Journal: Frontiers in Immunology

Article Title: Effects of Receptor Specificity and Conformational Stability of Influenza A Virus Hemagglutinin on Infection and Activation of Different Cell Types in Human PBMCs

doi: 10.3389/fimmu.2022.827760

Figure Lengend Snippet: Regulation of cytokines in primary human PBMCs during infection with HK and its HA variants. (A) mRNA expression of cytokines ( IFN-α, CCL5, IL-6 ) and nucleic acid sensor ( RIG-I ) of PBMCs inoculated at MOI 3 with HK and its HA variants HK-H17R and HK-R2 measured 6 hours p.i. using RT-PCR. Mean expression values +SD of four donors in duplicates are shown. Raw data of cytokines were normalized to the expression of the housekeeping gene RPS11 and related to HK infected samples. Significance determined by ANOVA followed by Turkey’s multiple comparison test. P values shown if significant *p < 0.05, **p < 0.01, ***p < 0.001. (B, C) MACS-isolated pDCs were either inoculated with recombinant IAVs at MOI 1 or stimulated with synthetic TLR-ligand ODN 2216 for 24 hours. Concentrations of (B) IFN-α or (C) IL-6 in the cell supernatants were determined by ELISA and depicted as mean amount of cytokine [ng/ml] +SD of two donors in duplicates.

Article Snippet: The following antibodies were used: capture: rat anti-human IL-6 (Pharmingen, 554543) or anti-human IFN-α coating antibody (Bender, 2010-10), detection: biotin rat anti-human IL-6 (Pharmingen, 554546) followed by streptavidin-POD conjugate (Roche, 11089153001) or anti-human-IFN-α HRP-conjugate (eBioscience, BM216MSTK).

Techniques: Infection, Expressing, Reverse Transcription Polymerase Chain Reaction, Isolation, Recombinant, Enzyme-linked Immunosorbent Assay

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: siRNA Library Screening Identifies a Druggable Immune-Signature Driving Esophageal Adenocarcinoma Cell Growth

doi: 10.1016/j.jcmgh.2018.01.012

Figure Lengend Snippet: LIF hierarchically regulates STAT3 phosphorylation, IL6 expression, and cell proliferation in EAC cells. SiRNA-mediated silencing of LIF expression results in reduced LIF mRNA ( A ) expression, ( B ) secretion, and ( C ) a recombinant-recoverable reduction in cell proliferation in CP-D, SKGT4, and OE-33 EAC cell lines. Treatment of LIF silenced cells with recombinant LIF protein (rLIF) results in ( D ) recovery of LIF mRNA in CP-D, SKGT4, and OE33 EAC cells and ( E ) protein expression levels in SKGT4 EAC cells. ( F ) STAT3-Y705 phosphorylation levels in LIF-silenced SKGT4 EAC cells are rescued by treatment with recombinant LIF protein as determined by Western blot. ( G ) Silencing LIF expression in SKGT4 EAC cells results in decreased IL6 mRNA expression that is recovered upon exposure to rLIF protein. ( H ) Neither silencing IL6 expression by siRNA-mediated silencing nor treatment with anti-IL6 antibody affects LIF mRNA levels or SKGT4 EAC cell growth. ( I ) Silencing LIF expression results in decreased expression of C1QA mRNA but not TREM2 in SKGT4 EAC cells. ∗ P < .05, ** P < .01, *** P < .001, and **** P < .0001 in either Student t test (proliferation) or Mann–Whitney testing (mRNA expression). siNT, nontargeting siRNA pool; siLIF, LIF-targeted siRNA pool; rLIF, recombinant LIF protein; rIL6, recombinant IL6 protein.

Article Snippet: Treatments with recombinant leukemia inhibitory factor (LIF) (30 ng/mL, ab57665; Abcam), IL6 (30 ng/mL; R&D Systems, Minneapolis, MN), native C1q (75 μg/mL; Sigma, Oakville, Ontario), and IL6 blocking antibody (AB206NA) experiments were performed in either 6- or 96-well cell culture plates for protein and viability experiments, respectively.

Techniques: Phospho-proteomics, Expressing, Recombinant, Western Blot, MANN-WHITNEY

Journal: Oncology Letters

Article Title: Significance of interstitial tumor-associated macrophages in the progression of lung adenocarcinoma

doi: 10.3892/ol.2016.5270

Figure Lengend Snippet: Typical expression images of immunochemistry staining in tissue microarrays. (A and B) Low expression of CD68 in MIA tissues. Original magnification, (A) ×100 and (B) ×200. (C and D) High expression of CD68 in LPA tissues. Original magnification, (C) ×100 and (D) ×200. (E) Negative expression of IL-6 in MIA tissues. Original magnification, ×200. (F) Positive expression of IL-6 in LPA tissues. Original magnification, ×200. (G) Negative expression of CSF-1 in MIA tissues. Original magnification, ×200. (H) Positive expression of CSF-1 in LPA tissues. Original magnification, ×200. (I) High E-cadherin expression in MIA tissues. Original magnification, ×100. (J) Low E-cadherin expression in LPA tissues. Original magnification, ×100. (K) High Snail expression in LPA tissues. Original magnification, ×100. (L) High MMP-2 expression in LPA tissues. Original magnification, ×100. CD, cluster of differentiation; MIA, minimally invasive adenocarcinoma; LPA, lepidic predominant adenocarcinoma; CSF, colony-stimulating factor; IL, interleukin; MMP, metalloproteinase.

Article Snippet: Immunohistochemistry was performed with mouse anti-human CD68 monoclonal antibody at 1:5 dilution (clone KP1; Abcam, Cambridge, UK), rabbit anti-human colony-stimulating factor (CSF)-1 polyclonal antibody at 1:100 dilution (BA0750; Wuhan Boster Biological Technology, Ltd., Wuhan, China), anti-interleukin (IL)-6 at 1:100 dilution (BS0781R; BIOSS, Beijing, China), anti-matrix metalloproteinase (MMP)-2 at 1:400 dilution (ab37150; Abcam), anti-E-cadherin at 1:200 dilution (sc-7,870; Santa Cruz Biotechnology, Inc., Dallas, TX, USA) and anti-Snail at 1:25 dilution (AP2054a; Abgent, Inc., San Diego, CA, USA).

Techniques: Expressing, Staining

Journal: Oncology Letters

Article Title: Significance of interstitial tumor-associated macrophages in the progression of lung adenocarcinoma

doi: 10.3892/ol.2016.5270

Figure Lengend Snippet: (A) Kaplan-Meier analysis of overall survival for the combination of CD68, CSF-1 and IL-6. (B) Kaplan-Meier analysis of disease-free survival for the combination of CD68, CSF-1 and IL-6. CSF-1, colony-stimulating factor-1; CD, cluster of differentiation; IL, interleukin,

Article Snippet: Immunohistochemistry was performed with mouse anti-human CD68 monoclonal antibody at 1:5 dilution (clone KP1; Abcam, Cambridge, UK), rabbit anti-human colony-stimulating factor (CSF)-1 polyclonal antibody at 1:100 dilution (BA0750; Wuhan Boster Biological Technology, Ltd., Wuhan, China), anti-interleukin (IL)-6 at 1:100 dilution (BS0781R; BIOSS, Beijing, China), anti-matrix metalloproteinase (MMP)-2 at 1:400 dilution (ab37150; Abcam), anti-E-cadherin at 1:200 dilution (sc-7,870; Santa Cruz Biotechnology, Inc., Dallas, TX, USA) and anti-Snail at 1:25 dilution (AP2054a; Abgent, Inc., San Diego, CA, USA).

Techniques:

Journal: Oncology Letters

Article Title: Significance of interstitial tumor-associated macrophages in the progression of lung adenocarcinoma

doi: 10.3892/ol.2016.5270

Figure Lengend Snippet: (A) OS in patients with AIS/MIA in the CD68+CSF-1+IL-6+ group compared with other groups, including the CD68-, CD68+CSF-1+IL-6-, CD68+CSF-1-IL-6+ and CD68+CSF-1-IL-6- groups. (B) DFS in patients with AIS/MIA in the CD68+ CSF-1+ IL-6+ group compared with other groups (C) Comparison of OS in AIS/MIA patients with CD68+CSF-1+IL-6+ and in LPA patients with CD68+CSF-1+IL-6+. (D) Comparison of DFS in AIS/MIA patients with CD68+CSF-1+IL-6+ and in LPA patients with CD68+CSF-1+IL-6+. *Others include the CD68+CSF-1+IL-6- group, the CD68+CSF-1-IL-6+ group, the CD68+CSF-1-IL-6- group and the CD68- group. AIS, adenocarcinoma in situ ; MIA, minimally invasive adenocarcinoma; CD, cluster of differentiation; CSF, colony-stimulating factor; IL, interleukin; OS, overall survival; DFS, disease-free survival.

Article Snippet: Immunohistochemistry was performed with mouse anti-human CD68 monoclonal antibody at 1:5 dilution (clone KP1; Abcam, Cambridge, UK), rabbit anti-human colony-stimulating factor (CSF)-1 polyclonal antibody at 1:100 dilution (BA0750; Wuhan Boster Biological Technology, Ltd., Wuhan, China), anti-interleukin (IL)-6 at 1:100 dilution (BS0781R; BIOSS, Beijing, China), anti-matrix metalloproteinase (MMP)-2 at 1:400 dilution (ab37150; Abcam), anti-E-cadherin at 1:200 dilution (sc-7,870; Santa Cruz Biotechnology, Inc., Dallas, TX, USA) and anti-Snail at 1:25 dilution (AP2054a; Abgent, Inc., San Diego, CA, USA).

Techniques: In Situ