Journal: Journal of Medical Virology

Article Title: Preclinical Screening Platform Identifies Azatadine‐Dimaleate as a Potent Repurposed Therapeutic Against SARS‐CoV‐2 Infection

doi: 10.1002/jmv.70713

Figure Lengend Snippet: Azatadine‐Dimaleate modulates innate immune responses in WD‐PBEC cultures. RNA samples were harvested at 96 hpi from each condition in duplicate from WD‐PBECs and a panel of selected innate immune genes (IFNL1, IFNB, IFIT1, ISG15, IL‐6, CXCL8/IL8), were quantified using RT‐qPCR assay. The relative ratios for each gene were calculated in reference to a house keeping gene GAPDH using light cycler 96 software 1.1. The data value plotted for each donor in each condition is an average of 2 biological duplicate Transwells and four technical replicates. Where possible, statistical significance was determined using unpaired Student t‐test between DMSO control and individual treatments (* p ≤ 0.05, ** p ≤ 0.01).

Article Snippet: Ready‐made catalogue assays for a set of primers and probes (FAM labelled) (Thermofisher) specific for human genes involved in the innate immune response to SARS‐COV‐2 such as: IFNL1 (Assay ID: Hs00601677 g1, Cat. No:433118) IFNB (Assay ID:Hs01077958_s1, Cat. No. 4331182), ISG15 (Assay ID: Hs01921425 s1, catalogue no:4331182), IFIT1 (Hs03027069_s1, Cat no. 4331182), IL6 (Hs00174131_m1, Cat no. 4331182) and CXCL8/IL8 (Bt03211906_m1, Cat no. 4331182) were selected for analyses.

Techniques: Quantitative RT-PCR, Software, Control

Journal: Cancer Research

Article Title: Intracellular IL1α in Peritumoral Monocytes Induces IL8 Production and Inhibits Mitophagy to Promote Stemness and Metastasis of Hepatocellular Carcinoma

doi: 10.1158/0008-5472.CAN-24-3355

Figure Lengend Snippet: Upregulated IL1α induces the production of IL8 by monocytes. A–F, CD14 + cells were purified from the peripheral blood of healthy donors. Cells were transfected with siNC or si IL1A and then treated with or without TSN for 24 hours. The expression levels of pro–IL1α were determined by immunoblotting ( n = 4; A ) and flow cytometry ( n = 4; B and C ). The expression levels of PD-L1, CD80, CD86, and HLA-DR were analyzed by flow cytometry ( n = 4; D ). The levels of TNFα, IL6, IL10, IL1β, and IL8 production were measured by ELISA ( n = 5; E and F ). G, CD14 + cells were purified from paired nontumor tissues (N) and tumor tissues (T) from eight patients with HCC, and expression levels of IL8 were quantified by qPCR. H–J, CD14 + cells were purified from tumor tissues of 20 patients with HCC, and expression levels of IL8 , IL1A , TNFA , IL6 , IL10 , and IL1B were quantified by qPCR. Patients were then divided into two groups according to the median value of IL1A expression in tumor tissues, and levels of IL8 expression between the IL1A high and IL1A low groups were compared ( H ). The correlations between the levels of IL1A and IL8 , TNFA , IL6 , IL10 , or IL1B in these cells were analyzed ( I and J ). Results shown in C–H are represented as mean ± SEM. P values were obtained by two-way ANOVA ( C–F ), two-tailed Student t test ( G and H ), or Pearson correlation and linear regression analysis ( I and J ). *, P < 0.05; **, P < 0.01; ***, P < 0.001. MFI, mean fluorescence intensity.

Article Snippet: Cytokine concentrations were detected by ELISA kits according to the manufacturer’s instructions (IL1α, DY200-05, R&D Systems; DuoSet ELISA Ancillary Reagent Kit 2, DY008, R&D Systems; TNFα, 88-7346-86, eBioscience; IL6, 88-7066-88, eBioscience; IL10, 88-7106-88, eBioscience; IL1β, 88-7261-88, eBioscience; and IL8, 88-8086-88, eBioscience).

Techniques: Purification, Transfection, Expressing, Western Blot, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Two Tailed Test, Fluorescence

Journal: Cancer Research

Article Title: Intracellular IL1α in Peritumoral Monocytes Induces IL8 Production and Inhibits Mitophagy to Promote Stemness and Metastasis of Hepatocellular Carcinoma

doi: 10.1158/0008-5472.CAN-24-3355

Figure Lengend Snippet: IL8 produced by IL1α + monocytes enhances the stemness of cancer cells. A–F, HepG2 cells were left untreated or treated with rhIL8 (100 ng/mL) for 20 hours. The expression levels of stemness-associated genes were quantified by qPCR ( n = 3; A ). Cell proliferation rate ( B ), sphere formation ( C and D ), E-cadherin, N-cadherin, vimentin, and SNAI1 expression ( E ), and cell migration ( F ) were analyzed ( n = 4). G–L , CD14 + cells were purified from the peripheral blood of healthy donors. Cells were left untreated or treated with HepG2 TSN for 4 hours, washed, and cultured for another 24 hours before their supernatants were collected. HepG2 cells were then treated with CCM or TCM in the presence or absence of control IgG or anti–IL8 neutralizing antibody (10 μg/mL) for 20 hours. Cell proliferation rate ( G ), sphere formation ( H and I ), E-cadherin, N-cadherin, vimentin, and SNAI1 expression ( J ), and cell migration ( K and L ) were analyzed ( n = 4). M, CD14 + cells and cancer cells were purified from tumor tissues from 16 patients with HCC. Expression levels of IL8 in CD14 + cells and VIM and CDH1 expression in cancer cells were determined by qPCR, and the correlations between mRNA levels of IL8 and VIM or CDH1 were analyzed. Results shown in B , D , F , G , I , and L are represented as mean ± SEM. P values were obtained by paired two-tailed Student t test ( B , D , and F ), two-way ANOVA ( G , I , and L ), or Pearson correlation and linear regression analysis ( M ). *, P < 0.05; ***, P < 0.001.

Article Snippet: Cytokine concentrations were detected by ELISA kits according to the manufacturer’s instructions (IL1α, DY200-05, R&D Systems; DuoSet ELISA Ancillary Reagent Kit 2, DY008, R&D Systems; TNFα, 88-7346-86, eBioscience; IL6, 88-7066-88, eBioscience; IL10, 88-7106-88, eBioscience; IL1β, 88-7261-88, eBioscience; and IL8, 88-8086-88, eBioscience).

Techniques: Produced, Expressing, Migration, Purification, Cell Culture, Control, Two Tailed Test