cxcl8 Search Results


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  • 99
    Thermo Fisher gene exp cxcl8 hs00174103 m1
    Gene Exp Cxcl8 Hs00174103 M1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1575 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    R&D Systems recombinant human il 8
    Therapeutic effects of miR-200 in a basal-like breast cancer model ( a ) Relative expression levels of miR-141, <t>IL-8</t> and CXCL1 in breast cell lines. Data are averages ± s.e.m., n = 3. ( b ) Schematic of basal-like experiment (top) and primary tumour volumes (bottom) following systemic miRNA delivery. Data are averages ± s.e.m., n = 10. P -values obtained with Student's t -test. ( c ) Relative expression level of IL-8 and CXCL1 in resected primary tumours ( n = 4) from each treatment group. ( d ) Haematoxylin and eosin image of lung micrometastasis (top) and frequencies of peritoneal and microscopic lung metastases (bottom). P -values obtained with the χ 2 -test. * P
    Recombinant Human Il 8, supplied by R&D Systems, used in various techniques. Bioz Stars score: 99/100, based on 297 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    cxcl8  (Abcam)
    93
    Abcam cxcl8
    Immunohistochemical staining of <t>CXCL8</t> in cervical cancer tissues ( A ) Negative expression of CXCL8 in normal cervical tissue; ( B ) weak expression of CXCL8 in normal cervical tissue; ( C ) moderate expression of CXCL8 in normal cervical tissue; ( D ) strong expression of CXCL8 in normal cervical tissue; ( E ) negative expression of CXCL8 in cervical squamous cell carcinoma tissue; ( F ) weak expression of CXCL8 in cervical squamous cell carcinoma tissue; ( G ) moderate expression of CXCL8 in cervical squamous cell carcinoma tissue; ( H ) strong expression of CXCL8 in cervical squamous cell carcinoma tissue; ( I ) negative expression of CXCL8 in cervical adenocarcinoma tissue; ( J ) weak expression of CXCL8 in cervical adenocarcinoma tissue; ( K ) moderate expression of CXCL8 in cervical adenocarcinoma tissue; ( L ) strong expression of CXCL8 in cervical adenocarcinoma tissue.
    Cxcl8, supplied by Abcam, used in various techniques. Bioz Stars score: 93/100, based on 78 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Becton Dickinson cxcl8
    <t>CXCL8</t> + naïve T cells are enriched in CD31 + T cells. (A–D) Fresh CXCL8 + umbilical cord blood naïve T cells were analyzed for expression of surface and intracellular marks of activation and differentiation. Representative FACS plots and percentages are shown. Gated on CXCL8 + naïve T cells. (E) Graphical summary of CXCL8 expression in CD31 + and CD31 − T cells (E). n = 4–8, *, P
    Cxcl8, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 93/100, based on 127 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    PeproTech recombinant human il 8
    Tumor cell viability in human ovarian tumor cell suspensions following treatment with oncolytic adenovirus, recombinant <t>IL-8</t> or anti-IL-8 antibody Cell viability was measured using a MTS assay on days 7-13 after the start of incubation. Viability is presented as percentage of mock group cell viability. Ad = Ad5/3-d24. aIL8=anti-IL-8=anti-IL-8 neutralizing antibody. rIL8=IL-8 = recombinant IL-8. Asterisks indicate the significance of findings: * (p
    Recombinant Human Il 8, supplied by PeproTech, used in various techniques. Bioz Stars score: 94/100, based on 118 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Thermo Fisher gene exp cxcl8 hs99999034 m1
    Tumor cell viability in human ovarian tumor cell suspensions following treatment with oncolytic adenovirus, recombinant <t>IL-8</t> or anti-IL-8 antibody Cell viability was measured using a MTS assay on days 7-13 after the start of incubation. Viability is presented as percentage of mock group cell viability. Ad = Ad5/3-d24. aIL8=anti-IL-8=anti-IL-8 neutralizing antibody. rIL8=IL-8 = recombinant IL-8. Asterisks indicate the significance of findings: * (p
    Gene Exp Cxcl8 Hs99999034 M1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 92/100, based on 201 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    R&D Systems human il 8 cxcl8 antibody
    Tumor cell viability in human ovarian tumor cell suspensions following treatment with oncolytic adenovirus, recombinant <t>IL-8</t> or anti-IL-8 antibody Cell viability was measured using a MTS assay on days 7-13 after the start of incubation. Viability is presented as percentage of mock group cell viability. Ad = Ad5/3-d24. aIL8=anti-IL-8=anti-IL-8 neutralizing antibody. rIL8=IL-8 = recombinant IL-8. Asterisks indicate the significance of findings: * (p
    Human Il 8 Cxcl8 Antibody, supplied by R&D Systems, used in various techniques. Bioz Stars score: 90/100, based on 75 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    Thermo Fisher cxcl8
    PM2.5 exposure induced upregulation of VEGFA production in human bronchial epithelial cells. (A) Beas-2B cells were left untreated or were treated with different concentrations of PM2.5 (12.5, 25, 50, and 100 μg/mL) for 24 h. The production of VEGFA, IL1B, IL6, <t>CXCL8</t> and TNF was then detected in the cell culture supernatant using ELISA (*, P
    Cxcl8, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 95/100, based on 121 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Mab Technologies cxcl8
    Staining human neutrophils by anti-IL-17A (AF-317-NA) polyclonal antibodies (Abs). (A) Immunofluorescence (top panels) and immunohistochemistry (lower panels) stainings of two FFPE cases of human pustular psoriasis using anti-IL-17A (AF-317-NA) and anti-CD66b Abs (as labeled). Top panels show DAPI, FITC channel, and merge to recognize neutrophil shape; lower panels show different magnification of IHC and double IHC to characterize IL-17A + cells with the neutrophil marker CD66b. (B) Cytospins of neutrophils, either untreated (top panels) or treated with 5 µM R848 (bottom panels) for 3 h, were stained with anti-IL-17A (AF-317-NA, left panels) and <t>anti-CXCL8</t> (right panels) Abs. Original magnification 200× [first row in (A) and left image in third row, scale bar 100 µm] and 400× [second row in (A) , center/right images in third row in (A) , as well as in (B) , scale bar 50 µm]. Images of the second row in (A) represent magnifications of images in first row. (C) AF-317-NA immunoblot of lysates from neutrophils either freshly isolated (T 0 , from two donors) or incubated for 3 h with or without 2 µg/ml IL-6 plus 0.2 µg/ml IL-23 (low), 20 µg/ml IL-6 plus 2 µg/ml IL-23 (high), or 5 µM R848. Recombinant human IL-17A (rhIL-17A) was used as positive control. Panels (B,C) display representative experiments out of two independent ones with similar results.
    Cxcl8, supplied by Mab Technologies, used in various techniques. Bioz Stars score: 90/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Sanquin cxcl8
    Secretion of inflammation factors by dermal- and adipose-endothelial cells. Secretion of <t>CXCL8,</t> IL-6 and CCL2 after a 24 h exposure to 0, 40 or 100 µg/mL BWE. Basal amounts of protein in culture medium containing 100 µg/mL BWE without cells: CXCL8:
    Cxcl8, supplied by Sanquin, used in various techniques. Bioz Stars score: 90/100, based on 19 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    R&D Systems human il 8 cxcl8 biotinylated antibody
    Secretion of inflammation factors by dermal- and adipose-endothelial cells. Secretion of <t>CXCL8,</t> IL-6 and CCL2 after a 24 h exposure to 0, 40 or 100 µg/mL BWE. Basal amounts of protein in culture medium containing 100 µg/mL BWE without cells: CXCL8:
    Human Il 8 Cxcl8 Biotinylated Antibody, supplied by R&D Systems, used in various techniques. Bioz Stars score: 95/100, based on 50 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    R&D Systems anti il 8 antibody
    Alleviation of resistance via blockade of <t>IL-8</t> signaling. Cytotoxicity of PC9 and HCC827 cells mediated by NK cells or TRAIL, as indicated. Tumor cells were left untreated, treated with erlotinib for 3 days, or treated with erlotinib for 3 days followed by exposure to anti-IL-8 neutralizing antibody before the assay
    Anti Il 8 Antibody, supplied by R&D Systems, used in various techniques. Bioz Stars score: 94/100, based on 41 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    R&D Systems recombinant human il 8 cxcl8 protein
    Alleviation of resistance via blockade of <t>IL-8</t> signaling. Cytotoxicity of PC9 and HCC827 cells mediated by NK cells or TRAIL, as indicated. Tumor cells were left untreated, treated with erlotinib for 3 days, or treated with erlotinib for 3 days followed by exposure to anti-IL-8 neutralizing antibody before the assay
    Recombinant Human Il 8 Cxcl8 Protein, supplied by R&D Systems, used in various techniques. Bioz Stars score: 94/100, based on 48 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Santa Cruz Biotechnology cxcl8
    Human melanoma tumors express ligands for CXCR2, but tumor-infiltrating T cells lack CXCR2 expression. (A) Paraffin-embedded, melanoma lymph node metastases were analyzed by immunohistochemical staining for chemokine <t>CXCL8</t> and CXCL1. Representative staining is shown at 40X magnification. (B) CXCR2 expression on PBMCs, as determined by flow cytometry. PBMCs from five healthy donors were stained with anti-CD3 and anti-CXCR2. (C) FACS analysis of CXCR2 expression on tumor-infiltrating lymphocytes (TILs). TILs isolated from four melanoma patients were stained with anti-CD4, CD8 and CXCR2. Representative results of all stained samples were shown.
    Cxcl8, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 92/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Becton Dickinson cxcl8 il
    Serial changes in plasma cytokine/chemokine concentrations during the course of hospitalization. There was sustained elevation of the proinflammatory cytokines (IL-6, <t>CXCL8/IL-8,</t> CCL2/MCP-1, sTNFR-1) in severe pH1N1 pneumonia; the adaptive-immunity related cytokines (CXCL10/IP-10, CXCL9/MIG, IL-17A) were markedly suppressed compared with seasonal influenza. All patients with pH1N1 influenza (severe pneumonia, n = 34; milder illness, n = 29) received antiviral treatment soon after hospitalization/recruitment; none had received high-dose corticosteroids or other immunosuppressants for ‘viral pneumonitis’ or ‘ARDS’ [8] . Among seasonal influenza patients (most had complicated illnesses, see Table 1 footnotes), 30(57%) received antiviral treatment. Median concentrations at each time point are shown for each group; the interquartile ranges (presented in Table 1 ) are omitted here for clarity. Fewer mild pH1N1 and untreated seasonal influenza patients remained hospitalized at day 6–7 for study (Day 1, n = 116; Day 3–4, n = 62; Day 6–7, n = 30).
    Cxcl8 Il, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 90/100, based on 23 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher gene exp cxcl8 hs01553824 g1
    Serial changes in plasma cytokine/chemokine concentrations during the course of hospitalization. There was sustained elevation of the proinflammatory cytokines (IL-6, <t>CXCL8/IL-8,</t> CCL2/MCP-1, sTNFR-1) in severe pH1N1 pneumonia; the adaptive-immunity related cytokines (CXCL10/IP-10, CXCL9/MIG, IL-17A) were markedly suppressed compared with seasonal influenza. All patients with pH1N1 influenza (severe pneumonia, n = 34; milder illness, n = 29) received antiviral treatment soon after hospitalization/recruitment; none had received high-dose corticosteroids or other immunosuppressants for ‘viral pneumonitis’ or ‘ARDS’ [8] . Among seasonal influenza patients (most had complicated illnesses, see Table 1 footnotes), 30(57%) received antiviral treatment. Median concentrations at each time point are shown for each group; the interquartile ranges (presented in Table 1 ) are omitted here for clarity. Fewer mild pH1N1 and untreated seasonal influenza patients remained hospitalized at day 6–7 for study (Day 1, n = 116; Day 3–4, n = 62; Day 6–7, n = 30).
    Gene Exp Cxcl8 Hs01553824 G1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 86/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Becton Dickinson anti cxcl8 antibodies
    <t>CXCL8</t> expression via NF-κB activation in SL-treated THP-1 cells. THP-1 cells were pretreated with α2,3- or α2,6-SL (1 or 10 μM) for 3 h and then treated with LPS (0.1 μg/ml) or PAK (MOI of 50). (A) CXCL8 mRNA levels were analyzed by RT-PCR. (B and E) CXCL8 secretion was analyzed by ELISA. (C) IκBα degradation was analyzed by Western blotting. (D) THP-1 cells were pretreated with the NF-κB inhibitor BAY11-7082 (10 μM) and then treated with α2,3- or α2,6-SL (1 or 10 μM) and PAK (MOI of 50) as previously described. CXCL8 expression was analyzed by RT-PCR or ELISA. Statistical significance was determined by ANOVA (Tukey’s test). ANOVA test results are shown as letters above columns. Means not sharing the same letter are significantly different. GAPDH, glyceraldehyde 3-phosphate dehydrogenase.
    Anti Cxcl8 Antibodies, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 93/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Abcam anti il8 antibody
    <t>CXCL8</t> expression via NF-κB activation in SL-treated THP-1 cells. THP-1 cells were pretreated with α2,3- or α2,6-SL (1 or 10 μM) for 3 h and then treated with LPS (0.1 μg/ml) or PAK (MOI of 50). (A) CXCL8 mRNA levels were analyzed by RT-PCR. (B and E) CXCL8 secretion was analyzed by ELISA. (C) IκBα degradation was analyzed by Western blotting. (D) THP-1 cells were pretreated with the NF-κB inhibitor BAY11-7082 (10 μM) and then treated with α2,3- or α2,6-SL (1 or 10 μM) and PAK (MOI of 50) as previously described. CXCL8 expression was analyzed by RT-PCR or ELISA. Statistical significance was determined by ANOVA (Tukey’s test). ANOVA test results are shown as letters above columns. Means not sharing the same letter are significantly different. GAPDH, glyceraldehyde 3-phosphate dehydrogenase.
    Anti Il8 Antibody, supplied by Abcam, used in various techniques. Bioz Stars score: 97/100, based on 29 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    R&D Systems recombinant human cxcl8 il
    Effect of nitric oxide addition on binding and dissociation of the LDV-FITC probe on U937 cells, treated with different Gα i -coupled receptor ligands . LDV-FITC probe binding and dissociation on U937 cells stably transfected with different GPCRs plotted as mean channel fluorescence (MCF) versus time. A, The experiment involved sequential addition of fluorescent LDV-FITC probe (4 nM, below saturation, added 2 min prior to addition of Gαi-coupled receptor ligand, fMLFF, 100 nM), and different concentrations of DEA-NONOate (nitric oxide donor). Dashed line indicates the non-specific binding of the LDV-FITC probe determined using an excess of unlabelled LDV competitor (as shown in Fig. 2D,E). B, The experiment involved sequential addition of fluorescent LDV-FITC probe (4 nM), CXCL12/SDF-1 (12 nM), and DEA-NONOate (250 μM, nitric oxide donor) or vehicle (control). Rapid and reversible binding of the probe reflects the VLA-4 affinity change [14]. C, The experiment involved sequential addition of the fluorescent LDV-FITC probe (4 nM), <t>CXCL8/IL-8</t> (20 nM), and DEA-NONOate (250 μM, nitric oxide donor) or vehicle (control). D, The experiment involved sequential addition of the DEA-NONOate (250 μM, nitric oxide donor) or vehicle (control) at the 0 time point, and the fluorescent LDV-FITC probe (4 nM), CXCL12/SDF-1 (12 nM). Excess unlabelled competitor LDV (1 μM) is added at the end of the experiment to determine the non-specific binding of the probe (panels D, and E). E, The experiment involved sequential addition of DEA-NONOate (250 μM, nitric oxide donor) or vehicle (control) at the 0 time point, and the fluorescent LDV-FITC probe (4 nM), CXCL8/IL-8 (20 nM) (arrows). According to the unpaired t test, the means are significantly different (p
    Recombinant Human Cxcl8 Il, supplied by R&D Systems, used in various techniques. Bioz Stars score: 93/100, based on 13 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Therapeutic effects of miR-200 in a basal-like breast cancer model ( a ) Relative expression levels of miR-141, IL-8 and CXCL1 in breast cell lines. Data are averages ± s.e.m., n = 3. ( b ) Schematic of basal-like experiment (top) and primary tumour volumes (bottom) following systemic miRNA delivery. Data are averages ± s.e.m., n = 10. P -values obtained with Student's t -test. ( c ) Relative expression level of IL-8 and CXCL1 in resected primary tumours ( n = 4) from each treatment group. ( d ) Haematoxylin and eosin image of lung micrometastasis (top) and frequencies of peritoneal and microscopic lung metastases (bottom). P -values obtained with the χ 2 -test. * P

    Journal: Nature communications

    Article Title: Tumour angiogenesis regulation by the miR-200 family

    doi: 10.1038/ncomms3427

    Figure Lengend Snippet: Therapeutic effects of miR-200 in a basal-like breast cancer model ( a ) Relative expression levels of miR-141, IL-8 and CXCL1 in breast cell lines. Data are averages ± s.e.m., n = 3. ( b ) Schematic of basal-like experiment (top) and primary tumour volumes (bottom) following systemic miRNA delivery. Data are averages ± s.e.m., n = 10. P -values obtained with Student's t -test. ( c ) Relative expression level of IL-8 and CXCL1 in resected primary tumours ( n = 4) from each treatment group. ( d ) Haematoxylin and eosin image of lung micrometastasis (top) and frequencies of peritoneal and microscopic lung metastases (bottom). P -values obtained with the χ 2 -test. * P

    Article Snippet: For rescue experiments (RF24 cells), 10 ng ml−1 of recombinant human IL-8 (R & D Systems, Minneapolis, MN) was added to complete media as a chemo-attractant.

    Techniques: Expressing

    Therapeutic effects of miR-200 delivery in metastatic models ( a ) Average mass of the left lung (left) and number of lung nodules (right) in the 344SQ lung cancer model. Data are averages ± s.e.m., n = 10. P -values obtained with Student's t -test. ( b ) Average aggregate mass of distant metastases (left) and number of distant metastases (right), n = 10 per group. Data are averages ± s.e.m. P -values were obtained with Student's t -test. ( c ) Representative images of an orthotopic left lung tumour (left, encircled), mediastinal (middle, arrows) and liver metastases (right, arrows). ( d ) Frequency of lung cancer metastases. P -values obtained with χ 2 -test. ( e ) Aggregate mass of distant metastases (left) and number of metastases (right) in the HeyA8 ovarian cancer model, n = 10 per group, Data are averages ± s.e.m. ( f ) Frequency of metastases to distant sites. Other: the pelvis, ovary, kidney or liver. ( g ) Representative tumour staining of IL-8 in the HeyA8 model. Scale bar, 200 μm. ( h ) Microvessel density for HeyA8 tumours following miRNA delivery, n = 5 tumours per group. Data are averages ± s.e.m. P -values obtained with Student's t -test. Scale bar, 200 μm. ( i ) Representative images of gross in vivo (top, white arrows) and aggregate ex vivo (bottom) metastatic tumour burden in the HeyA8 ovarian model at the time of necropsy. ( j ) Aggregate mass of metastasis ( n = 10 mice per group) and ( k ) IL-8 plasma levels ( n = 5 per group) in the HeyA8 model. Data are averages ± s.e.m. P -values obtained with Student's t -test. * P

    Journal: Nature communications

    Article Title: Tumour angiogenesis regulation by the miR-200 family

    doi: 10.1038/ncomms3427

    Figure Lengend Snippet: Therapeutic effects of miR-200 delivery in metastatic models ( a ) Average mass of the left lung (left) and number of lung nodules (right) in the 344SQ lung cancer model. Data are averages ± s.e.m., n = 10. P -values obtained with Student's t -test. ( b ) Average aggregate mass of distant metastases (left) and number of distant metastases (right), n = 10 per group. Data are averages ± s.e.m. P -values were obtained with Student's t -test. ( c ) Representative images of an orthotopic left lung tumour (left, encircled), mediastinal (middle, arrows) and liver metastases (right, arrows). ( d ) Frequency of lung cancer metastases. P -values obtained with χ 2 -test. ( e ) Aggregate mass of distant metastases (left) and number of metastases (right) in the HeyA8 ovarian cancer model, n = 10 per group, Data are averages ± s.e.m. ( f ) Frequency of metastases to distant sites. Other: the pelvis, ovary, kidney or liver. ( g ) Representative tumour staining of IL-8 in the HeyA8 model. Scale bar, 200 μm. ( h ) Microvessel density for HeyA8 tumours following miRNA delivery, n = 5 tumours per group. Data are averages ± s.e.m. P -values obtained with Student's t -test. Scale bar, 200 μm. ( i ) Representative images of gross in vivo (top, white arrows) and aggregate ex vivo (bottom) metastatic tumour burden in the HeyA8 ovarian model at the time of necropsy. ( j ) Aggregate mass of metastasis ( n = 10 mice per group) and ( k ) IL-8 plasma levels ( n = 5 per group) in the HeyA8 model. Data are averages ± s.e.m. P -values obtained with Student's t -test. * P

    Article Snippet: For rescue experiments (RF24 cells), 10 ng ml−1 of recombinant human IL-8 (R & D Systems, Minneapolis, MN) was added to complete media as a chemo-attractant.

    Techniques: Staining, In Vivo, Ex Vivo, Mouse Assay

    Expression and clinical relevance of IL-8 cytokine expression ( a ) Relative expression levels of IL-8 using RNA-Seq data from the TCGA data sets. The numbers above each tumour type (top of panel) represent the sample size used to calculate expression levels. LUAD, lung adenocarcinoma; LUSC, lung squamous carcinoma. Box plot represents first (lower bound) and third (upper bound) quartiles, whiskers represent 1.5 times the interquartile range. ( b ) Relative expression levels of IL-8 from the cancer cell line encyclopedia (CCLE) data set partitioned by designated tumour type. Analysis of variance test was used to test for statistical significance. NSCLC, non-small cell lung cancer. (c) Kaplan–Meier plots for overall survival based on IL-8 expression were generated for patients with basal-like ( n = 185), luminal A ( n = 459) and luminal B ( n = 308) breast cancers. ( d ) Kaplan–Meier plots for overall survival based on IL-8 expression for patients with NSCLC ( n = 1,404), lung adenocarcinoma ( n = 486) and LUSC ( n = 421). For c and d , above (high, in red) and below (low, in black) the median expression level was used as the threshold in all cohorts. ( e ) Kaplan–Meier plots for overall survival based on IL-8 expression for patients with renal adenocarcinomas ( n = 469) from the CGA data set. Above (high, in red) and below (low, in blue) the median expression level was used as the threshold. ( f ) Kaplan–Meier plot for overall survival based on IL-8 protein expression for patients with ovarian cancer ( n = 150). Scale bars, 200 μm.

    Journal: Nature communications

    Article Title: Tumour angiogenesis regulation by the miR-200 family

    doi: 10.1038/ncomms3427

    Figure Lengend Snippet: Expression and clinical relevance of IL-8 cytokine expression ( a ) Relative expression levels of IL-8 using RNA-Seq data from the TCGA data sets. The numbers above each tumour type (top of panel) represent the sample size used to calculate expression levels. LUAD, lung adenocarcinoma; LUSC, lung squamous carcinoma. Box plot represents first (lower bound) and third (upper bound) quartiles, whiskers represent 1.5 times the interquartile range. ( b ) Relative expression levels of IL-8 from the cancer cell line encyclopedia (CCLE) data set partitioned by designated tumour type. Analysis of variance test was used to test for statistical significance. NSCLC, non-small cell lung cancer. (c) Kaplan–Meier plots for overall survival based on IL-8 expression were generated for patients with basal-like ( n = 185), luminal A ( n = 459) and luminal B ( n = 308) breast cancers. ( d ) Kaplan–Meier plots for overall survival based on IL-8 expression for patients with NSCLC ( n = 1,404), lung adenocarcinoma ( n = 486) and LUSC ( n = 421). For c and d , above (high, in red) and below (low, in black) the median expression level was used as the threshold in all cohorts. ( e ) Kaplan–Meier plots for overall survival based on IL-8 expression for patients with renal adenocarcinomas ( n = 469) from the CGA data set. Above (high, in red) and below (low, in blue) the median expression level was used as the threshold. ( f ) Kaplan–Meier plot for overall survival based on IL-8 protein expression for patients with ovarian cancer ( n = 150). Scale bars, 200 μm.

    Article Snippet: For rescue experiments (RF24 cells), 10 ng ml−1 of recombinant human IL-8 (R & D Systems, Minneapolis, MN) was added to complete media as a chemo-attractant.

    Techniques: Expressing, RNA Sequencing Assay, Generated

    Direct and indirect anti-angiogenic effects by miR-200 ( a ) Expression levels of IL-8 and CXCL1 for ES2 and HeyA8 clones, n = 3. Data are averages ± s.e.m. ( b ) Expression levels of CXCL1 from murine lung cancer (344SQ) clones, n = 3. Data are averages ± s.e.m. ( c ) Expression levels of IL-8, CXCL1 and Ets-1 following transfection of RF24 cells. Data are averages ± s.e.m., n = 3. ( d ) ELISA for IL-8 (ES2 and HeyA8) and ( e ) CXCL1 (ES2) following transfection. Data are averages ± s.e.m., n = 3. ( f ) ELISA for IL-8 and CXCL1 supernatants from HeyA8 clones. Data are averages ± s.e.m., n = 3. ( g ) Relative expression levels of miR-200a, miR-200b and miR-210 in clinically isolated endothelial cells from normal ovary ( n = 2) versus carcinoma ( n = 6). Further comparison with normal ovarian epithelium ( n = 2) is shown. Data are averages ± s.e.m. P -values obtained with Mann–Whitney's t -test. ( h ) Relative luciferase activity normalized to empty control for the IL-8 3′-untranslated region (UTR) (wild-type, left; mutated, right). Data are averages ± s.e.m. P -values obtained with Student's t -test, n = 6. ( i ) Relative luciferase activity normalized to empty control for the CXCL1 3′-UTR (wild-type, left; mutated, right). Data are averages ± s.e.m. P -values obtained with Student's t -test, n = 6. ( j ) Direct effects on migration, n = 4 and ( k ) tube formation, n = 6 following RF24 transfection. Data are averages ± s.e.m. P -values were with Student's t -test. ( l ) Effects of exogenous recombinant IL-8 (10 ng ml −1 ) on migration and ( m ) tube formation following RF24 transfection, n = 6. Data are averages ± s.e.m. P -values were with Student's t -test. ( n ) Indirect effects on RF24 tube formation following incubation with HeyA8 supernatants, n = 6. Data are averages ± s.e.m. P -values were obtained with Student's t -test. Scale bar, 500 μm. ( o ) Representative images (left) and haemoglobin quantification (right) of an in-vivo matrigel plug assay, n = 2. White arrows demonstrate new blood vessel formation. Data are averages ± s.e.m. P -values obtained with Student's t -test. * P

    Journal: Nature communications

    Article Title: Tumour angiogenesis regulation by the miR-200 family

    doi: 10.1038/ncomms3427

    Figure Lengend Snippet: Direct and indirect anti-angiogenic effects by miR-200 ( a ) Expression levels of IL-8 and CXCL1 for ES2 and HeyA8 clones, n = 3. Data are averages ± s.e.m. ( b ) Expression levels of CXCL1 from murine lung cancer (344SQ) clones, n = 3. Data are averages ± s.e.m. ( c ) Expression levels of IL-8, CXCL1 and Ets-1 following transfection of RF24 cells. Data are averages ± s.e.m., n = 3. ( d ) ELISA for IL-8 (ES2 and HeyA8) and ( e ) CXCL1 (ES2) following transfection. Data are averages ± s.e.m., n = 3. ( f ) ELISA for IL-8 and CXCL1 supernatants from HeyA8 clones. Data are averages ± s.e.m., n = 3. ( g ) Relative expression levels of miR-200a, miR-200b and miR-210 in clinically isolated endothelial cells from normal ovary ( n = 2) versus carcinoma ( n = 6). Further comparison with normal ovarian epithelium ( n = 2) is shown. Data are averages ± s.e.m. P -values obtained with Mann–Whitney's t -test. ( h ) Relative luciferase activity normalized to empty control for the IL-8 3′-untranslated region (UTR) (wild-type, left; mutated, right). Data are averages ± s.e.m. P -values obtained with Student's t -test, n = 6. ( i ) Relative luciferase activity normalized to empty control for the CXCL1 3′-UTR (wild-type, left; mutated, right). Data are averages ± s.e.m. P -values obtained with Student's t -test, n = 6. ( j ) Direct effects on migration, n = 4 and ( k ) tube formation, n = 6 following RF24 transfection. Data are averages ± s.e.m. P -values were with Student's t -test. ( l ) Effects of exogenous recombinant IL-8 (10 ng ml −1 ) on migration and ( m ) tube formation following RF24 transfection, n = 6. Data are averages ± s.e.m. P -values were with Student's t -test. ( n ) Indirect effects on RF24 tube formation following incubation with HeyA8 supernatants, n = 6. Data are averages ± s.e.m. P -values were obtained with Student's t -test. Scale bar, 500 μm. ( o ) Representative images (left) and haemoglobin quantification (right) of an in-vivo matrigel plug assay, n = 2. White arrows demonstrate new blood vessel formation. Data are averages ± s.e.m. P -values obtained with Student's t -test. * P

    Article Snippet: For rescue experiments (RF24 cells), 10 ng ml−1 of recombinant human IL-8 (R & D Systems, Minneapolis, MN) was added to complete media as a chemo-attractant.

    Techniques: Expressing, Clone Assay, Transfection, Enzyme-linked Immunosorbent Assay, Isolation, MANN-WHITNEY, Luciferase, Activity Assay, Migration, Recombinant, Incubation, In Vivo, Matrigel Assay

    Migration of pericyte-like cells using conditioned media of miR-200-transfected cells ( a ) Migration of 10T1/2 pericyte-like cells at 6 h using conditioned media from HeyA8 (left) and ES2 (right) cells following miRNA transfection as a chemo-attractant. Data are averages ± s.e.m., n = 2. P -values obtained with Student's t -test. ( b ) Migration of 10T1/2 cells using complete media or complete media plus 10 ng ml −1 of IL-8 as a chemo-attractant. Data are averages ± s.e.m., n = 2. P -values obtained with Student's t -test. ( c ) Representative images of migrated pericyte-like cells using conditioned media from HeyA8 cells. ( d ) A model for miR-200 targeting of direct and indirect effects on angiogenesis. * P

    Journal: Nature communications

    Article Title: Tumour angiogenesis regulation by the miR-200 family

    doi: 10.1038/ncomms3427

    Figure Lengend Snippet: Migration of pericyte-like cells using conditioned media of miR-200-transfected cells ( a ) Migration of 10T1/2 pericyte-like cells at 6 h using conditioned media from HeyA8 (left) and ES2 (right) cells following miRNA transfection as a chemo-attractant. Data are averages ± s.e.m., n = 2. P -values obtained with Student's t -test. ( b ) Migration of 10T1/2 cells using complete media or complete media plus 10 ng ml −1 of IL-8 as a chemo-attractant. Data are averages ± s.e.m., n = 2. P -values obtained with Student's t -test. ( c ) Representative images of migrated pericyte-like cells using conditioned media from HeyA8 cells. ( d ) A model for miR-200 targeting of direct and indirect effects on angiogenesis. * P

    Article Snippet: For rescue experiments (RF24 cells), 10 ng ml−1 of recombinant human IL-8 (R & D Systems, Minneapolis, MN) was added to complete media as a chemo-attractant.

    Techniques: Migration, Transfection

    A high expression of CXCL8 in human ESCC tissues is associated with the poor prognosis of ESCC patients (A) A double immunofluorescence analysis was performed using anti-CXCL8 (green) and anti-CD11b (red) antibodies on human ESCC tissue. Nuclei were stained with DAPI (blue). Not only CD11b-positive macrophages but also the cancer nest was stained by CXCL8 in human ESCC tissue. Scale bar, 100 μm. (B) Immunohistochemical staining for CXCL8 in human ESCC tissue. Typical images of CXCL8 are shown: negative ( left ), low-intensity ( center ) and high-intensity ( right) compared to the corresponding normal esophageal epithelia. Scale bar, 100 μm. (C) Immunohistochemical staining for CXCR1 in human ESCC tissue. Typical images are shown: low-intensity ( left ) and high-intensity ( right ) compared to the corresponding normal esophageal epithelia. Scale bar, 100 μm. (D) Immunohistochemical staining for CXCR2 in human ESCC tissue. Typical images are shown: low-intensity ( left ) and high-intensity ( right ) compared to the corresponding normal esophageal epithelia. Scale bar, 100 μm. (E) Kaplan-Meier analysis of ESCC patients divided into three groups according to their expression of CXCL8: CXCL8-Negative group (n = 14), CXCL8-Low group (n = 43) and CXCL8-High group (n = 13). The log-rank test was performed to determine significance. * p

    Journal: Oncotarget

    Article Title: CXCL8 derived from tumor-associated macrophages and esophageal squamous cell carcinomas contributes to tumor progression by promoting migration and invasion of cancer cells

    doi: 10.18632/oncotarget.22526

    Figure Lengend Snippet: A high expression of CXCL8 in human ESCC tissues is associated with the poor prognosis of ESCC patients (A) A double immunofluorescence analysis was performed using anti-CXCL8 (green) and anti-CD11b (red) antibodies on human ESCC tissue. Nuclei were stained with DAPI (blue). Not only CD11b-positive macrophages but also the cancer nest was stained by CXCL8 in human ESCC tissue. Scale bar, 100 μm. (B) Immunohistochemical staining for CXCL8 in human ESCC tissue. Typical images of CXCL8 are shown: negative ( left ), low-intensity ( center ) and high-intensity ( right) compared to the corresponding normal esophageal epithelia. Scale bar, 100 μm. (C) Immunohistochemical staining for CXCR1 in human ESCC tissue. Typical images are shown: low-intensity ( left ) and high-intensity ( right ) compared to the corresponding normal esophageal epithelia. Scale bar, 100 μm. (D) Immunohistochemical staining for CXCR2 in human ESCC tissue. Typical images are shown: low-intensity ( left ) and high-intensity ( right ) compared to the corresponding normal esophageal epithelia. Scale bar, 100 μm. (E) Kaplan-Meier analysis of ESCC patients divided into three groups according to their expression of CXCL8: CXCL8-Negative group (n = 14), CXCL8-Low group (n = 43) and CXCL8-High group (n = 13). The log-rank test was performed to determine significance. * p

    Article Snippet: The cells were then treated with 0, 1, 10 or 100 ng/ml recombinant human CXCL8 (rhIL-8; R & D Systems).

    Techniques: Expressing, Immunofluorescence, Staining, Immunohistochemistry

    Akt and Erk1/2 were phosphorylated by CXCL8 through CXCR1 and CXCR2 in the ESCC cell lines (A) The mRNA levels of CXCR1 and CXCR2 in the ESCC cell lines were quantified by RT-PCR. (B) The protein level of CXCR1 and CXCR2 in the ESCC cell lines was confirmed by western blotting. Anti-CXCR1, CXCR2 and β-actin antibodies were used. (C) TE-8, TE-9 and TE-15 cells in serum-free conditions were treated with 10 ng/ml rhCXCL8 for 0, 10, 30 and 60 min. Western blotting was conducted with total protein extracted from ESCC cell lines using specific antibodies against Akt, p-Akt (Ser473), p-Akt (Thr308), Erk1/2, p-Erk1/2 (Thr202/Tyr204) and β-actin. Densitometric analysis of bands was performed with ImageJ (National Institutes of Health, Maryland, USA). The results are mean ± SEM. * p

    Journal: Oncotarget

    Article Title: CXCL8 derived from tumor-associated macrophages and esophageal squamous cell carcinomas contributes to tumor progression by promoting migration and invasion of cancer cells

    doi: 10.18632/oncotarget.22526

    Figure Lengend Snippet: Akt and Erk1/2 were phosphorylated by CXCL8 through CXCR1 and CXCR2 in the ESCC cell lines (A) The mRNA levels of CXCR1 and CXCR2 in the ESCC cell lines were quantified by RT-PCR. (B) The protein level of CXCR1 and CXCR2 in the ESCC cell lines was confirmed by western blotting. Anti-CXCR1, CXCR2 and β-actin antibodies were used. (C) TE-8, TE-9 and TE-15 cells in serum-free conditions were treated with 10 ng/ml rhCXCL8 for 0, 10, 30 and 60 min. Western blotting was conducted with total protein extracted from ESCC cell lines using specific antibodies against Akt, p-Akt (Ser473), p-Akt (Thr308), Erk1/2, p-Erk1/2 (Thr202/Tyr204) and β-actin. Densitometric analysis of bands was performed with ImageJ (National Institutes of Health, Maryland, USA). The results are mean ± SEM. * p

    Article Snippet: The cells were then treated with 0, 1, 10 or 100 ng/ml recombinant human CXCL8 (rhIL-8; R & D Systems).

    Techniques: Reverse Transcription Polymerase Chain Reaction, Western Blot

    CXCL8 promoted the migration and invasion of the TE-8 cells (A) (i) For the transwell migration assay, TE-8 cells were plated on the transwell in serum-free RPMI-1640 at 5.0 × 10 5 cells/well. rhCXCL8 was added in the upper chamber at 10 ng/ml. The cell inserts were set on 24-well plates in RPMI-1640 with 1% FBS for 24 h. The migrated cells on the underside of the membrane were stained by Diff-Quik and counted. The results are mean ± SEM. Scale bar, 100 μm. (ii) For the transwell invasion assay, TE-8 cells were seeded on a transwell coated with matrigel in serum-free RPMI-1640 at 5.0 × 10 5 cells/well. Recombinant human CXCL8 was added in the transwell at 100 ng/ml. The cell inserts were set on 24-well plates in RPMI-1640 with 1% FBS for 48 h. The invaded cells on the underside of the membrane were stained by Diff-Quik and counted. (B) (i) TE-8 cells were plated on the upper chamber with or without rhCXCL8 at 10 ng/ml combined with the inhibitor against PI3K (LY294002, 10 μM) or MEK1/2 (PD98059, 10 μM). DMSO (0.2 μl/ml) was added to negative control. After 24 h, the migrated cells were counted. (ii) TE-8 cells were plated on the upper transwell coated with matrigel. rhCXCL8 was added in the transwell combined with LY294002 (10 μM) or PD98059 (10 μM). DMSO (0.2 μl/ml) was added to negative control. After 48 h, the invaded cells were counted. (C) TE-8 cells were transfected with 20 nM siRNA targeting CXCR1 and CXCR2 . siNC was transfected to TE-8 as negative control. Effective knockdown of CXCR1 and CXCR2 was confirmed by western blotting using antibodies against CXCR1 and CXCR2. (D) (i) CXCR1 - or CXCR2- silenced TE-8 cells were plated on the upper transwell with rhCXCL8 at 10 ng/ml. After 24 h, the migrated cells were counted. (ii) CXCR1 - or CXCR2 -silenced TE-8 cells were plated on the upper transwell coated with matrigel. rhCXCL8 was added in the transwell at 100 ng/ml. After 48 h, the invaded cells were counted. (E) (i) TE-8 cells were plated on the upper transwell with rhCXCL8 at 10 ng/ml combined with the neutralizing antibody against CXCR1 (0.2 μg/ml) or CXCR2 (1.0 μg/ml). Mouse IgG was added to negative control. The concentrations of neutralizing antibodies were based on the manufacturer's instructions. (ii) TE-8 cells were plated on the upper transwell coated with matrigel. rhCXCL8 was added in the upper transwell at 100 ng/ml combined with the neutralizing antibody against CXCR1 (0.2 μg/ml) or CXCR2 (1.0 μg/ml). Mouse IgG was added to negative control. After 48 h, the invaded cells were counted. NS, not significant; * p

    Journal: Oncotarget

    Article Title: CXCL8 derived from tumor-associated macrophages and esophageal squamous cell carcinomas contributes to tumor progression by promoting migration and invasion of cancer cells

    doi: 10.18632/oncotarget.22526

    Figure Lengend Snippet: CXCL8 promoted the migration and invasion of the TE-8 cells (A) (i) For the transwell migration assay, TE-8 cells were plated on the transwell in serum-free RPMI-1640 at 5.0 × 10 5 cells/well. rhCXCL8 was added in the upper chamber at 10 ng/ml. The cell inserts were set on 24-well plates in RPMI-1640 with 1% FBS for 24 h. The migrated cells on the underside of the membrane were stained by Diff-Quik and counted. The results are mean ± SEM. Scale bar, 100 μm. (ii) For the transwell invasion assay, TE-8 cells were seeded on a transwell coated with matrigel in serum-free RPMI-1640 at 5.0 × 10 5 cells/well. Recombinant human CXCL8 was added in the transwell at 100 ng/ml. The cell inserts were set on 24-well plates in RPMI-1640 with 1% FBS for 48 h. The invaded cells on the underside of the membrane were stained by Diff-Quik and counted. (B) (i) TE-8 cells were plated on the upper chamber with or without rhCXCL8 at 10 ng/ml combined with the inhibitor against PI3K (LY294002, 10 μM) or MEK1/2 (PD98059, 10 μM). DMSO (0.2 μl/ml) was added to negative control. After 24 h, the migrated cells were counted. (ii) TE-8 cells were plated on the upper transwell coated with matrigel. rhCXCL8 was added in the transwell combined with LY294002 (10 μM) or PD98059 (10 μM). DMSO (0.2 μl/ml) was added to negative control. After 48 h, the invaded cells were counted. (C) TE-8 cells were transfected with 20 nM siRNA targeting CXCR1 and CXCR2 . siNC was transfected to TE-8 as negative control. Effective knockdown of CXCR1 and CXCR2 was confirmed by western blotting using antibodies against CXCR1 and CXCR2. (D) (i) CXCR1 - or CXCR2- silenced TE-8 cells were plated on the upper transwell with rhCXCL8 at 10 ng/ml. After 24 h, the migrated cells were counted. (ii) CXCR1 - or CXCR2 -silenced TE-8 cells were plated on the upper transwell coated with matrigel. rhCXCL8 was added in the transwell at 100 ng/ml. After 48 h, the invaded cells were counted. (E) (i) TE-8 cells were plated on the upper transwell with rhCXCL8 at 10 ng/ml combined with the neutralizing antibody against CXCR1 (0.2 μg/ml) or CXCR2 (1.0 μg/ml). Mouse IgG was added to negative control. The concentrations of neutralizing antibodies were based on the manufacturer's instructions. (ii) TE-8 cells were plated on the upper transwell coated with matrigel. rhCXCL8 was added in the upper transwell at 100 ng/ml combined with the neutralizing antibody against CXCR1 (0.2 μg/ml) or CXCR2 (1.0 μg/ml). Mouse IgG was added to negative control. After 48 h, the invaded cells were counted. NS, not significant; * p

    Article Snippet: The cells were then treated with 0, 1, 10 or 100 ng/ml recombinant human CXCL8 (rhIL-8; R & D Systems).

    Techniques: Migration, Transwell Migration Assay, Staining, Diff-Quik, Transwell Invasion Assay, Recombinant, Negative Control, Transfection, Western Blot

    TAM-like PBMo-derived macrophages induced the migration and invasion of TE-8 cells by activating PI3K and MEK1/2 signals through CXCL8 and CXCR1/2 interaction (A) PBMos (1.0 × 10 5 cells/well) were seeded on the lower chamber of 24-well plates with M-CSF (25 ng/ml) for 6 days to induce PBMo-derived macrophages, then incubated with 50% TE-8 CM to induce TAM-like PBMo-derived macrophages. After 2 days, the media were replaced with serum-free media. (i) TE-8 cells were plated on the upper transwell at 5.0 × 10 5 cells/well and set on the plate. After 24 h, the migrated cells were counted. (ii) TE-8 cells were plated on the upper transwell coated with matrigel at 5.0 × 10 5 cells/well and set on the plate. After 48 h, the invaded cells were counted. Scale bar, 100 μm. (B) (i) TE-8 cells were plated on the upper transwell with the inhibitor against PI3K (LY294002, 10 nM) or MEK1/2 (PD98059, 10 nM). DMSO (0.2 μl/ml) was added to negative control. After 24 h, the migrated cells were counted. (ii) TE-8 cells were plated on the upper matrigel-coated transwell with LY294002 (10 nM) or PD98059 (10 nM). DMSO (0.2 μl/ml) was added to negative control. After 48 h, the invaded cells were counted. (C) (i) TE-8 cells were plated on the upper transwell with the neutralizing antibodies against CXCR1 (0.2 μg/ml), CXCR2 (1.0 μg/ml) or CXCL8 (1.0 μg/ml). Mouse IgG was added to negative control. After 24 h, the migrated cells were counted. The concentrations of neutralizing antibody were based on manufacturer's instructions. (ii) TE-8 cells were plated on the upper matrigel-coated transwell with the neutralizing antibodies against CXCR1 (0.2 μg/ml), CXCR2 (1.0 μg/ml) or CXCL8 (1.0 μg/ml). Mouse IgG was added to negative control. After 48 h, the invaded cells were counted. The results are mean ± SEM. * p

    Journal: Oncotarget

    Article Title: CXCL8 derived from tumor-associated macrophages and esophageal squamous cell carcinomas contributes to tumor progression by promoting migration and invasion of cancer cells

    doi: 10.18632/oncotarget.22526

    Figure Lengend Snippet: TAM-like PBMo-derived macrophages induced the migration and invasion of TE-8 cells by activating PI3K and MEK1/2 signals through CXCL8 and CXCR1/2 interaction (A) PBMos (1.0 × 10 5 cells/well) were seeded on the lower chamber of 24-well plates with M-CSF (25 ng/ml) for 6 days to induce PBMo-derived macrophages, then incubated with 50% TE-8 CM to induce TAM-like PBMo-derived macrophages. After 2 days, the media were replaced with serum-free media. (i) TE-8 cells were plated on the upper transwell at 5.0 × 10 5 cells/well and set on the plate. After 24 h, the migrated cells were counted. (ii) TE-8 cells were plated on the upper transwell coated with matrigel at 5.0 × 10 5 cells/well and set on the plate. After 48 h, the invaded cells were counted. Scale bar, 100 μm. (B) (i) TE-8 cells were plated on the upper transwell with the inhibitor against PI3K (LY294002, 10 nM) or MEK1/2 (PD98059, 10 nM). DMSO (0.2 μl/ml) was added to negative control. After 24 h, the migrated cells were counted. (ii) TE-8 cells were plated on the upper matrigel-coated transwell with LY294002 (10 nM) or PD98059 (10 nM). DMSO (0.2 μl/ml) was added to negative control. After 48 h, the invaded cells were counted. (C) (i) TE-8 cells were plated on the upper transwell with the neutralizing antibodies against CXCR1 (0.2 μg/ml), CXCR2 (1.0 μg/ml) or CXCL8 (1.0 μg/ml). Mouse IgG was added to negative control. After 24 h, the migrated cells were counted. The concentrations of neutralizing antibody were based on manufacturer's instructions. (ii) TE-8 cells were plated on the upper matrigel-coated transwell with the neutralizing antibodies against CXCR1 (0.2 μg/ml), CXCR2 (1.0 μg/ml) or CXCL8 (1.0 μg/ml). Mouse IgG was added to negative control. After 48 h, the invaded cells were counted. The results are mean ± SEM. * p

    Article Snippet: The cells were then treated with 0, 1, 10 or 100 ng/ml recombinant human CXCL8 (rhIL-8; R & D Systems).

    Techniques: Derivative Assay, Migration, Incubation, Negative Control

    A proposed model of the interaction between ESCC cells and TAMs through CXCL8 in tumor microenvironment Many humoral factors, including IL-4, induced macrophages into TAMs. Then, expression level of CXCL8 was up-regulated in TAMs. CXCL8 secreted from TAMs, as well as from ESCC cells, contributed ESCC cells' migration and invasion by activating Akt and Erk1/2 signals through CXCR1/CXCR2 of ESCC cells.

    Journal: Oncotarget

    Article Title: CXCL8 derived from tumor-associated macrophages and esophageal squamous cell carcinomas contributes to tumor progression by promoting migration and invasion of cancer cells

    doi: 10.18632/oncotarget.22526

    Figure Lengend Snippet: A proposed model of the interaction between ESCC cells and TAMs through CXCL8 in tumor microenvironment Many humoral factors, including IL-4, induced macrophages into TAMs. Then, expression level of CXCL8 was up-regulated in TAMs. CXCL8 secreted from TAMs, as well as from ESCC cells, contributed ESCC cells' migration and invasion by activating Akt and Erk1/2 signals through CXCR1/CXCR2 of ESCC cells.

    Article Snippet: The cells were then treated with 0, 1, 10 or 100 ng/ml recombinant human CXCL8 (rhIL-8; R & D Systems).

    Techniques: Expressing, Migration

    Induction of CXCL8 in PBMo-derived macrophages stimulated with TECM (A) The mRNA level of CXCL8 in PBMo-derived macrophages stimulated with 50% TECM or 50% Het-1A CM was determined by quantitative RT-PCR. The data were normalized to GAPDH as an internal control. Data are mean ± SEM in triplicate. ** p

    Journal: Oncotarget

    Article Title: CXCL8 derived from tumor-associated macrophages and esophageal squamous cell carcinomas contributes to tumor progression by promoting migration and invasion of cancer cells

    doi: 10.18632/oncotarget.22526

    Figure Lengend Snippet: Induction of CXCL8 in PBMo-derived macrophages stimulated with TECM (A) The mRNA level of CXCL8 in PBMo-derived macrophages stimulated with 50% TECM or 50% Het-1A CM was determined by quantitative RT-PCR. The data were normalized to GAPDH as an internal control. Data are mean ± SEM in triplicate. ** p

    Article Snippet: The cells were then treated with 0, 1, 10 or 100 ng/ml recombinant human CXCL8 (rhIL-8; R & D Systems).

    Techniques: Derivative Assay, Quantitative RT-PCR

    Immunohistochemical staining of CXCL8 in cervical cancer tissues ( A ) Negative expression of CXCL8 in normal cervical tissue; ( B ) weak expression of CXCL8 in normal cervical tissue; ( C ) moderate expression of CXCL8 in normal cervical tissue; ( D ) strong expression of CXCL8 in normal cervical tissue; ( E ) negative expression of CXCL8 in cervical squamous cell carcinoma tissue; ( F ) weak expression of CXCL8 in cervical squamous cell carcinoma tissue; ( G ) moderate expression of CXCL8 in cervical squamous cell carcinoma tissue; ( H ) strong expression of CXCL8 in cervical squamous cell carcinoma tissue; ( I ) negative expression of CXCL8 in cervical adenocarcinoma tissue; ( J ) weak expression of CXCL8 in cervical adenocarcinoma tissue; ( K ) moderate expression of CXCL8 in cervical adenocarcinoma tissue; ( L ) strong expression of CXCL8 in cervical adenocarcinoma tissue.

    Journal: Bioscience Reports

    Article Title: The clinical and prognostic value of CXCL8 in cervical carcinoma patients: immunohistochemical analysis

    doi: 10.1042/BSR20171021

    Figure Lengend Snippet: Immunohistochemical staining of CXCL8 in cervical cancer tissues ( A ) Negative expression of CXCL8 in normal cervical tissue; ( B ) weak expression of CXCL8 in normal cervical tissue; ( C ) moderate expression of CXCL8 in normal cervical tissue; ( D ) strong expression of CXCL8 in normal cervical tissue; ( E ) negative expression of CXCL8 in cervical squamous cell carcinoma tissue; ( F ) weak expression of CXCL8 in cervical squamous cell carcinoma tissue; ( G ) moderate expression of CXCL8 in cervical squamous cell carcinoma tissue; ( H ) strong expression of CXCL8 in cervical squamous cell carcinoma tissue; ( I ) negative expression of CXCL8 in cervical adenocarcinoma tissue; ( J ) weak expression of CXCL8 in cervical adenocarcinoma tissue; ( K ) moderate expression of CXCL8 in cervical adenocarcinoma tissue; ( L ) strong expression of CXCL8 in cervical adenocarcinoma tissue.

    Article Snippet: Low expression of CXCL8 was defined as 0–4 score; high expression of CXCL8 was defined as more than 4 score.

    Techniques: Immunohistochemistry, Staining, Expressing

    CXCL8 mRNA and protein expressions in cervical cancer tissues and cell lines ( A ) CXCL8 high expression is observed in cervical cancer tissues compared with normal cervical tissues (GSE9750). ( B ) CXCL8 is overexpressed in cervical cancer tissues compared with normal cervical tissues and cervical intraepithelial lesions’ tissues (GSE7803). ( C ) CXCL8 mRNA expression is higher in cervical cancer tissues than adjacent normal cervical tissues. ( D ) CXCL8 protein expression is higher in cervical cancer tissues than adjacent normal cervical tissues. ( E ) Levels of CXCL8 mRNA are increased in cervical cancer cell lines compared with normal cervical epithelial cells lines. ( F ) CXCL8 protein expression is highly expressed in cervical cancer cell lines compared with normal cervical epithelial cell lines.

    Journal: Bioscience Reports

    Article Title: The clinical and prognostic value of CXCL8 in cervical carcinoma patients: immunohistochemical analysis

    doi: 10.1042/BSR20171021

    Figure Lengend Snippet: CXCL8 mRNA and protein expressions in cervical cancer tissues and cell lines ( A ) CXCL8 high expression is observed in cervical cancer tissues compared with normal cervical tissues (GSE9750). ( B ) CXCL8 is overexpressed in cervical cancer tissues compared with normal cervical tissues and cervical intraepithelial lesions’ tissues (GSE7803). ( C ) CXCL8 mRNA expression is higher in cervical cancer tissues than adjacent normal cervical tissues. ( D ) CXCL8 protein expression is higher in cervical cancer tissues than adjacent normal cervical tissues. ( E ) Levels of CXCL8 mRNA are increased in cervical cancer cell lines compared with normal cervical epithelial cells lines. ( F ) CXCL8 protein expression is highly expressed in cervical cancer cell lines compared with normal cervical epithelial cell lines.

    Article Snippet: Low expression of CXCL8 was defined as 0–4 score; high expression of CXCL8 was defined as more than 4 score.

    Techniques: Expressing

    The prognostic significance of CXCL8 in cervical cancer ( A ) CXCL8 high expression is negatively associated with overall survival in 108 cervical cancer patients from our study. ( B ) The Cancer Genome Atlas database including 246 cervical cancer patients shows that CXCL8 protein’s high expression had shorter overall survival compared in patients with CXCL8 protein’s low expression.

    Journal: Bioscience Reports

    Article Title: The clinical and prognostic value of CXCL8 in cervical carcinoma patients: immunohistochemical analysis

    doi: 10.1042/BSR20171021

    Figure Lengend Snippet: The prognostic significance of CXCL8 in cervical cancer ( A ) CXCL8 high expression is negatively associated with overall survival in 108 cervical cancer patients from our study. ( B ) The Cancer Genome Atlas database including 246 cervical cancer patients shows that CXCL8 protein’s high expression had shorter overall survival compared in patients with CXCL8 protein’s low expression.

    Article Snippet: Low expression of CXCL8 was defined as 0–4 score; high expression of CXCL8 was defined as more than 4 score.

    Techniques: Expressing

    CXCL8 + naïve T cells are enriched in CD31 + T cells. (A–D) Fresh CXCL8 + umbilical cord blood naïve T cells were analyzed for expression of surface and intracellular marks of activation and differentiation. Representative FACS plots and percentages are shown. Gated on CXCL8 + naïve T cells. (E) Graphical summary of CXCL8 expression in CD31 + and CD31 − T cells (E). n = 4–8, *, P

    Journal: Journal of immunology (Baltimore, Md. : 1950)

    Article Title: Human naïve T cells express functional CXCL8 and promote tumorigenesis

    doi: 10.4049/jimmunol.1700755

    Figure Lengend Snippet: CXCL8 + naïve T cells are enriched in CD31 + T cells. (A–D) Fresh CXCL8 + umbilical cord blood naïve T cells were analyzed for expression of surface and intracellular marks of activation and differentiation. Representative FACS plots and percentages are shown. Gated on CXCL8 + naïve T cells. (E) Graphical summary of CXCL8 expression in CD31 + and CD31 − T cells (E). n = 4–8, *, P

    Article Snippet: Antibodies for flow cytometry were CCR7, CD3, CD4, CD8, CD25, CD31, CD45RA, CD45RO, CD62L, CD69, CD154, CXCR2, CXCR3, CXCL8, IFN-γ, IL-4, IL-10, IL-17, and IL-22 (BD), CD57, CD122, Foxp3 (eBiosciences/ThermoFischer), KLRG-1 (BioLegend).

    Techniques: Expressing, Activation Assay, FACS

    Human naïve T cells spontaneously express CXCL8. (A) Gating strategy used to identify T cell subsets. Naïve T cells: CD3 + CD4 + CD45RA + CD45RO − CD62L + CCR7 + . Memory T cells: CD3 + CD4 + CD45RA − CD45RO + CD62L − CCR7 − . (B–E) Cytokine profile of cord blood and peripheral T cells. Intracellular cytokines were analyzed by FACS. Representative FACS plots and percentages are shown (B, C). (D–E) Graphical summary of CXCL8 (D) and IFN-γ (E) FACS values in human T cell subsets. n = 4–8, *, P

    Journal: Journal of immunology (Baltimore, Md. : 1950)

    Article Title: Human naïve T cells express functional CXCL8 and promote tumorigenesis

    doi: 10.4049/jimmunol.1700755

    Figure Lengend Snippet: Human naïve T cells spontaneously express CXCL8. (A) Gating strategy used to identify T cell subsets. Naïve T cells: CD3 + CD4 + CD45RA + CD45RO − CD62L + CCR7 + . Memory T cells: CD3 + CD4 + CD45RA − CD45RO + CD62L − CCR7 − . (B–E) Cytokine profile of cord blood and peripheral T cells. Intracellular cytokines were analyzed by FACS. Representative FACS plots and percentages are shown (B, C). (D–E) Graphical summary of CXCL8 (D) and IFN-γ (E) FACS values in human T cell subsets. n = 4–8, *, P

    Article Snippet: Antibodies for flow cytometry were CCR7, CD3, CD4, CD8, CD25, CD31, CD45RA, CD45RO, CD62L, CD69, CD154, CXCR2, CXCR3, CXCL8, IFN-γ, IL-4, IL-10, IL-17, and IL-22 (BD), CD57, CD122, Foxp3 (eBiosciences/ThermoFischer), KLRG-1 (BioLegend).

    Techniques: FACS

    Human naïve T cells retain CXCL8 expression in vitro under homeostatic and activation conditions. (A–D) Results are shown as the kinetic CXCL8 (A, B) and IFN-γ (C, D) expression in naïve T cells in cord blood (A, C) and peripheral blood (B, D) upon activation with αCD3/αCD28 beads. (E) Expression plots for CXCL8 and IFN-γ following in vitro activation. (F–G) Results are shown as the kinetic CXCL8 (F) and IFN-γ (G) expression in naïve T cells in the presence of IL-7 and IL-15 from human tonsil. Average of 3–4 experiments is shown.

    Journal: Journal of immunology (Baltimore, Md. : 1950)

    Article Title: Human naïve T cells express functional CXCL8 and promote tumorigenesis

    doi: 10.4049/jimmunol.1700755

    Figure Lengend Snippet: Human naïve T cells retain CXCL8 expression in vitro under homeostatic and activation conditions. (A–D) Results are shown as the kinetic CXCL8 (A, B) and IFN-γ (C, D) expression in naïve T cells in cord blood (A, C) and peripheral blood (B, D) upon activation with αCD3/αCD28 beads. (E) Expression plots for CXCL8 and IFN-γ following in vitro activation. (F–G) Results are shown as the kinetic CXCL8 (F) and IFN-γ (G) expression in naïve T cells in the presence of IL-7 and IL-15 from human tonsil. Average of 3–4 experiments is shown.

    Article Snippet: Antibodies for flow cytometry were CCR7, CD3, CD4, CD8, CD25, CD31, CD45RA, CD45RO, CD62L, CD69, CD154, CXCR2, CXCR3, CXCL8, IFN-γ, IL-4, IL-10, IL-17, and IL-22 (BD), CD57, CD122, Foxp3 (eBiosciences/ThermoFischer), KLRG-1 (BioLegend).

    Techniques: Expressing, In Vitro, Activation Assay

    Naive T cell-derived CXCL8 promotes tumor growth and neutrophil migration. (A–B) Human primary cells (OC8) were mixed with cord blood naïve T cells and subcutaneously injected into NSG mice. Mice received daily intraperitoneal PBS or Raparixin (5mg/kg) injection. Following two-weeks, tumors were recovered and weighted. Representative tumors are presented in (A) and graphical summary of tumor size (B). n = 5/group. *, P

    Journal: Journal of immunology (Baltimore, Md. : 1950)

    Article Title: Human naïve T cells express functional CXCL8 and promote tumorigenesis

    doi: 10.4049/jimmunol.1700755

    Figure Lengend Snippet: Naive T cell-derived CXCL8 promotes tumor growth and neutrophil migration. (A–B) Human primary cells (OC8) were mixed with cord blood naïve T cells and subcutaneously injected into NSG mice. Mice received daily intraperitoneal PBS or Raparixin (5mg/kg) injection. Following two-weeks, tumors were recovered and weighted. Representative tumors are presented in (A) and graphical summary of tumor size (B). n = 5/group. *, P

    Article Snippet: Antibodies for flow cytometry were CCR7, CD3, CD4, CD8, CD25, CD31, CD45RA, CD45RO, CD62L, CD69, CD154, CXCR2, CXCR3, CXCL8, IFN-γ, IL-4, IL-10, IL-17, and IL-22 (BD), CD57, CD122, Foxp3 (eBiosciences/ThermoFischer), KLRG-1 (BioLegend).

    Techniques: Derivative Assay, Migration, Injection, Mouse Assay

    Interactions among serum biomarkers. Plasma levels of IL-1 β , IL-6, CXCL-8, TNF- α , IL-12p70, IFN- γ , IL-2, IL-4, IL-5, and IL-10 were measured in the serum of healthy donors and P. vivax -infected subjects. Each connecting line represents a significant correlation between a pair of biomarkers. Dashed lines represent negative correlations. Solid lines represent positive correlations, and the degree of significance is represented by the line thickness. Filled circles represent higher producers and open circles represent low producers of a specific biomarker. (a) and (b) represent biomarker interactions in HD (a) and P. vivax -infected subjects (b). (c) and (d) represent biomarker interaction in P. vivax presenting low (low) and high parasitemia (right). (e) represents biomarkers interactions that are preserved upon P. vivax infection and interactions gained or lost during malaria. Circle layouts underscore each biomarker by globular nodes (° or •, as the comprise

    Journal: Journal of Immunology Research

    Article Title: The Robust and Modulated Biomarker Network Elicited by the Plasmodium vivax Infection Is Mainly Mediated by the IL-6/IL-10 Axis and Is Associated with the Parasite Load

    doi: 10.1155/2014/318250

    Figure Lengend Snippet: Interactions among serum biomarkers. Plasma levels of IL-1 β , IL-6, CXCL-8, TNF- α , IL-12p70, IFN- γ , IL-2, IL-4, IL-5, and IL-10 were measured in the serum of healthy donors and P. vivax -infected subjects. Each connecting line represents a significant correlation between a pair of biomarkers. Dashed lines represent negative correlations. Solid lines represent positive correlations, and the degree of significance is represented by the line thickness. Filled circles represent higher producers and open circles represent low producers of a specific biomarker. (a) and (b) represent biomarker interactions in HD (a) and P. vivax -infected subjects (b). (c) and (d) represent biomarker interaction in P. vivax presenting low (low) and high parasitemia (right). (e) represents biomarkers interactions that are preserved upon P. vivax infection and interactions gained or lost during malaria. Circle layouts underscore each biomarker by globular nodes (° or •, as the comprise

    Article Snippet: The serum biomarkers IL-1β , IL-5, CXCL-8, and IL-12p70 were measured by ELISA (Kit Human BD OptEIA Set II, BD Biosciences Pharmingen, EUA).

    Techniques: Infection, Biomarker Assay

    High levels of IL-6, CXCL-8, IFN- γ , IL-5, and IL-10 are found in the circulation of patients infected with P. vivax . The biomarkers IL-1 β , IL-6, CXCL-8, TNF- α , IL-12p70, IFN- γ , IL-2, IL-4, IL-5, and IL-10 were measured in the serum of healthy donors (open circles) and P. vivax -infected subjects (closed circles). Levels of biomarkers were measured employing ELISA or Cytometric Bead Array (CBA).

    Journal: Journal of Immunology Research

    Article Title: The Robust and Modulated Biomarker Network Elicited by the Plasmodium vivax Infection Is Mainly Mediated by the IL-6/IL-10 Axis and Is Associated with the Parasite Load

    doi: 10.1155/2014/318250

    Figure Lengend Snippet: High levels of IL-6, CXCL-8, IFN- γ , IL-5, and IL-10 are found in the circulation of patients infected with P. vivax . The biomarkers IL-1 β , IL-6, CXCL-8, TNF- α , IL-12p70, IFN- γ , IL-2, IL-4, IL-5, and IL-10 were measured in the serum of healthy donors (open circles) and P. vivax -infected subjects (closed circles). Levels of biomarkers were measured employing ELISA or Cytometric Bead Array (CBA).

    Article Snippet: The serum biomarkers IL-1β , IL-5, CXCL-8, and IL-12p70 were measured by ELISA (Kit Human BD OptEIA Set II, BD Biosciences Pharmingen, EUA).

    Techniques: Infection, Enzyme-linked Immunosorbent Assay, Crocin Bleaching Assay

    Tumor cell viability in human ovarian tumor cell suspensions following treatment with oncolytic adenovirus, recombinant IL-8 or anti-IL-8 antibody Cell viability was measured using a MTS assay on days 7-13 after the start of incubation. Viability is presented as percentage of mock group cell viability. Ad = Ad5/3-d24. aIL8=anti-IL-8=anti-IL-8 neutralizing antibody. rIL8=IL-8 = recombinant IL-8. Asterisks indicate the significance of findings: * (p

    Journal: Oncotarget

    Article Title: Interleukin 8 activity influences the efficacy of adenoviral oncolytic immunotherapy in cancer patients

    doi: 10.18632/oncotarget.23967

    Figure Lengend Snippet: Tumor cell viability in human ovarian tumor cell suspensions following treatment with oncolytic adenovirus, recombinant IL-8 or anti-IL-8 antibody Cell viability was measured using a MTS assay on days 7-13 after the start of incubation. Viability is presented as percentage of mock group cell viability. Ad = Ad5/3-d24. aIL8=anti-IL-8=anti-IL-8 neutralizing antibody. rIL8=IL-8 = recombinant IL-8. Asterisks indicate the significance of findings: * (p

    Article Snippet: In addition, recombinant human IL-8 (Peprotech, Rocky Hill, NJ) or neutralizing anti-IL-8 antibody was added to the co-culture at the final concentration of 500 ng/ml and 2 μg/ml, respectively.

    Techniques: Recombinant, MTS Assay, Incubation

    Effect of IL-8 change status on overall survival and treatment responses Asterisks indicate statistical significance: *** p

    Journal: Oncotarget

    Article Title: Interleukin 8 activity influences the efficacy of adenoviral oncolytic immunotherapy in cancer patients

    doi: 10.18632/oncotarget.23967

    Figure Lengend Snippet: Effect of IL-8 change status on overall survival and treatment responses Asterisks indicate statistical significance: *** p

    Article Snippet: In addition, recombinant human IL-8 (Peprotech, Rocky Hill, NJ) or neutralizing anti-IL-8 antibody was added to the co-culture at the final concentration of 500 ng/ml and 2 μg/ml, respectively.

    Techniques:

    Tumor IL-8 and IL-8 receptor mRNA expression (Panel A) mRNA expression for IL-8 and its receptors CXC chemokine receptor 1 (CXCR1) and 2 (CXCR2) was quantified from pre- and post-treatment tumor samples. Expression is presented as log2-transformed values. (Panel B) Overall survival in different IL-8 expression groups. Patients were grouped based on the pre-treatment IL-8 expression level. Median OS was 174 days in the low expression group and 72 days in the high expression group (n=15, p=0.058). (Panel C) IL-8 expression change was determined as the difference between pre- and post-treatment expression values. No significant difference in survival between decrease and increase groups was found.

    Journal: Oncotarget

    Article Title: Interleukin 8 activity influences the efficacy of adenoviral oncolytic immunotherapy in cancer patients

    doi: 10.18632/oncotarget.23967

    Figure Lengend Snippet: Tumor IL-8 and IL-8 receptor mRNA expression (Panel A) mRNA expression for IL-8 and its receptors CXC chemokine receptor 1 (CXCR1) and 2 (CXCR2) was quantified from pre- and post-treatment tumor samples. Expression is presented as log2-transformed values. (Panel B) Overall survival in different IL-8 expression groups. Patients were grouped based on the pre-treatment IL-8 expression level. Median OS was 174 days in the low expression group and 72 days in the high expression group (n=15, p=0.058). (Panel C) IL-8 expression change was determined as the difference between pre- and post-treatment expression values. No significant difference in survival between decrease and increase groups was found.

    Article Snippet: In addition, recombinant human IL-8 (Peprotech, Rocky Hill, NJ) or neutralizing anti-IL-8 antibody was added to the co-culture at the final concentration of 500 ng/ml and 2 μg/ml, respectively.

    Techniques: Expressing, Transformation Assay

    Overall survival in patients with different baseline T cell activity status, neutrophil-to-lymphocyte ratio (NLR) and IL-8 levels Patients with increase or decrease in post-treatment anti-survivin were included in the T cell activity group, while patients with no change in the ELISPOT were assigned to anergy group. Asterisks indicate statistical significance: *** p

    Journal: Oncotarget

    Article Title: Interleukin 8 activity influences the efficacy of adenoviral oncolytic immunotherapy in cancer patients

    doi: 10.18632/oncotarget.23967

    Figure Lengend Snippet: Overall survival in patients with different baseline T cell activity status, neutrophil-to-lymphocyte ratio (NLR) and IL-8 levels Patients with increase or decrease in post-treatment anti-survivin were included in the T cell activity group, while patients with no change in the ELISPOT were assigned to anergy group. Asterisks indicate statistical significance: *** p

    Article Snippet: In addition, recombinant human IL-8 (Peprotech, Rocky Hill, NJ) or neutralizing anti-IL-8 antibody was added to the co-culture at the final concentration of 500 ng/ml and 2 μg/ml, respectively.

    Techniques: Activity Assay, Enzyme-linked Immunospot

    Cytotoxic T cell activation and IL-8 production in ovarian tumor derived TIL and TAN co-cultures obtained from human patients (Panels A-B) . Activation marker CD25 was used for samples from Tumor 1 and activation marker CD69 for samples from Tumor 7. (Panels C-D) IL-8 concentration was measured after a 24 hour incubation from TIL-TAN co-cultures or TILs/TANs alone. Ad = Ad5/3-d24. rIL8 = recombinant IL-8. Asterisks indicate the significance of findings: * (p

    Journal: Oncotarget

    Article Title: Interleukin 8 activity influences the efficacy of adenoviral oncolytic immunotherapy in cancer patients

    doi: 10.18632/oncotarget.23967

    Figure Lengend Snippet: Cytotoxic T cell activation and IL-8 production in ovarian tumor derived TIL and TAN co-cultures obtained from human patients (Panels A-B) . Activation marker CD25 was used for samples from Tumor 1 and activation marker CD69 for samples from Tumor 7. (Panels C-D) IL-8 concentration was measured after a 24 hour incubation from TIL-TAN co-cultures or TILs/TANs alone. Ad = Ad5/3-d24. rIL8 = recombinant IL-8. Asterisks indicate the significance of findings: * (p

    Article Snippet: In addition, recombinant human IL-8 (Peprotech, Rocky Hill, NJ) or neutralizing anti-IL-8 antibody was added to the co-culture at the final concentration of 500 ng/ml and 2 μg/ml, respectively.

    Techniques: Activation Assay, Derivative Assay, Marker, Concentration Assay, Incubation, Recombinant

    Overall survival and treatment responses in patients with normal and high pre-treatment IL-8 levels in blood and IL-8 concentrations in different tumor groups Patients were grouped based on the laboratory reference value for normal IL-8 (

    Journal: Oncotarget

    Article Title: Interleukin 8 activity influences the efficacy of adenoviral oncolytic immunotherapy in cancer patients

    doi: 10.18632/oncotarget.23967

    Figure Lengend Snippet: Overall survival and treatment responses in patients with normal and high pre-treatment IL-8 levels in blood and IL-8 concentrations in different tumor groups Patients were grouped based on the laboratory reference value for normal IL-8 (

    Article Snippet: In addition, recombinant human IL-8 (Peprotech, Rocky Hill, NJ) or neutralizing anti-IL-8 antibody was added to the co-culture at the final concentration of 500 ng/ml and 2 μg/ml, respectively.

    Techniques:

    Role of the IL-8/IL-8R axis in the ECM invasion of tumor cells

    Journal: Cancer research

    Article Title: IL-8 signaling plays a critical role in the epithelial-mesenchymal transition of human carcinoma cells

    doi: 10.1158/0008-5472.CAN-11-0156

    Figure Lengend Snippet: Role of the IL-8/IL-8R axis in the ECM invasion of tumor cells

    Article Snippet: For treatment with IL-8, adherent tumor cells were cultured in the presence of recombinant IL-8 (Peprotech, Rocky Hill, NJ) at the indicated concentrations for 72 hours in serum-free medium.

    Techniques:

    IL-8 induces invasiveness of breast cancer cells in a Brachyury-dependent manner

    Journal: Cancer research

    Article Title: IL-8 signaling plays a critical role in the epithelial-mesenchymal transition of human carcinoma cells

    doi: 10.1158/0008-5472.CAN-11-0156

    Figure Lengend Snippet: IL-8 induces invasiveness of breast cancer cells in a Brachyury-dependent manner

    Article Snippet: For treatment with IL-8, adherent tumor cells were cultured in the presence of recombinant IL-8 (Peprotech, Rocky Hill, NJ) at the indicated concentrations for 72 hours in serum-free medium.

    Techniques:

    Brachyury induces IL-8 and IL-8R expression in epithelial tumor cells

    Journal: Cancer research

    Article Title: IL-8 signaling plays a critical role in the epithelial-mesenchymal transition of human carcinoma cells

    doi: 10.1158/0008-5472.CAN-11-0156

    Figure Lengend Snippet: Brachyury induces IL-8 and IL-8R expression in epithelial tumor cells

    Article Snippet: For treatment with IL-8, adherent tumor cells were cultured in the presence of recombinant IL-8 (Peprotech, Rocky Hill, NJ) at the indicated concentrations for 72 hours in serum-free medium.

    Techniques: Expressing

    Role of IL-8 signaling in EMT

    Journal: Cancer research

    Article Title: IL-8 signaling plays a critical role in the epithelial-mesenchymal transition of human carcinoma cells

    doi: 10.1158/0008-5472.CAN-11-0156

    Figure Lengend Snippet: Role of IL-8 signaling in EMT

    Article Snippet: For treatment with IL-8, adherent tumor cells were cultured in the presence of recombinant IL-8 (Peprotech, Rocky Hill, NJ) at the indicated concentrations for 72 hours in serum-free medium.

    Techniques:

    IL-8 induces mesenchymal characteristics in tumor cells

    Journal: Cancer research

    Article Title: IL-8 signaling plays a critical role in the epithelial-mesenchymal transition of human carcinoma cells

    doi: 10.1158/0008-5472.CAN-11-0156

    Figure Lengend Snippet: IL-8 induces mesenchymal characteristics in tumor cells

    Article Snippet: For treatment with IL-8, adherent tumor cells were cultured in the presence of recombinant IL-8 (Peprotech, Rocky Hill, NJ) at the indicated concentrations for 72 hours in serum-free medium.

    Techniques:

    PM2.5 exposure induced upregulation of VEGFA production in human bronchial epithelial cells. (A) Beas-2B cells were left untreated or were treated with different concentrations of PM2.5 (12.5, 25, 50, and 100 μg/mL) for 24 h. The production of VEGFA, IL1B, IL6, CXCL8 and TNF was then detected in the cell culture supernatant using ELISA (*, P

    Journal: Autophagy

    Article Title: TP53-dependent autophagy links the ATR-CHEK1 axis activation to proinflammatory VEGFA production in human bronchial epithelial cells exposed to fine particulate matter (PM2.5)

    doi: 10.1080/15548627.2016.1204496

    Figure Lengend Snippet: PM2.5 exposure induced upregulation of VEGFA production in human bronchial epithelial cells. (A) Beas-2B cells were left untreated or were treated with different concentrations of PM2.5 (12.5, 25, 50, and 100 μg/mL) for 24 h. The production of VEGFA, IL1B, IL6, CXCL8 and TNF was then detected in the cell culture supernatant using ELISA (*, P

    Article Snippet: Cytokine production in the cell culture supernatants was quantified with human VEGFA (eBioscience, BMS277/2), IL1B (eBioscience, BMS224/2), IL6 (eBioscience, BMS213HS), CXCL8 (eBioscience, BMS204/3) and TNF (eBioscience, BMS223INST) immunoassay kits.

    Techniques: Cell Culture, Enzyme-linked Immunosorbent Assay

    Staining human neutrophils by anti-IL-17A (AF-317-NA) polyclonal antibodies (Abs). (A) Immunofluorescence (top panels) and immunohistochemistry (lower panels) stainings of two FFPE cases of human pustular psoriasis using anti-IL-17A (AF-317-NA) and anti-CD66b Abs (as labeled). Top panels show DAPI, FITC channel, and merge to recognize neutrophil shape; lower panels show different magnification of IHC and double IHC to characterize IL-17A + cells with the neutrophil marker CD66b. (B) Cytospins of neutrophils, either untreated (top panels) or treated with 5 µM R848 (bottom panels) for 3 h, were stained with anti-IL-17A (AF-317-NA, left panels) and anti-CXCL8 (right panels) Abs. Original magnification 200× [first row in (A) and left image in third row, scale bar 100 µm] and 400× [second row in (A) , center/right images in third row in (A) , as well as in (B) , scale bar 50 µm]. Images of the second row in (A) represent magnifications of images in first row. (C) AF-317-NA immunoblot of lysates from neutrophils either freshly isolated (T 0 , from two donors) or incubated for 3 h with or without 2 µg/ml IL-6 plus 0.2 µg/ml IL-23 (low), 20 µg/ml IL-6 plus 2 µg/ml IL-23 (high), or 5 µM R848. Recombinant human IL-17A (rhIL-17A) was used as positive control. Panels (B,C) display representative experiments out of two independent ones with similar results.

    Journal: Frontiers in Immunology

    Article Title: A Reappraisal on the Potential Ability of Human Neutrophils to Express and Produce IL-17 Family Members In Vitro: Failure to Reproducibly Detect It

    doi: 10.3389/fimmu.2018.00795

    Figure Lengend Snippet: Staining human neutrophils by anti-IL-17A (AF-317-NA) polyclonal antibodies (Abs). (A) Immunofluorescence (top panels) and immunohistochemistry (lower panels) stainings of two FFPE cases of human pustular psoriasis using anti-IL-17A (AF-317-NA) and anti-CD66b Abs (as labeled). Top panels show DAPI, FITC channel, and merge to recognize neutrophil shape; lower panels show different magnification of IHC and double IHC to characterize IL-17A + cells with the neutrophil marker CD66b. (B) Cytospins of neutrophils, either untreated (top panels) or treated with 5 µM R848 (bottom panels) for 3 h, were stained with anti-IL-17A (AF-317-NA, left panels) and anti-CXCL8 (right panels) Abs. Original magnification 200× [first row in (A) and left image in third row, scale bar 100 µm] and 400× [second row in (A) , center/right images in third row in (A) , as well as in (B) , scale bar 50 µm]. Images of the second row in (A) represent magnifications of images in first row. (C) AF-317-NA immunoblot of lysates from neutrophils either freshly isolated (T 0 , from two donors) or incubated for 3 h with or without 2 µg/ml IL-6 plus 0.2 µg/ml IL-23 (low), 20 µg/ml IL-6 plus 2 µg/ml IL-23 (high), or 5 µM R848. Recombinant human IL-17A (rhIL-17A) was used as positive control. Panels (B,C) display representative experiments out of two independent ones with similar results.

    Article Snippet: Cytokine Production Cytokine concentrations in cell-free supernatants and cell lysates were measured by commercial enzyme-linked immunosorbent (ELISA) kits, specific for: IL-17A (DY317 from R & D systems and 88-7176 from eBioscience), IL-17A/F (88-7117, eBioscience), IL-17B [ABKA2223 from Abnova (Taipei, Taiwan) and ab171344 from Abcam (Cambridge, United Kingdom)], IL-17F (887478, eBioscience), and CXCL8 (Mabtech, Nacka Strand, Sweden).

    Techniques: Staining, Immunofluorescence, Immunohistochemistry, Formalin-fixed Paraffin-Embedded, Labeling, Marker, Isolation, Incubation, Recombinant, Positive Control

    IL-17A, IL-17F, IL-17RA, IL-17RC, CXCL8, TNFα, and SOCS3 mRNA expression, as well as IL-17R surface expression, in neutrophils from patients with psoriasis. (A) Neutrophils isolated from healthy donors (HDs) ( n = 3) or psoriatic patients ( n = 3) were cultured for 20 h with 100 U/ml IFNγ plus 100 ng/ml LPS, 5 µM R848, or 500 ng/ml IL-17A to evaluate IL-17A, IL-17F, IL-17RA, IL-17RC, CXCL8, TNFα, and SOCS3 mRNA expression by reverse transcription quantitative real-time PCR. Gene expression data are depicted as mean normalized expression (MNE) units after GAPDH mRNA normalization. (B) Surface IL-17RA and IL-17RC expression evaluated by flow cytometry in human neutrophils from HDs or psoriatic patients. Values represent the mean ± SEM ( n = 3). For the data of panels (A,B) no significant differences between HDs or psoriatic patients were observed by two-way ANOVA followed by Bonferroni’s post-test.

    Journal: Frontiers in Immunology

    Article Title: A Reappraisal on the Potential Ability of Human Neutrophils to Express and Produce IL-17 Family Members In Vitro: Failure to Reproducibly Detect It

    doi: 10.3389/fimmu.2018.00795

    Figure Lengend Snippet: IL-17A, IL-17F, IL-17RA, IL-17RC, CXCL8, TNFα, and SOCS3 mRNA expression, as well as IL-17R surface expression, in neutrophils from patients with psoriasis. (A) Neutrophils isolated from healthy donors (HDs) ( n = 3) or psoriatic patients ( n = 3) were cultured for 20 h with 100 U/ml IFNγ plus 100 ng/ml LPS, 5 µM R848, or 500 ng/ml IL-17A to evaluate IL-17A, IL-17F, IL-17RA, IL-17RC, CXCL8, TNFα, and SOCS3 mRNA expression by reverse transcription quantitative real-time PCR. Gene expression data are depicted as mean normalized expression (MNE) units after GAPDH mRNA normalization. (B) Surface IL-17RA and IL-17RC expression evaluated by flow cytometry in human neutrophils from HDs or psoriatic patients. Values represent the mean ± SEM ( n = 3). For the data of panels (A,B) no significant differences between HDs or psoriatic patients were observed by two-way ANOVA followed by Bonferroni’s post-test.

    Article Snippet: Cytokine Production Cytokine concentrations in cell-free supernatants and cell lysates were measured by commercial enzyme-linked immunosorbent (ELISA) kits, specific for: IL-17A (DY317 from R & D systems and 88-7176 from eBioscience), IL-17A/F (88-7117, eBioscience), IL-17B [ABKA2223 from Abnova (Taipei, Taiwan) and ab171344 from Abcam (Cambridge, United Kingdom)], IL-17F (887478, eBioscience), and CXCL8 (Mabtech, Nacka Strand, Sweden).

    Techniques: Expressing, Isolation, Cell Culture, Real-time Polymerase Chain Reaction, Flow Cytometry, Cytometry

    Lack of IL-17A and IL-17F production by human neutrophils activated by IL-6 plus IL-23 in combination with inactivated Aspergillus fumigatus hyphae or conidia. Neutrophils (5 × 10 6 /ml) were incubated with or without 20 µg/ml IL-6 plus 2 µg/ml IL-23 and then cultured for three more hours in the presence or not of inactivated A. fumigatus conidia and hyphae (used at 1:5 and 1:1, respectively). After incubation, IL-17A (A) and CXCL8 (B) levels were determined in cell-free supernatants and in corresponding cell pellets by specific ELISA. Values are depicted as the mean ± SD or as not detected (nd) when values were under the detection limit ( n = 3). Asterisks stand for significant differences as compared to untreated cells: * P

    Journal: Frontiers in Immunology

    Article Title: A Reappraisal on the Potential Ability of Human Neutrophils to Express and Produce IL-17 Family Members In Vitro: Failure to Reproducibly Detect It

    doi: 10.3389/fimmu.2018.00795

    Figure Lengend Snippet: Lack of IL-17A and IL-17F production by human neutrophils activated by IL-6 plus IL-23 in combination with inactivated Aspergillus fumigatus hyphae or conidia. Neutrophils (5 × 10 6 /ml) were incubated with or without 20 µg/ml IL-6 plus 2 µg/ml IL-23 and then cultured for three more hours in the presence or not of inactivated A. fumigatus conidia and hyphae (used at 1:5 and 1:1, respectively). After incubation, IL-17A (A) and CXCL8 (B) levels were determined in cell-free supernatants and in corresponding cell pellets by specific ELISA. Values are depicted as the mean ± SD or as not detected (nd) when values were under the detection limit ( n = 3). Asterisks stand for significant differences as compared to untreated cells: * P

    Article Snippet: Cytokine Production Cytokine concentrations in cell-free supernatants and cell lysates were measured by commercial enzyme-linked immunosorbent (ELISA) kits, specific for: IL-17A (DY317 from R & D systems and 88-7176 from eBioscience), IL-17A/F (88-7117, eBioscience), IL-17B [ABKA2223 from Abnova (Taipei, Taiwan) and ab171344 from Abcam (Cambridge, United Kingdom)], IL-17F (887478, eBioscience), and CXCL8 (Mabtech, Nacka Strand, Sweden).

    Techniques: Incubation, Cell Culture, Enzyme-linked Immunosorbent Assay

    IL-17A, IL-17F, CXCL8, and IL-1ra mRNA expression levels in human neutrophils activated by a variety of stimuli. Human neutrophils were cultured at 5 × 10 6 /ml for up to 20 h with (A) 100 U/ml IFNγ and/or 100 ng/ml LPS; (B) 1,000 U/ml IFNα and/or 5 µM R848; (C) 10 ng/ml GM-CSF or 100 nM fMLF; (D) 1,000 U/ml G-CSF or 5 ng/ml TNFα. IL-17A, IL-17F, CXCL8, and IL-1ra mRNA expression was evaluated by reverse transcription quantitative real-time PCR (RT-qPCR) and data depicted as mean normalized expression (MNE) units after GAPDH mRNA normalization. The experiments depicted in each panels (A–D) are representative of at least three ones with similar results. Error bars stand for SEs calculated from triplicate qPCR reactions.

    Journal: Frontiers in Immunology

    Article Title: A Reappraisal on the Potential Ability of Human Neutrophils to Express and Produce IL-17 Family Members In Vitro: Failure to Reproducibly Detect It

    doi: 10.3389/fimmu.2018.00795

    Figure Lengend Snippet: IL-17A, IL-17F, CXCL8, and IL-1ra mRNA expression levels in human neutrophils activated by a variety of stimuli. Human neutrophils were cultured at 5 × 10 6 /ml for up to 20 h with (A) 100 U/ml IFNγ and/or 100 ng/ml LPS; (B) 1,000 U/ml IFNα and/or 5 µM R848; (C) 10 ng/ml GM-CSF or 100 nM fMLF; (D) 1,000 U/ml G-CSF or 5 ng/ml TNFα. IL-17A, IL-17F, CXCL8, and IL-1ra mRNA expression was evaluated by reverse transcription quantitative real-time PCR (RT-qPCR) and data depicted as mean normalized expression (MNE) units after GAPDH mRNA normalization. The experiments depicted in each panels (A–D) are representative of at least three ones with similar results. Error bars stand for SEs calculated from triplicate qPCR reactions.

    Article Snippet: Cytokine Production Cytokine concentrations in cell-free supernatants and cell lysates were measured by commercial enzyme-linked immunosorbent (ELISA) kits, specific for: IL-17A (DY317 from R & D systems and 88-7176 from eBioscience), IL-17A/F (88-7117, eBioscience), IL-17B [ABKA2223 from Abnova (Taipei, Taiwan) and ab171344 from Abcam (Cambridge, United Kingdom)], IL-17F (887478, eBioscience), and CXCL8 (Mabtech, Nacka Strand, Sweden).

    Techniques: Expressing, Cell Culture, Real-time Polymerase Chain Reaction, Quantitative RT-PCR

    Secretion of inflammation factors by dermal- and adipose-endothelial cells. Secretion of CXCL8, IL-6 and CCL2 after a 24 h exposure to 0, 40 or 100 µg/mL BWE. Basal amounts of protein in culture medium containing 100 µg/mL BWE without cells: CXCL8:

    Journal: International Journal of Molecular Sciences

    Article Title: Burn Eschar Stimulates Fibroblast and Adipose Mesenchymal Stromal Cell Proliferation and Migration but Inhibits Endothelial Cell Sprouting

    doi: 10.3390/ijms18081790

    Figure Lengend Snippet: Secretion of inflammation factors by dermal- and adipose-endothelial cells. Secretion of CXCL8, IL-6 and CCL2 after a 24 h exposure to 0, 40 or 100 µg/mL BWE. Basal amounts of protein in culture medium containing 100 µg/mL BWE without cells: CXCL8:

    Article Snippet: IL-6 and CCL2 (both R & D Systems, Abingdon, UK) and CXCL8 (Sanquin, Amsterdam, The Netherlands).

    Techniques:

    Alleviation of resistance via blockade of IL-8 signaling. Cytotoxicity of PC9 and HCC827 cells mediated by NK cells or TRAIL, as indicated. Tumor cells were left untreated, treated with erlotinib for 3 days, or treated with erlotinib for 3 days followed by exposure to anti-IL-8 neutralizing antibody before the assay

    Journal: Cell Death & Disease

    Article Title: Short-term EGFR blockade enhances immune-mediated cytotoxicity of EGFR mutant lung cancer cells: rationale for combination therapies

    doi: 10.1038/cddis.2016.297

    Figure Lengend Snippet: Alleviation of resistance via blockade of IL-8 signaling. Cytotoxicity of PC9 and HCC827 cells mediated by NK cells or TRAIL, as indicated. Tumor cells were left untreated, treated with erlotinib for 3 days, or treated with erlotinib for 3 days followed by exposure to anti-IL-8 neutralizing antibody before the assay

    Article Snippet: A third group consisted of tumor cells treated with erlotinib for 72 h followed by treatment with a commercially available neutralizing anti-IL-8 antibody (MAB208, R & D Systems, Minneapolis, MN, USA; 10 μ g/ml) for 96 h. Cells were labeled and used as targets with NK effector cells or recombinant TRAIL.

    Techniques:

    Human melanoma tumors express ligands for CXCR2, but tumor-infiltrating T cells lack CXCR2 expression. (A) Paraffin-embedded, melanoma lymph node metastases were analyzed by immunohistochemical staining for chemokine CXCL8 and CXCL1. Representative staining is shown at 40X magnification. (B) CXCR2 expression on PBMCs, as determined by flow cytometry. PBMCs from five healthy donors were stained with anti-CD3 and anti-CXCR2. (C) FACS analysis of CXCR2 expression on tumor-infiltrating lymphocytes (TILs). TILs isolated from four melanoma patients were stained with anti-CD4, CD8 and CXCR2. Representative results of all stained samples were shown.

    Journal: Clinical cancer research : an official journal of the American Association for Cancer Research

    Article Title: Transduction of Tumor-Specific T Cells with CXCR2 Chemokine Receptor Improves Migration to Tumor and Antitumor Immune Responses

    doi: 10.1158/1078-0432.CCR-10-0712

    Figure Lengend Snippet: Human melanoma tumors express ligands for CXCR2, but tumor-infiltrating T cells lack CXCR2 expression. (A) Paraffin-embedded, melanoma lymph node metastases were analyzed by immunohistochemical staining for chemokine CXCL8 and CXCL1. Representative staining is shown at 40X magnification. (B) CXCR2 expression on PBMCs, as determined by flow cytometry. PBMCs from five healthy donors were stained with anti-CD3 and anti-CXCR2. (C) FACS analysis of CXCR2 expression on tumor-infiltrating lymphocytes (TILs). TILs isolated from four melanoma patients were stained with anti-CD4, CD8 and CXCR2. Representative results of all stained samples were shown.

    Article Snippet: Immunohistochemistry was performed on paraffin-embedded melanoma tumor tissue sections. mAbs against human CXCL1 (Proteintech) and CXCL8 (Santa Cruz biotechnology) were used.

    Techniques: Expressing, Immunohistochemistry, Staining, Flow Cytometry, Cytometry, FACS, Isolation

    Serial changes in plasma cytokine/chemokine concentrations during the course of hospitalization. There was sustained elevation of the proinflammatory cytokines (IL-6, CXCL8/IL-8, CCL2/MCP-1, sTNFR-1) in severe pH1N1 pneumonia; the adaptive-immunity related cytokines (CXCL10/IP-10, CXCL9/MIG, IL-17A) were markedly suppressed compared with seasonal influenza. All patients with pH1N1 influenza (severe pneumonia, n = 34; milder illness, n = 29) received antiviral treatment soon after hospitalization/recruitment; none had received high-dose corticosteroids or other immunosuppressants for ‘viral pneumonitis’ or ‘ARDS’ [8] . Among seasonal influenza patients (most had complicated illnesses, see Table 1 footnotes), 30(57%) received antiviral treatment. Median concentrations at each time point are shown for each group; the interquartile ranges (presented in Table 1 ) are omitted here for clarity. Fewer mild pH1N1 and untreated seasonal influenza patients remained hospitalized at day 6–7 for study (Day 1, n = 116; Day 3–4, n = 62; Day 6–7, n = 30).

    Journal: PLoS ONE

    Article Title: Cytokine Response Patterns in Severe Pandemic 2009 H1N1 and Seasonal Influenza among Hospitalized Adults

    doi: 10.1371/journal.pone.0026050

    Figure Lengend Snippet: Serial changes in plasma cytokine/chemokine concentrations during the course of hospitalization. There was sustained elevation of the proinflammatory cytokines (IL-6, CXCL8/IL-8, CCL2/MCP-1, sTNFR-1) in severe pH1N1 pneumonia; the adaptive-immunity related cytokines (CXCL10/IP-10, CXCL9/MIG, IL-17A) were markedly suppressed compared with seasonal influenza. All patients with pH1N1 influenza (severe pneumonia, n = 34; milder illness, n = 29) received antiviral treatment soon after hospitalization/recruitment; none had received high-dose corticosteroids or other immunosuppressants for ‘viral pneumonitis’ or ‘ARDS’ [8] . Among seasonal influenza patients (most had complicated illnesses, see Table 1 footnotes), 30(57%) received antiviral treatment. Median concentrations at each time point are shown for each group; the interquartile ranges (presented in Table 1 ) are omitted here for clarity. Fewer mild pH1N1 and untreated seasonal influenza patients remained hospitalized at day 6–7 for study (Day 1, n = 116; Day 3–4, n = 62; Day 6–7, n = 30).

    Article Snippet: Plasma concentrations of 10 cytokines/chemokines including interleukin(IL)-1β, IL-6, IL-10, IL-12p70, tumour necrosis factor(TNF)-α, CXCL8/IL-8, CXCL9/MIG (monokine induced by interferon-γ), CXCL10/IP-10 (interferon gamma-induced protein-10), CCL2/MCP-1 (monocyte chemoattractant protein-1), and CCL5/RANTES, were assayed using cytometric bead array (CBA) reagents (BD Pharmingen, San Diego,CA) with four-color FACSCalibur flow-cytometer (BD Biosciences Corp, San Jose,CA) as previously described , .

    Techniques:

    CXCL8 expression via NF-κB activation in SL-treated THP-1 cells. THP-1 cells were pretreated with α2,3- or α2,6-SL (1 or 10 μM) for 3 h and then treated with LPS (0.1 μg/ml) or PAK (MOI of 50). (A) CXCL8 mRNA levels were analyzed by RT-PCR. (B and E) CXCL8 secretion was analyzed by ELISA. (C) IκBα degradation was analyzed by Western blotting. (D) THP-1 cells were pretreated with the NF-κB inhibitor BAY11-7082 (10 μM) and then treated with α2,3- or α2,6-SL (1 or 10 μM) and PAK (MOI of 50) as previously described. CXCL8 expression was analyzed by RT-PCR or ELISA. Statistical significance was determined by ANOVA (Tukey’s test). ANOVA test results are shown as letters above columns. Means not sharing the same letter are significantly different. GAPDH, glyceraldehyde 3-phosphate dehydrogenase.

    Journal: Infection and Immunity

    Article Title: Bacterial Clearance Is Enhanced by α2,3- and α2,6-Sialyllactose via Receptor-Mediated Endocytosis and Phagocytosis

    doi: 10.1128/IAI.00694-18

    Figure Lengend Snippet: CXCL8 expression via NF-κB activation in SL-treated THP-1 cells. THP-1 cells were pretreated with α2,3- or α2,6-SL (1 or 10 μM) for 3 h and then treated with LPS (0.1 μg/ml) or PAK (MOI of 50). (A) CXCL8 mRNA levels were analyzed by RT-PCR. (B and E) CXCL8 secretion was analyzed by ELISA. (C) IκBα degradation was analyzed by Western blotting. (D) THP-1 cells were pretreated with the NF-κB inhibitor BAY11-7082 (10 μM) and then treated with α2,3- or α2,6-SL (1 or 10 μM) and PAK (MOI of 50) as previously described. CXCL8 expression was analyzed by RT-PCR or ELISA. Statistical significance was determined by ANOVA (Tukey’s test). ANOVA test results are shown as letters above columns. Means not sharing the same letter are significantly different. GAPDH, glyceraldehyde 3-phosphate dehydrogenase.

    Article Snippet: Ninety-six-well microtiter plates were coated with anti-CXCL8 antibodies (dilution, 1:250; BD Biosciences) at 4°C overnight and then blocked with 2% BSA for 1 h at room temperature, followed by the addition of samples.

    Techniques: Expressing, Activation Assay, Reverse Transcription Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Western Blot

    Clathrin-TRIF-dependent NF-κB activation in SL-treated THP-1 cells. (A) Schematic of the experimental design. (B) THP-1 cells were transfected with clathrin siRNA, pretreated with α2,3- or α2,6-SL (10 μM) for 3 h, and then treated with PAK (MOI of 50). IκBα degradation was analyzed by Western blotting, and CXCL8 secretion was analyzed by ELISA. Statistical significance was determined by Student's t test. P values were

    Journal: Infection and Immunity

    Article Title: Bacterial Clearance Is Enhanced by α2,3- and α2,6-Sialyllactose via Receptor-Mediated Endocytosis and Phagocytosis

    doi: 10.1128/IAI.00694-18

    Figure Lengend Snippet: Clathrin-TRIF-dependent NF-κB activation in SL-treated THP-1 cells. (A) Schematic of the experimental design. (B) THP-1 cells were transfected with clathrin siRNA, pretreated with α2,3- or α2,6-SL (10 μM) for 3 h, and then treated with PAK (MOI of 50). IκBα degradation was analyzed by Western blotting, and CXCL8 secretion was analyzed by ELISA. Statistical significance was determined by Student's t test. P values were

    Article Snippet: Ninety-six-well microtiter plates were coated with anti-CXCL8 antibodies (dilution, 1:250; BD Biosciences) at 4°C overnight and then blocked with 2% BSA for 1 h at room temperature, followed by the addition of samples.

    Techniques: Activation Assay, Transfection, Western Blot, Enzyme-linked Immunosorbent Assay

    Effect of nitric oxide addition on binding and dissociation of the LDV-FITC probe on U937 cells, treated with different Gα i -coupled receptor ligands . LDV-FITC probe binding and dissociation on U937 cells stably transfected with different GPCRs plotted as mean channel fluorescence (MCF) versus time. A, The experiment involved sequential addition of fluorescent LDV-FITC probe (4 nM, below saturation, added 2 min prior to addition of Gαi-coupled receptor ligand, fMLFF, 100 nM), and different concentrations of DEA-NONOate (nitric oxide donor). Dashed line indicates the non-specific binding of the LDV-FITC probe determined using an excess of unlabelled LDV competitor (as shown in Fig. 2D,E). B, The experiment involved sequential addition of fluorescent LDV-FITC probe (4 nM), CXCL12/SDF-1 (12 nM), and DEA-NONOate (250 μM, nitric oxide donor) or vehicle (control). Rapid and reversible binding of the probe reflects the VLA-4 affinity change [14]. C, The experiment involved sequential addition of the fluorescent LDV-FITC probe (4 nM), CXCL8/IL-8 (20 nM), and DEA-NONOate (250 μM, nitric oxide donor) or vehicle (control). D, The experiment involved sequential addition of the DEA-NONOate (250 μM, nitric oxide donor) or vehicle (control) at the 0 time point, and the fluorescent LDV-FITC probe (4 nM), CXCL12/SDF-1 (12 nM). Excess unlabelled competitor LDV (1 μM) is added at the end of the experiment to determine the non-specific binding of the probe (panels D, and E). E, The experiment involved sequential addition of DEA-NONOate (250 μM, nitric oxide donor) or vehicle (control) at the 0 time point, and the fluorescent LDV-FITC probe (4 nM), CXCL8/IL-8 (20 nM) (arrows). According to the unpaired t test, the means are significantly different (p

    Journal: BMC Immunology

    Article Title: Nitric oxide/cGMP pathway signaling actively down-regulates ?4?1-integrin affinity: an unexpected mechanism for inducing cell de-adhesion

    doi: 10.1186/1471-2172-12-28

    Figure Lengend Snippet: Effect of nitric oxide addition on binding and dissociation of the LDV-FITC probe on U937 cells, treated with different Gα i -coupled receptor ligands . LDV-FITC probe binding and dissociation on U937 cells stably transfected with different GPCRs plotted as mean channel fluorescence (MCF) versus time. A, The experiment involved sequential addition of fluorescent LDV-FITC probe (4 nM, below saturation, added 2 min prior to addition of Gαi-coupled receptor ligand, fMLFF, 100 nM), and different concentrations of DEA-NONOate (nitric oxide donor). Dashed line indicates the non-specific binding of the LDV-FITC probe determined using an excess of unlabelled LDV competitor (as shown in Fig. 2D,E). B, The experiment involved sequential addition of fluorescent LDV-FITC probe (4 nM), CXCL12/SDF-1 (12 nM), and DEA-NONOate (250 μM, nitric oxide donor) or vehicle (control). Rapid and reversible binding of the probe reflects the VLA-4 affinity change [14]. C, The experiment involved sequential addition of the fluorescent LDV-FITC probe (4 nM), CXCL8/IL-8 (20 nM), and DEA-NONOate (250 μM, nitric oxide donor) or vehicle (control). D, The experiment involved sequential addition of the DEA-NONOate (250 μM, nitric oxide donor) or vehicle (control) at the 0 time point, and the fluorescent LDV-FITC probe (4 nM), CXCL12/SDF-1 (12 nM). Excess unlabelled competitor LDV (1 μM) is added at the end of the experiment to determine the non-specific binding of the probe (panels D, and E). E, The experiment involved sequential addition of DEA-NONOate (250 μM, nitric oxide donor) or vehicle (control) at the 0 time point, and the fluorescent LDV-FITC probe (4 nM), CXCL8/IL-8 (20 nM) (arrows). According to the unpaired t test, the means are significantly different (p

    Article Snippet: Human recombinant CXCL12/SDF-1α, and recombinant human CXCL8/IL-8 were from R & D Systems.

    Techniques: Binding Assay, Stable Transfection, Transfection, Fluorescence

    Effect of guanylyl cyclase activator on binding and dissociation of the LDV-FITC probe on U937 cells, treated with different Gα i -coupled receptor ligands . LDV-FITC probe binding and dissociation on U937 cells stably transfected with different GPCRs plotted as mean channel fluorescence (MCF) versus time. A, The experiment involved sequential addition of the fluorescent LDV-FITC probe (4 nM, below saturation, added 2 min prior to addition of the Gαi-coupled receptor ligand, fMLFF, 100 nM), and different concentrations of BAY 41-2272 (guanylyl cyclase activator). B, The experiment involved sequential addition of the BAY 41-2272 (100 μM, guanylyl cyclase activator) or vehicle (control) at the 0 time point, and the fluorescent LDV-FITC probe (4 nM), and CXCL12/SDF-1 (12 nM). C, The experiment involved sequential addition of BAY 41-2272 (100 μM, guanylyl cyclase activator) or vehicle (control) at the 0 time point, and the fluorescent LDV-FITC probe (4 nM), and CXCL8/IL-8 (20 nM). The means are significantly different (p

    Journal: BMC Immunology

    Article Title: Nitric oxide/cGMP pathway signaling actively down-regulates ?4?1-integrin affinity: an unexpected mechanism for inducing cell de-adhesion

    doi: 10.1186/1471-2172-12-28

    Figure Lengend Snippet: Effect of guanylyl cyclase activator on binding and dissociation of the LDV-FITC probe on U937 cells, treated with different Gα i -coupled receptor ligands . LDV-FITC probe binding and dissociation on U937 cells stably transfected with different GPCRs plotted as mean channel fluorescence (MCF) versus time. A, The experiment involved sequential addition of the fluorescent LDV-FITC probe (4 nM, below saturation, added 2 min prior to addition of the Gαi-coupled receptor ligand, fMLFF, 100 nM), and different concentrations of BAY 41-2272 (guanylyl cyclase activator). B, The experiment involved sequential addition of the BAY 41-2272 (100 μM, guanylyl cyclase activator) or vehicle (control) at the 0 time point, and the fluorescent LDV-FITC probe (4 nM), and CXCL12/SDF-1 (12 nM). C, The experiment involved sequential addition of BAY 41-2272 (100 μM, guanylyl cyclase activator) or vehicle (control) at the 0 time point, and the fluorescent LDV-FITC probe (4 nM), and CXCL8/IL-8 (20 nM). The means are significantly different (p

    Article Snippet: Human recombinant CXCL12/SDF-1α, and recombinant human CXCL8/IL-8 were from R & D Systems.

    Techniques: Binding Assay, Stable Transfection, Transfection, Fluorescence