Journal: Cell communication and signaling : CCS

Article Title: Helicobacter pylori disrupts gastric mucosal homeostasis by stimulating macrophages to secrete CCL3.

doi: 10.1186/s12964-024-01627-5

Figure Lengend Snippet: Fig. 3 H. pylori infection promotes the expression of CCL3. A Analysis chemokines in the medium of THP-1 co-cultured with H. pylori. B Differential analysis of chemokines about the GSE27411 dataset. C, D The protein and RNA expression level of CCL3 in THP-1 co-cultured with H. pylori. E The secretion level of CCL3 in the medium of H. pylori -infected macrophages. F The protein expression level of CCL3 in H. pylori infected mice gastric tissue (PBS: n = 2; H. pylori: n = 3). G The RNA expression level of CCL3 in H. pylori infected mice gastric tissue (PBS: n = 3; H. pylori: n = 5). H IHC analysis CCL3 expression in mice (PBS: n = 3; H. pylori: n = 6). I IHC analysis CCL3 expression in human (each group: n = 4). J The expression level of CCL3 in mice serum (each group: n = 4). K The expression level of CCL3 in human serum (each group: n = 3). Abbreviations: IHC, immunohistochemical. The data are presented as the mean ± S.D. after triplicate. Two groups were compared by t-test, multiple groups were compared by one-way analysis of variance (ANOVA). Note: *P < 0.05, **P < 0.01, ***P < 0.001

Article Snippet: Human CCL3 ELISA kit was purchased from CUSABIO (CSB-E04662h, China).

Techniques: Infection, Expressing, Cell Culture, RNA Expression, Immunohistochemical staining

Journal: Cell communication and signaling : CCS

Article Title: Helicobacter pylori disrupts gastric mucosal homeostasis by stimulating macrophages to secrete CCL3.

doi: 10.1186/s12964-024-01627-5

Figure Lengend Snippet: Fig. 5 CCL3 neutralizing antibody and CCL3 receptor inhibitor improve mucosal damage and inflammatory response. A Effect of CCL3 neutralizing antibody (100 ng/ml) on TEER of MKN28 cells. B The protein expression level of the tight junction in GES-1 cells treated with CCL3 neutralizing antibody in conditioned media. C IF detected tight junction between MKN28 cells treated with CCL3 neutralizing antibody in conditioned media. DAPI was used for nuclear staining (blue), Occludin was used for tight junction staining (green), arrows showed the disruption of tight junctions between cells. D, E Apoptosis and proliferation of GES-1 treated with CCL3 neutralizing antibody in conditioned media (D: Annexin V- PI apoptosis flow cytometry; E: EdU). F Effect of CCL3 receptor inhibitor (Maraviroc, 100 nM) on TEER of MKN28 cells. G The protein expression level of the tight junction in GES-1 cells treated with CCL3 receptor inhibitor. H IF detected the tight junction between MKN28 cells treated with CCL3 receptor inhibitor. I, J Apoptosis and proliferation of GES-1 treated with CCL3 receptor inhibitor (I: Annexin V- PI apoptosis flow cytometry; J: EdU). Abbreviations: MC, macrophage culture medium; HMC, H. pylori-infected macrophage culture medium; IF, Immunofluorescence. The data are presented as the mean ± S.D. after triplicate Two groups were compared by t-test, multiple groups were compared by one-way analysis of variance (ANOVA). Note: *P < 0.05, **P < 0.01

Article Snippet: Human CCL3 ELISA kit was purchased from CUSABIO (CSB-E04662h, China).

Techniques: Expressing, Staining, Disruption, Flow Cytometry, Infection, Immunofluorescence

Journal: Cell communication and signaling : CCS

Article Title: Helicobacter pylori disrupts gastric mucosal homeostasis by stimulating macrophages to secrete CCL3.

doi: 10.1186/s12964-024-01627-5

Figure Lengend Snippet: Fig. 7 H. pylori-infected macrophage culture medium and chemokine CCL3 disrupt gastric mucosal barrier through P38 phosphorylation. A The P38 phosphorylation level in GES-1 treated with different conditioned media. B IF detected P38 phosphorylation levels in GES-1 treated with different conditioned media. DAPI was used for nuclear staining (blue), P-P38 stained in red. C, D The P38 phosphorylation level in GES-1 cells with recombinant CCL3 protein (160 ng/mL) and CCL3 overexpressing macrophage medium. E, F The P38 phosphorylation level in GES-1 cells treated with CCL3 neutralizing antibody (100 ng/mL) and CCL3 receptor inhibitor (Maraviroc, 100 nM). G Effects of P38 phosphorylation inhibitor (SB203580, 10 μM) on TEER of MKN28 cells. H IF detected the effect of P38 phosphorylation inhibitor on tight junctions between MKN28 cells. DAPI was used for nuclear staining (blue), Occludin was used for tight junction staining (green), arrows showed the disruption of tight junctions between cells. I, J The protein expression level of tight junction in GES-1 cells treated with P38 phosphorylation inhibitor. K, L Apoptosis and proliferation of GES-1 treated with P38 phosphorylation inhibitor (K: Annexin V- PI apoptosis flow cytometry; L: EdU). Abbreviations: NC, normal culture medium; MC, macrophage culture medium; HMC, H. pylori-infected macrophage culture medium; PV, pcDNA vector; IF, Immunofluorescence. The data are presented as the mean ± S.D. after triplicate. Two groups were compared by t-test, multiple groups were compared by one-way analysis of variance (ANOVA). Note: *P < 0.05, **P < 0.01

Article Snippet: Human CCL3 ELISA kit was purchased from CUSABIO (CSB-E04662h, China).

Techniques: Infection, Phospho-proteomics, Staining, Recombinant, Disruption, Expressing, Flow Cytometry, Plasmid Preparation, Immunofluorescence

Journal: Cell communication and signaling : CCS

Article Title: Helicobacter pylori disrupts gastric mucosal homeostasis by stimulating macrophages to secrete CCL3.

doi: 10.1186/s12964-024-01627-5

Figure Lengend Snippet: Fig. 9 Schematic diagram of the mechanism of gastric mucosal damage caused by CCL3 which is secreted by H. pylori infected macrophages. H. pylori induces the expression of the chemokine CCL3 by macrophages via the JAK1-STAT1 pathway, and CCL3 disrupts the gastric mucosal barrier via P38 phosphorylation

Article Snippet: Human CCL3 ELISA kit was purchased from CUSABIO (CSB-E04662h, China).

Techniques: Infection, Expressing, Phospho-proteomics

Journal: Frontiers in Immunology

Article Title: Targeting Pin1 to overcome immunosuppressive tumor microenvironment in MSS colorectal cancer

doi: 10.3389/fimmu.2025.1677029

Figure Lengend Snippet: Pin1 promotes Tregs chemotactic migration through the CCL3-CCR5 axis. (A) Correlation analysis of Pin1 with CCL chemokines expression. (B, C) qRT-PCR and ELISA assessing CCL3 expression in SW480 and HT29 cells after Pin1 knockdown. (D) Schematic representation of PBMCs isolated from human peripheral blood; Treg cells sorted by flow cytometry, these populations co-cultured with siNC- or siPin1-CRC cells, respectively. (E, F) Level of Treg markers (CD25 + CD127 - ) on PBMCs in the co-culture system detected by flow cytometry. (G) Effect of supernatants from Pin1-knockdown cells treated with recombinant CCL3 protein on Treg chemotactic capacity. (H) Correlation between CCL3 and CCR5 expression in CRC tissues. (I) CCR5-neutralizing antibodies abolish differences in Treg chemotaxis between the siNC and siPin1 groups.

Article Snippet: The concentrations of CCL3 in culture media were evaluated using Human CCL3 (MIP-1a) ELISA Kit (liankebio, cat.no.

Techniques: Migration, Expressing, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Knockdown, Isolation, Flow Cytometry, Cell Culture, Co-Culture Assay, Recombinant, Chemotaxis Assay

Journal: Frontiers in Immunology

Article Title: Targeting Pin1 to overcome immunosuppressive tumor microenvironment in MSS colorectal cancer

doi: 10.3389/fimmu.2025.1677029

Figure Lengend Snippet: Pin1 regulates CCL3 expression to promote Treg recruitment and CAFs activation through the NF-κB signaling pathway. (A) Validation of Pin1 interaction with p65 using CO-IP. (B) Co-localization of Pin1 with P65 confirmed using IF. (C) The main components of the NF-κB signaling pathway on knockdown of Pin1, as determined using WB. (D) Representative images of p65 IHC staining in subcutaneous tumors in mice (N.S. + IgG, Pin1 + IgG) (n=5 per group). (E) qRT-PCR assessed CCL3 expression in SW620 cells after Pin1 overexpression with/without NF-κB inhibitor treatment. (F) NF-κB inhibitor eliminated Pin1-overexpression-induced differences in Treg chemotaxis. (G) NF-κB inhibitor abrogated the activating effect of Pin1 overexpression on CAFs, as analyzed via IF.

Article Snippet: The concentrations of CCL3 in culture media were evaluated using Human CCL3 (MIP-1a) ELISA Kit (liankebio, cat.no.

Techniques: Expressing, Activation Assay, Biomarker Discovery, Co-Immunoprecipitation Assay, Knockdown, Immunohistochemistry, Quantitative RT-PCR, Over Expression, Chemotaxis Assay

Journal: Heliyon

Article Title: Salivary and serum nitric oxide synthase, macrophage inflammatory protein 1 alpha and macrophage migration inhibitory factor levels in periodontal disease

doi: 10.1016/j.heliyon.2024.e25888

Figure Lengend Snippet: Mean MIP-1α concentrations in serum and saliva.

Article Snippet: Commercial ELISA kits were purchased to determine MIF (human MIF ELISA kit, Wuhan USCN Business Corporation, Lot: L221207992, USA), MIP-1α (human MIP-1α ELISA kit, Wuhan USCN Business Corporation, Lot: L221207980, USA), and iNOS (human NOS 2 ELISA kit, Wuhan USCN Business Corporation, Lot: L221207994, USA) levels in serum and saliva.

Techniques:

Journal: Heliyon

Article Title: Salivary and serum nitric oxide synthase, macrophage inflammatory protein 1 alpha and macrophage migration inhibitory factor levels in periodontal disease

doi: 10.1016/j.heliyon.2024.e25888

Figure Lengend Snippet: Shows macrophage inflammatory protein 1 alpha (MIP-1α) concentrations in serum and saliva of patients with periodontally healthy (PH), gingivitis (G), stage I–II periodontitis (P1) and stage III–IV periodontitis (P2). Data are expressed as mean ± standard error. Differences were considered significant when p value was less than 0.05. *p = 0,006; **p < 0,05; # p = 0,016.

Article Snippet: Commercial ELISA kits were purchased to determine MIF (human MIF ELISA kit, Wuhan USCN Business Corporation, Lot: L221207992, USA), MIP-1α (human MIP-1α ELISA kit, Wuhan USCN Business Corporation, Lot: L221207980, USA), and iNOS (human NOS 2 ELISA kit, Wuhan USCN Business Corporation, Lot: L221207994, USA) levels in serum and saliva.

Techniques:

Journal: Journal of Inflammation Research

Article Title: Alpha-Momorcharin Inhibits Proinflammatory Cytokine Expression by M1 Macrophages but Not Anti-Inflammatory Cytokine Expression by M2 Macrophages

doi: 10.2147/JIR.S372306

Figure Lengend Snippet: Inflammatory cytokines and anti-inflammatory cytokines expression in M0, M1 and M2 macrophages. ( A ) TNF- α; ( B ) IL-1β; ( C ) IL-6; ( D ) IL-8; ( E ) MCP-1; ( F ) MIP-1α; ( G ) IL-RA; ( H ) IL-10; ( I ) TGF-β1; ( J ) EGF; ( K ) VEGF-A; ( L ) CCL22. The Orange-red columns are the M0-type macrophage groups, which was not treated with LPS; the blue columns are the M1-type macrophage groups; the purple columns are the M2-type macrophage groups. These macrophages were treated with 0.3 μg/mL α-MMC for 0 hours, 2 hours, 8 hours, 24 hours, or 48 hours. The data shown are individual values with the mean ± SEM; n = 5. * P < 0.05 significantly different from the M0 macrophage group of 0 hours; # P < 0.05 significantly different from the M1 macrophage group of 0 hours; n.s. No significant difference from the control group. One-way analysis of variance, Tukey’s multiple comparison tests.

Article Snippet: The following ELISA kits produced by Immunoway Co., Ltd. were used: Human TNF-α ELISA Kit (KE1372), Human IL-1β ELISA Kit (KE1379), Human IL-6 ELISA Kit (KE1368), Human IL-8 ELISA Kit (KE1360), Human MIP-1α ELISA Kit (KE 1052), Human MCP-1 ELISA Kit (KE1511), Human IL-1RA ELISA Kit (KE1030), Human IL-10 ELISA Kit (KE1369), Human EGF ELISA Kit (KE1012), Human VEGF-A ELISA Kit (KE1374), Human TGF-β ELISA Kit (KE1373), and Human CCL22 ELISA Kit (KE1203).

Techniques: Expressing

Journal: Journal of Inflammation Research

Article Title: Alpha-Momorcharin Inhibits Proinflammatory Cytokine Expression by M1 Macrophages but Not Anti-Inflammatory Cytokine Expression by M2 Macrophages

doi: 10.2147/JIR.S372306

Figure Lengend Snippet: The expression levels of the inflammatory cytokines TNF-α, IL-1β, IL-6, MIP-1α and MCP-1 in lung tissues and plasma. ( A – E ) TNF-α, IL-1β, IL-6, MIP-1α and MCP-1 levels in lung tissues; ( F – J ) TNF-α, IL-1β, IL-6, MIP-1α and MCP-1 levels in plasma. The data shown are individual values with the mean ± SEM; n = 8. * P < 0.05 significantly different from the sham group; # P < 0.05 significantly different from the LPS-induced model group; § P < 0.05 significantly different from the control group; n.s. No significant difference from the control group. One-way analysis of variance, Tukey’s multiple comparison tests.

Article Snippet: The following ELISA kits produced by Immunoway Co., Ltd. were used: Human TNF-α ELISA Kit (KE1372), Human IL-1β ELISA Kit (KE1379), Human IL-6 ELISA Kit (KE1368), Human IL-8 ELISA Kit (KE1360), Human MIP-1α ELISA Kit (KE 1052), Human MCP-1 ELISA Kit (KE1511), Human IL-1RA ELISA Kit (KE1030), Human IL-10 ELISA Kit (KE1369), Human EGF ELISA Kit (KE1012), Human VEGF-A ELISA Kit (KE1374), Human TGF-β ELISA Kit (KE1373), and Human CCL22 ELISA Kit (KE1203).

Techniques: Expressing

Journal: International Journal of Molecular Sciences

Article Title: Circulating Clues in Ménière’s Disease: Elevated Cell-Free DNA and a Pro-Inflammatory Signature in Patients’ Blood

doi: 10.3390/ijms27041948

Figure Lengend Snippet: Pro-inflammatory cytokines and chemokines increased in plasma from patients with Ménière’s disease (MD). Plasma concentrations of ( A ) IL-1β, ( B ) CCL3/MIP-1α, and ( C ) CCL27 were measured in healthy controls (HCs) and patients with MD. Data are presented as mean ± SD; n = 4. * p < 0.05. Individual experiments were performed in triplicate.

Article Snippet: Human IL-1β and MIP-1α/CCL3 were measured using the Human IL-1β ColorStep ELISA Kit (Assay Genie, Dublin, Ireland, cat# AEFI01732) and Human MIP-1α (CCL3) ColorStep ELISA Kit (Assay Genie, cat# AEFI03335), respectively.

Techniques: Clinical Proteomics