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: BMC medicine

Article Title: Plasma CCL3 predicts adverse heart failure outcomes in patients with arrhythmogenic cardiomyopathy.

doi: 10.1186/s12916-025-04024-y

Figure Lengend Snippet: Fig. 3 Macrophage-mediated inflammation in myocardium near the fibro-fatty tissues of ACM patients. A The mRNA expression of CCL3 in RV myocardium from ACM patients and NC; (B) The IHC staining of CCL3 in RV myocardium from ACM patients and NC; (C) Opal multicolor IHC staining of CCL3, CD68, and DAPI in RV myocardium from ACM patients and NC; (D) The correlation between the mRNA level of CCL3 in RV myocardium and the level of saturated TGs in RV fibro-fatty tissue from ACM patients; (E) The correlation between the mRNA level of CCL3 and duration from initial symptom to HTx; (F) The correlation between the mRNA level of CCL3 and duration from heart failure to HTx. RV, right ventricle; ACM, arrhythmogenic cardiomyopathy; NC, normal control; HTx, heart transplantation

Article Snippet: The concentrations of CCL3 in plasma were quantified using a commercial kit (Proteintech, KE00092).

Techniques: Expressing, Immunohistochemistry, Control, Transplantation Assay

Journal: bioRxiv

Article Title: Differential effects of voclosporin and tacrolimus on insulin secretion from human islets

doi: 10.1101/2020.04.30.071324

Figure Lengend Snippet: Insulin protein measured by radioimmunoassay from 20 islets treated for 48 hours with VCS or TAC (concentrations as indicate) after acid-ethanol extraction of insulin.

Article Snippet: Samples were stored at −20 °C and insulin secretion was quantified using human insulin radioimmunoassay kits (Cedarlane).

Techniques: RIA Assay, Extraction

Journal: iScience

Article Title: Blocking CCL3-mediated neutrophil recruitment into the brain alleviates immunopathology following severe enterovirus 71 infection

doi: 10.1016/j.isci.2024.111388

Figure Lengend Snippet: CCL3 and CCL2 showed the most robust increases in CSF of patients and mice during severe EV71 infection (A) Density distribution of 27 cytokines fold change of control (red), mild (blue) or critical (green)/control, in the sera of EV71-infected patients. (B) QPCR analysis of cytokines fold changes of critical/recovered CSF of patients. (C and D) CCL3 (C) and CCL2 (D) protein levels in CSF of patients with EV71 and brainstem encephalitis (severe immunopathology) at either the acute phase or recovery phase, measured by ELISA. (E and F) CCL3 (E) and CCL2 (F) protein levels in brain tissues of EV71-infected mice, measured by ELISA. Data are representative of three independent experiments. Data are reported as the mean ± SD, n = 10. ∗ p < 0.05 and ∗∗ p < 0.01 vs isotype antibody or WT mice (unpaired, two-tailed Student’s t test).

Article Snippet: human CCL3 ELISA kit , Boster Biological Engineering Co.,Ltd , EK0448.

Techniques: Infection, Control, Enzyme-linked Immunosorbent Assay, Two Tailed Test

Journal: iScience

Article Title: Blocking CCL3-mediated neutrophil recruitment into the brain alleviates immunopathology following severe enterovirus 71 infection

doi: 10.1016/j.isci.2024.111388

Figure Lengend Snippet: The deficiency of CCL3 reduces neutrophils infiltration in brain and mortality of severe EV71-infected mice 2 weeks old WT, Ccl3 −/− and Ccl2 −/− C57BL/6 mice were injected i.p. with 100LD50/100 μL/mouse of EV71. (A and B) Leukocytes were isolated from brain tissues by Ficoll Reagent and stained with anti-CD45-FITC and anti-Ly6G-PE on day 6 dpi. The percentages of neutrophils (CD45 + Ly6G + ) in total leukocytes were detected by FCM assay (A) and the corresponding statistical analyses are shown (B). (C and D) Immunofluorescence histochemistry analysis Ly6G for neutrophils in brain tissues (C) and the mean fluorescence intensity (MFI) (D) on day 6 dpi. (E and F) HE staining (E) and pathological scores of brain tissues (F) on day 6 dpi. (G–I) The mortality (G), body weight (H) and clinical scores (I) of mice (A) were monitored and recorded daily after infection. Data are representative of three independent experiments. Data are reported as the mean ± SD, n = 10. ∗ p < 0.05 and ∗∗ p < 0.01 vs isotype antibody or WT mice (unpaired, two-tailed Student’s t test).

Article Snippet: human CCL3 ELISA kit , Boster Biological Engineering Co.,Ltd , EK0448.

Techniques: Infection, Injection, Isolation, Staining, Immunofluorescence, Fluorescence, Two Tailed Test

Journal: iScience

Article Title: Blocking CCL3-mediated neutrophil recruitment into the brain alleviates immunopathology following severe enterovirus 71 infection

doi: 10.1016/j.isci.2024.111388

Figure Lengend Snippet: CCL3 recruit neutrophils into the brain through CCR1/3(human) and CCR5(mouse) (A) Transwell assay for chemotactic ability of neutrophils from EV71-infected mice or PBS-injected mice, in which the CCL3 and CCL2 proteins were placed in the lower chamber and the neutrophils were seeded in the upper chamber and 3 h later the neutrophils that had transversed into the lower chamber were counted. Positive control: fMLP; negative control: RPMI-160 medium. (B) qPCR analysis chemokine receptors expression in neutrophils from control, mild and critical EV71 infection patients. (C and E) Expression of CCR1 (C), CCR3 (D) and CCR5 (E) on neutrophils in human CSF from patients with EV71 infection during either the acute phase or recovery phase, measured by qRT-PCR. (F–H) Expression of Ccr1 (F), Ccr3 (G) and Ccr5 (H) on neutrophils in brain tissues from EV71-infected mice or PBS-injected negative control mice, measured by qRT-PCR.

Article Snippet: human CCL3 ELISA kit , Boster Biological Engineering Co.,Ltd , EK0448.

Techniques: Transwell Assay, Infection, Injection, Positive Control, Negative Control, Expressing, Control, Quantitative RT-PCR

Journal: iScience

Article Title: Blocking CCL3-mediated neutrophil recruitment into the brain alleviates immunopathology following severe enterovirus 71 infection

doi: 10.1016/j.isci.2024.111388

Figure Lengend Snippet: Inhibition of CCL3 reduces the neutrophils infiltration in brains and the mortality of EV71 infected mice (A–F) EV71-infected mice (100LD 50 /100 μL/mouse of EV71) were injected i.p. with isotype or anti-CCL3 antibody (100 μg/100 μL/mouse). (A) ELISA analysis the CCL3 expression in mice brain tissues. (C and B) Leukocytes were isolated from brain tissues of EV71-infected mice by use of the Ficoll Reagent on day 6 dpi and stained with anti-CD45-FITC and anti-Ly6G-PE. The percentages of neutrophils (CD45+Ly6G+) in total leukocytes were detected by FCM assay (C) and the corresponding statistical analyses are shown (B). (D–F) The mortality (D), body weight (E) and clinical scores (F) of mice were monitored and recorded daily after infection. (G–L) EV71-infected mice (100LD 50 /100 μL/mouse of EV71) were injected i.p. with Mixed-scrables or Mixed-mmu-mirs (90 μg/g body weight), (G) qPCR analysis the Ccl3 mRNA expression in mice brain tissues. (H and I) Leukocytes were isolated from brain tissues of EV71-infected mice by use of the Ficol Reagent on day 6 dpi and stained with anti-CD45-FITC and anti-Ly6G-PE. The percentages of neutrophils (CD45 + Ly6G + ) in total leukocytes were detected by FCM assay (H) and the corresponding statistical analyses are shown (I). (J–L) The mortality (J), body weight (K) and clinical scores (L) of mice (G) were monitored and recorded daily after infection.

Article Snippet: human CCL3 ELISA kit , Boster Biological Engineering Co.,Ltd , EK0448.

Techniques: Inhibition, Infection, Injection, Enzyme-linked Immunosorbent Assay, Expressing, Isolation, Staining

Journal: iScience

Article Title: Blocking CCL3-mediated neutrophil recruitment into the brain alleviates immunopathology following severe enterovirus 71 infection

doi: 10.1016/j.isci.2024.111388

Figure Lengend Snippet:

Article Snippet: human CCL3 ELISA kit , Boster Biological Engineering Co.,Ltd , EK0448.

Techniques: Virus, Recombinant, Enzyme-linked Immunosorbent Assay, Isolation, Knock-Out, Software

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