Journal: PLoS Pathogens

Article Title: Plasminogen Controls Inflammation and Pathogenesis of Influenza Virus Infections via Fibrinolysis

doi: 10.1371/journal.ppat.1003229

Figure Lengend Snippet: (A) Histopathological analysis of lungs from infected WT and PLG-KO mice inoculated with A/PR/8/34 virus (day 3 post-infection) or A/Netherlands/602/09 virus (day 5 post-infection). Thin sections of lungs obtained from infected and uninfected WT and PLG-KO mice (as indicated) were stained with hematoxilin end eosin (HE) to evaluate histopathological changes. Note the marked infiltration of inflammatory cells in the lungs of infected WT mice, which was largely absent in the lungs of PLG-KO mice. The results shown are representative for two-three mice for both groups. Immunohistochemistry (IHC) using a monoclonal antibody for the influenza A virus nucleoprotein was used to detect virus-infected cells. Cells positive for the presence of viral antigen stained red. (B) Cytokine levels in BAL were assessed by ELISA on the indicated days post inoculation of WT (black bars) and PLG-KO mice (white bars) with IAV A/PR/8/34 or A/Netherlands/602/09. Data represent mean ± s.e.m. of 3–6 mice per group.

Article Snippet: Viruses, cells, and reagents used, were: IAV A/Netherlands/602/09 , A/chicken/Ivory-Coast/1787/2006 , A/PR/8/34 (American Type Culture Collection, ATCC), A549 cells (ATCC), Madin-Darby Canine Kidney cells (MDCK, ATCC), trypsin (Becton Dickinson), plasminogen and 6-AHA (Sigma), Ancrod (NIBSC), 23-Plex Mouse Cytokine Assay (Bio-Rad), ELISA kits for mouse -IL-6, -KC, -–RANTES, -IFN-α -IFN-γ(R&D Systems), -plasminogen (Mybiosource), -active plasmin (Kordia), -D-dimer, -fibrinogen and -FDP (Genway), antibodies anti-HA (Santa Cruz), anti-tubulin (Sigma), anti-NP (ATCC), anti-fibrinogen (Genway).

Techniques: Infection, Virus, Staining, Immunohistochemistry, Enzyme-linked Immunosorbent Assay

Journal: PLoS Pathogens

Article Title: Plasminogen Controls Inflammation and Pathogenesis of Influenza Virus Infections via Fibrinolysis

doi: 10.1371/journal.ppat.1003229

Figure Lengend Snippet: (A) Levels of Plasminogen, Active Plasmin, FDP, D-dimer and Fibrinogen, were determined by ELISA in the BAL of A/PR/8/34 infected or uninfected (−) C57BL/6 mice after the indicated days post-inoculation. Markers were also evaluated in the BAL of WT or PLG-KO mice infected with A/Netherlands/602/09 virus. Data represent mean ± s.e.m of n = 3–6 mice per group. (B) Western blot analysis for the detection of fibrinogen and FDP in the lungs of IAV-infected mice on the indicated days post inoculation (representative of n = 3). kDa: apparent molecular weight. n = mice per group. (C) Presence of fibrinogen was assessed in the blood of mice treated or not with Ancrod by ELISA (left panel) or Western blot analysis (right panel). The results represent the mean values ± s.e.m from 3 individual animals per group for the ELISA. The western blot analysis is representative for results of 3 mice per group.

Article Snippet: Viruses, cells, and reagents used, were: IAV A/Netherlands/602/09 , A/chicken/Ivory-Coast/1787/2006 , A/PR/8/34 (American Type Culture Collection, ATCC), A549 cells (ATCC), Madin-Darby Canine Kidney cells (MDCK, ATCC), trypsin (Becton Dickinson), plasminogen and 6-AHA (Sigma), Ancrod (NIBSC), 23-Plex Mouse Cytokine Assay (Bio-Rad), ELISA kits for mouse -IL-6, -KC, -–RANTES, -IFN-α -IFN-γ(R&D Systems), -plasminogen (Mybiosource), -active plasmin (Kordia), -D-dimer, -fibrinogen and -FDP (Genway), antibodies anti-HA (Santa Cruz), anti-tubulin (Sigma), anti-NP (ATCC), anti-fibrinogen (Genway).

Techniques: Enzyme-linked Immunosorbent Assay, Infection, Virus, Western Blot, Molecular Weight

Journal: PLoS Pathogens

Article Title: Plasminogen Controls Inflammation and Pathogenesis of Influenza Virus Infections via Fibrinolysis

doi: 10.1371/journal.ppat.1003229

Figure Lengend Snippet: (A) Survival and weight loss of mice treated with Ancrod (open symbols, n = 11) or not (closed symbols, n = 11) after infection with IAV A/PR/8/34 (squares) or uninfected mice (diamonds, n = 5). Weight loss data represent weight average ± s.e.m of the above indicated number of mice. (B) Cytokines levels in the BAL were measured by ELISA after A/PR/8/34 infection of WT and PLG-KO (KO) mice treated with Ancrod (white bars) or not (black bars). Data represent mean ± s.e.m. of n = 4 mice per group. (C) Survival rate (left panels) and weight loss (right panels) of WT (squares) and PLG-KO (triangles) mice treated with Ancrod (open symbols) or not (closed symbols) after intranasal inoculation with IAV A/PR/8/34 (n = 8–10 mice per group). Weight loss data represent weight average ± s.e.m of the above indicated number of mice.

Article Snippet: Viruses, cells, and reagents used, were: IAV A/Netherlands/602/09 , A/chicken/Ivory-Coast/1787/2006 , A/PR/8/34 (American Type Culture Collection, ATCC), A549 cells (ATCC), Madin-Darby Canine Kidney cells (MDCK, ATCC), trypsin (Becton Dickinson), plasminogen and 6-AHA (Sigma), Ancrod (NIBSC), 23-Plex Mouse Cytokine Assay (Bio-Rad), ELISA kits for mouse -IL-6, -KC, -–RANTES, -IFN-α -IFN-γ(R&D Systems), -plasminogen (Mybiosource), -active plasmin (Kordia), -D-dimer, -fibrinogen and -FDP (Genway), antibodies anti-HA (Santa Cruz), anti-tubulin (Sigma), anti-NP (ATCC), anti-fibrinogen (Genway).

Techniques: Infection, Enzyme-linked Immunosorbent Assay

Journal: bioRxiv

Article Title: Antimitotic chemotherapy promotes tumor NF-kB secretory phenotype and immunosuppressive CXCR2+ neutrophils chemotaxis in triple-negative breast cancers

doi: 10.1101/2024.12.21.629876

Figure Lengend Snippet: A: PCA analysis of 4T1-Luc2 CRISPR CTL and CRISPR STING treated or not by paclitaxel and analyzed by 3’Seq RNA Profiling. B, C: Volcano plots representing differentially up- and downregulated expressed under paclitaxel treatment in 4T1-Luc2 CRISPR CTL and CRISPR STING , respectively. D, E: Signaling pathway signatures up- or downregulated by paclitaxel in 4T1-Luc2 CRISPR CTL and CRISPR STING , respectively. F, G: Concentration in CCL5 and CXCL1 by ELISA in the culture medium of 4T1-Luc2 CRISPR CTL and CRISPR STING treated or not by paclitaxel. H: Concentration in CXCL1 by ELISA spot in the culture medium of paclitaxel-treated 4T1-Luc2 CRISPR CTL and CRISPR STING under treatment with AS602868 or not.

Article Snippet: For ELISA assays, levels of different chemokines in cell supernatants were determined according to the manufacturer’s protocols using following kits: CXCL1 (Mouse CXCL1 Sandwich ELISA Kit, Proteintech) and CCL5 (Mouse CCL5 Sandwich ELISA Kit, Proteintech).

Techniques: CRISPR, Concentration Assay, Enzyme-linked Immunosorbent Assay

Journal: bioRxiv

Article Title: Antimitotic chemotherapy promotes tumor NF-kB secretory phenotype and immunosuppressive CXCR2+ neutrophils chemotaxis in triple-negative breast cancers

doi: 10.1101/2024.12.21.629876

Figure Lengend Snippet: A: Impact of STING on paclitaxel-induced transcriptional expression (3’SRP) of Cxcl1, Cxcl5, Ccl5 and Ccl20 comparing fold-change expression (paclitaxel versus untreated) in both CRISPR CTL and CRISPR STING . B: Transcriptional expression fold change of Cxcl1, Cxcl5, Ccl5 and Ccl20 in CRISPR CTL and CRISPR STING under paclitaxel or not. C : Expression of CXCL1 and CCL5 by human TNBC cells using scRNAseq analysis from Broad Institute ( https://singlecell.broadinstitute.org/single_cell/study/SCP1106/stromal-cell-diversity-associated-with-immune-evasion-in-human-triple-negative-breast-cancer ).

Article Snippet: For ELISA assays, levels of different chemokines in cell supernatants were determined according to the manufacturer’s protocols using following kits: CXCL1 (Mouse CXCL1 Sandwich ELISA Kit, Proteintech) and CCL5 (Mouse CCL5 Sandwich ELISA Kit, Proteintech).

Techniques: Expressing, CRISPR

Journal: Nature communications

Article Title: Asthma reduces glioma formation by T cell decorin-mediated inhibition of microglia.

doi: 10.1038/s41467-021-27455-6

Figure Lengend Snippet: Fig. 3 Asthma reduces T cell induction of Ccl5 in microglia. a Schematic representation of the mouse Nf1OPG neuron-immune-cancer cell axis in the setting of asthma. b No difference in optic nerve Mdk RNA expression was observed between OVA- (n = 3) or HDM- (n = 3) treated mice and controls (n = 6). Bar graphs represent the means ± SEM of independent biological samples. One-way ANOVA with Bonferroni post hoc correction. c Ccl4 levels are similarly induced with increasing midkine concentrations (0–100 ng/ml) in CD3+ T cells from OVA- and PBS-treated mice. Data are presented as the means ± SEM of n = 3 independent biological samples. One-way ANOVA with Bonferroni post hoc correction. Data are presented as the means ± SEM. d, e Similarly increased CD8+, relative to CD4+, T cell content was detected in the optic nerves of Nf1OPG mice treated with PBS (n = 8), OVA (n = 8) or HDM (n = 8). One-way ANOVA with Bonferroni post hoc correction. Activated T cell medium from (f) OVA- (n = 4) and g HDM- (n = 4) treated mice induced lower levels of microglial Ccl5 relative to PBS-treated Nf1OPG mice. Two-tailed Student’s t test. Similar reductions of Ccl5 expression were observed in CD4+ and CD8+ T cells from (h) OVA- (n = 3) and (i) HDM- (n = 3) treated mice relative to PBS-treated controls (n = 3). One-way ANOVA with Bonferroni post hoc correction. Data are presented as the means ± SEM. d Scale bars, 40 µm. From left to right in each panel: b ns; c ns; ns; ns; ns; e P < 0.0001, P = 0.0022, P = 0.0011; f P = 0.0008; g P = 0.0015; h P = 0.0186, P = 0.0054; i P = 0.0224, P = 0.0007.

Article Snippet: In total, 5 × 105 microglia were grown in T cell-conditioned media (TCM) for 48 h8,9, followed Ccl5 (R&D Systems, MMR00) and decorin (Abcam, ab155454) determinations by ELISA.

Techniques: RNA Expression, Two Tailed Test, Expressing

Journal: BMC cancer

Article Title: Exercise accelerates recruitment of CD8 + T cell to promotes anti-tumor immunity in lung cancer via epinephrine.

doi: 10.1186/s12885-024-12224-7

Figure Lengend Snippet: Fig. 3 Transcriptional targets were examined by RNA-seq in tumors after exercise. a The number of differentially expressed genes in the Ex and Ctrl mice were detected by RNA-seq. b Volcano plot of differentially expressed genes in Ex compared with Ctrl tumor tissues. Red dots represent upregu lated genes, and blue dots represent downregulated genes. c Heat map of the differentially expressed genes in Ex compared with Ctrl tumor tissues. d Pathway enrichment analysis of significantly upregulated or downregulated genes. e-j RT-qPCR validates the gene expression of Cxcl10, Cxcl12, Cxcl14, Ppbp, Pf4 and Ccl8 in the tumors each group (n = 3) after tumor inoculation. k-l The expression levels of Cxcl10 and Cxcl12 were detected in the tumors in each group by ELISA (n = 8). The results of e-l are presented as mean ± SEM. Statistical analysis was performed using two-tailed unpaired t tests. *p < 0.05, **p < 0.01, ***p < 0.001

Article Snippet: The concentrations of Ccl5, Cxcl10 and Cxcl12 were measured in Lewis lung cancercellsculture supernatants and in the serum of mice using Ccl5 ELISA kit (Cusabio, CSBE09256m), Cxcl10 ELISA kit (Cusabio, CSB-E08183m) and Cxcl12 (Cusabio, CSB-E04723m).

Techniques: RNA Sequencing, Quantitative RT-PCR, Gene Expression, Expressing, Enzyme-linked Immunosorbent Assay, Two Tailed Test

Journal: BMC cancer

Article Title: Exercise accelerates recruitment of CD8 + T cell to promotes anti-tumor immunity in lung cancer via epinephrine.

doi: 10.1186/s12885-024-12224-7

Figure Lengend Snippet: Fig. 4 Exercise promotes CD8+ T cell recruitment with the assistance of Ccl5 and Cxcl10. a-d Correlation between Ccl5, Cxcl9, Cxcl10, and Cxcl11 mRNA level and CD8+ T cell using QUANTISEQ. Data were obtained from TIMER 2.0 (http://timer.comp-genomics.org/). e-h RT-qPCR analyses of mRNA expres sion of chemokines in the tumors of Ctrl and Ex groups (n = 3). i-j ELISA analysis of serum levels of Cxcl10 and Ccl5 in Ctrl and Ex mice (n = 8). k-m RT-qPCR analyses of mRNA expression of cytokines and PD-L1 in the tumors of Ctrl and Ex mice (n = 3). n Western blot analysis the expression of PD-L1 in the tu mors of Ctrl and Ex mice (n = 3). Full-length blots/gels are presented in Fig. 4n of source data. o Automatic quantification of PD-L1 IHC staining in indicated mice tumors (n = 8). p IHC staining of PD-L1 on tumor sections from different groups (n = 8). q Western blot analysis the expression of P53 in the tumors of Ctrl and Ex mice (n = 3). Full-length blots/gels are presented in Fig. 4q of source data. The results of e-m and o are presented as mean ± SEM. Statistical analysis was performed using two-tailed unpaired t tests. *p < 0.05, **p < 0.01, ***p < 0.001

Article Snippet: The concentrations of Ccl5, Cxcl10 and Cxcl12 were measured in Lewis lung cancercellsculture supernatants and in the serum of mice using Ccl5 ELISA kit (Cusabio, CSBE09256m), Cxcl10 ELISA kit (Cusabio, CSB-E08183m) and Cxcl12 (Cusabio, CSB-E04723m).

Techniques: Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Expressing, Western Blot, Immunohistochemistry, Two Tailed Test

Journal: BMC cancer

Article Title: Exercise accelerates recruitment of CD8 + T cell to promotes anti-tumor immunity in lung cancer via epinephrine.

doi: 10.1186/s12885-024-12224-7

Figure Lengend Snippet: Fig. 5 Exercise inhibits tumor progression by upregulation of EPI levels. a Serum levels of EPI in Ctrl, Ex and Ex-post groups were detected by ELISA in tumor-bearing mice (n = 8). b-d EPI mice receiving daily injections of EPI (0.5 mg/kg i.p.) (n = 8) and TE mice with daily running for 14 days after tumor inoculation (n = 8) compared with TC mice (n = 8). b Representative images. c Tumor weights of indicated groups. d Tumor volumes of different groups. e IHC staining of CD8, CD4, CD3 and GZMB on tumor sections from indicated groups (n = 8). f Automatic quantification of CD8, CD4, CD3 and GZMB IHC staining in indicated mice tumors (n = 8). Data are presented as the mean ± SEM in each group. Statistical analysis of a was performed using two-tailed unpaired t tests. c, d and f were performed using one-way analysis of variance (ANOVA). *p < 0.05, **p < 0.01, ***p < 0.001

Article Snippet: The concentrations of Ccl5, Cxcl10 and Cxcl12 were measured in Lewis lung cancercellsculture supernatants and in the serum of mice using Ccl5 ELISA kit (Cusabio, CSBE09256m), Cxcl10 ELISA kit (Cusabio, CSB-E08183m) and Cxcl12 (Cusabio, CSB-E04723m).

Techniques: Enzyme-linked Immunosorbent Assay, Immunohistochemistry, Two Tailed Test