3, 5, and 6). (K) Distribution of fluorescence intensity of the indicated molecules in each cluster. Median values with interquartile range are shown. In (C) and (F–H), horizontal lines denote mean values. See also Figure S3 and Table S2 .

Article Snippet: Extracted lysates and standard IL-6 were denatured at 95°C for 5 min. Recombinant cytokines (human IL-6 and IL-1β) were purchased from Peprotech.

Techniques: Transduction, Western Blot, Cell Culture, Comparison, Expressing, Fluorescence, Labeling, Mass Cytometry

Journal: Cellular and molecular gastroenterology and hepatology

Article Title: Oncostatin M Induces IFITM1 Expression to Inhibit Hepatitis B Virus Replication Via JAK-STAT Signaling.

doi: 10.1016/j.jcmgh.2023.10.003

Figure Lengend Snippet: Figure 1. OSM is the most effective among the examined IL6 family cytokines for suppression of HBV replication. (A) HepG2.2.15, (B) Huh7-1.3, and (C) HepAD38 cells were treated with IL6, LIF, OSM, CNTF, cardiotrophin-1 (CT-1), IL11, or IL31 at a concentration of 20 ng/mL for 72 hours. The levels of extracellular HBsAg, HBeAg, and HBV DNA were quantitated. One- way analysis of variance. (D) Quantitation of OSM concentration by enzyme-linked immunosorbent assay in serum from health control subjects and patients with chronic HBV infection in different phases. Kruskal-Wallis test. (E) Serum OSM, HBsAg, and HBeAg levels in HBeAg-positive CHB patients. Mann-Whitney. HC, health control; I, HBeAg-positive chronic HBV infection; II, HBeAg-positive CHB; III, HBeAg-negative chronic HBV infection; IV, HBeAg-negative CHB. *P < .05, **P < .01, and ***P < .001. PBS, phosphate-buffered saline.

Article Snippet: The recombinant human cytokines IL6, IL11, IL31, IL35, LIF, OSM, CNTF, and cardiotrophin-1 were purchased from R&D Systems and Novoprotein (Shanghai, China).

Techniques: Concentration Assay, Quantitation Assay, Enzyme-linked Immunosorbent Assay, Control, Infection, MANN-WHITNEY, Saline