Journal: Clinical Cancer Research

Article Title: Autocrine 17-β-Estradiol/Estrogen Receptor-α Loop Determines the Response to Immune Checkpoint Inhibitors in Non–Small Cell Lung Cancer

doi: 10.1158/1078-0432.ccr-22-3949

Figure Lengend Snippet: Figure 1. ERa expression is a predictive factor of the response to pembrolizumab in patients with NSCLC. Best response (PD, progressive disease; PR, partial response; SD, stable disease) 6 months after the beginning of pembrolizumab treatment as first-line monotherapy, PFS, and OS (Kaplan–Meier and log-rank tests) were analyzed in 35 patients with NSCLC (15 females: F; 20 males: M). A and B, Best response,PFS, andOSin female and male patients. C and D, Best response, PFS, and OS in patients stratified according to the median values of CD274/PD-L1 mRNA measured by RT-PCR (technical triplicates) in the tumor samples. E and F, Best response, PFS, and OS in patients stratified according to the median values of ESR1/ERa mRNA measured by RT- PCR (technical triplicates) in the tumor samples. Median ERa value was calculat- ed using ERþ and ER breast cancers as highest and lowest values of the scale. G, PFS and OS in PD-L1low versus PD-L1high patients stratified according to the median values of PD-L1 protein, eval- uated by immunohistochemical analysis (TPS score, Supplementary Table S2). H, Mean PFS and OS of 6 patients (3 females and 3 males) classified as ERahigh and ERalow according to immu- nohistochemical analysis (H score; Supplementary Fig. S2B). Median ERa value was calculated using ERþ and ER

Article Snippet: ER-a overexpression and silencing NCI-H1385 cells (1 106) were transfected with 1 mg of Lenti ORF clone of Human estrogen receptor 1 (ESR1), transcript variant 1, mutant green fluorescent protein (mGFP)-tagged (#RC213277L4; Origene); 1 106 NCI-H1975 cells were transduced with 1 mg of lentiviral GFP-plasmid containing 4 unique 29mer shRNA constructs for ESR1 (#TL320346; Origene), using TurboFectin 8.0 (#TF81001; OriGene) as per the manufacturer’s instructions.

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Immunohistochemical staining

Journal: Frontiers in Microbiology

Article Title: Enterovirus 71 Antagonizes Antiviral Effects of Type III Interferon and Evades the Clearance of Intestinal Intraepithelial Lymphocytes

doi: 10.3389/fmicb.2021.806084

Figure Lengend Snippet: IFN-λ effectively inhibits EV71 infection in intestinal epithelial cells. (A) HT29 and Jurkat T cells were uninfected or infected with EV71 at an MOI of 1, qRT-PCR was used to detect transcriptional upregulation of both IFN-β, IFN-λ1, and IFN-λ2 at 48 hpi. (B) At the same infection conditions, the production and secretion of IFN-β, IFN-λ1, and IFN-λ2 proteins in the supernatant were detected by ELISA. Mock was cells without infection. (C) HT29 and Jurkat T cells were stimulated without or with 10 ng/ml recombinant protein IFN-β or IFN-λ1 for 12 h. Then the expression levels of ISG54, ISG15, PKR, and OAS were measured by qRT-PCR. Calculated the expression level of each gene relative to the expression of GAPDH and normalized it to mock-treated cells. Control was cells without treatment. (D,E) HT29 cells were pretreated with 10 ng/ml IFN-λ or 10 ng/ml IFN-β for 6 h, respectively, and then infected with EV71 at MOI of 1 for 24 h. The virus replication was monitored by qRT-PCR and plaque experiments. Control was cells with infected but without IFN treatment. Dates were presented as mean ± SD of three replicates ( n = 3 independent experiments, ** P < 0.01, *** P < 0.001, *⁣*⁣** P < 0.0001 and ns, not significant).

Article Snippet: The secretion levels of IFN-β, IFN-λ1, and IFN-λ2 proteins in the supernatant were detected by commercially available enzyme-linked immunosorbent assay kits, including Human IFN-β ELISA Kit (LIANKE, EK1236), Human IL-28A ELISA Kit (BOSTER, EK0969), and Human IL-29 ELISA Kit (BOSTER, EK0964).

Techniques: Infection, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Recombinant, Expressing, Control, Virus

Journal: Frontiers in Microbiology

Article Title: Enterovirus 71 Antagonizes Antiviral Effects of Type III Interferon and Evades the Clearance of Intestinal Intraepithelial Lymphocytes

doi: 10.3389/fmicb.2021.806084

Figure Lengend Snippet: The 2Apro and 3Cpro of EV71 antagonize the antiviral function of IFN-λ and inhibit the expression level of IFN-λ induced by poly(I:C). HT29 cells were transfected with pcDNA3.1-2A (control plasmid: pcDNA3.1) or p-EGFP-3C (control plasmid: p-EGFP). After transfection 24 h, cells were treated with IFN-λ (10 ng/ml) 6 h before infection. Then cells were infected with EV71 at an MOI of 1. Cells and culture mediums were collected at 24 hpi. (A) The protein expression level of VP1 was determined by incubating with anti-EV71 VP1 antibody, followed by staining with Alexa Fluor 555-conjugated secondary antibody. (B) The mRNA expression levels of VP1 were determined by qRT-PCR. (C) Cell culture medium was prepared for virus titer analysis. Control was cells with infected only. IFN-λ group was cells infected and using IFN-λ pretreatment. (D,E) HT29 cells were transfected with pcDNA3.1-2A (pcDNA3.1) or p-EGFP-3C (p-EGFP), and after transfection 24 h, cells were treated with poly(I:C) (4 μg/ml). Cells and culture mediums were collected 24 h after stimulation. The protein expression levels of IFN-λ1 and IFN-λ2 were determined by ELISA, and the mRNA expression levels were determined by qRT-PCR. Control was cells without any treatment. PIC group was cells stimulated only by poly(I:C). Dates were presented as mean ± SD ( n = 3 independent experiments, * P < 0.05, ** P < 0.01 and *** P < 0.001).

Article Snippet: The secretion levels of IFN-β, IFN-λ1, and IFN-λ2 proteins in the supernatant were detected by commercially available enzyme-linked immunosorbent assay kits, including Human IFN-β ELISA Kit (LIANKE, EK1236), Human IL-28A ELISA Kit (BOSTER, EK0969), and Human IL-29 ELISA Kit (BOSTER, EK0964).

Techniques: Expressing, Transfection, Control, Plasmid Preparation, Infection, Staining, Quantitative RT-PCR, Cell Culture, Virus, Enzyme-linked Immunosorbent Assay

Journal: British Journal of Cancer

Article Title: Novel urinary protein biomarker panel for early diagnosis of gastric cancer

doi: 10.1038/s41416-020-01063-5

Figure Lengend Snippet: Urinary protein level.

Article Snippet: To measure each urinary protein concentration, we used the respective ELISA kits for trefoil factor 1 (TFF1) (#EK1232; Boster Biological Technology, Pleasanton, CA), a disintegrin and metalloproteinase domain-containing protein 12 (ADAM12) (#.DAD120; R&D Systems, Minneapolis, MN), pepsinogen 3 (PGA3) (#OKEH03169; AVIVA SYSTEMS BIOLOGY, San Diego, CA), BRCA1-associated RING domain 1 (BARD1) (#.MBS7236583; MyBioSource, San Diego, CA), coiled coil domain-containing protein 38 (CCDC38) (#.MBS7209768; MyBioSource), tubulointerstitial nephritis antigen-like 1 (TINAGL1) (#MBS9340283; MyBioSource), DEAD-box helicase 55 (DDX55) (#MBS7244207; MyBioSource) and a Parameter Creatinine Assay (R&D Systems) for creatinine.

Techniques: