ccl2 neutralizing antibody cat#be0185  (Bio X Cell)

Journal: Journal of Neuroinflammation

Article Title: Sigma-1 receptor regulates HIV-1 and methamphetamine-induced endothelial/pericyte barrier impairment via strain-specific inflammatory responses and mitochondrial dysregulation

doi: 10.1186/s12974-026-03750-1

Figure Lengend Snippet: METH enhances HIV-1 NL4-3 replication in HBVPs independently of Sigma-1R. A - B Impact of METH (25 µM) on HIV-1 replication as measured by p24 antigen release levels in HBVPs infected with HIV-1 NL4-3 ( A ) or JR-CSF ( B ). Data are means ± SEM ( n = 3). C - D Impact of METH (25 µM) on HIV-1 replication as measured by HIV-1 Gag mRNA expression levels in HBVPs infected with HIV-1 NL4-3 ( C ) or JR-CSF ( D ) ( n = 4–5). E Impact of pretreatment with S1RA (10 µM) for 6 h on NL4-3 HIV-1 replication in the presence and absence of METH ( n = 3–4). F Impact of HBVP pretreatment with the CXCR4 chemokine receptor antagonist AMD070 (5 µM) for 1 h on HIV-1 NL4-3 replication in the presence and absence of METH ( n = 4). G The heat map demonstrating the impact of HIV-1 NL4-3 infection and/or METH treatment for 72 h on gene expression profile of 42 ISGs in HBVPs ( n = 6). Genes with high expression levels are represented in shades of red, while those with low expression levels are shown in shades of green. Gene names are shown on the x axis. Red arrows indicate genes that were significantly differentially regulated in the HIV-1 NL4-3 + METH group compared to the control group, as determined by the RT² Profiler PCR Array ( p < 0.05). H - K RT-qPCR analysis of mRNA expression of CCL2 ( H ), MX2 ( I ), IFI30 ( J ), and PRKD2 ( K ) in HBVPs infected with HIV-1 NL4-3 and/or treated with METH ( n = 12). L Impact of blocking endogenous CCL2 with anti-human CCL2 neutralizing antibody on p24 release in HIV-1 NL4-3-infected HBVPs, with or without METH, at 72 h post-infection ( n = 6). M Impact of pretreatment with the CXCR4 chemokine receptor antagonist AMD070 (5 µM) for 1 h on CCL2 release in the presence and absence of METH at 72 h post-infection ( n = 6). Data are means ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001 and **** p < 0.0001. Abbreviations as in Fig. ; CCL2 - C-C motif chemokine ligand 2

Article Snippet: To neutralize CCL2, cultures were incubated with an anti-human CCL2 neutralizing antibody (8 μg/mL; R&D Systems, Cat# AF-279-NA) or with normal goat IgG as a control (R&D Systems, Cat# AB-108-C).

Techniques: Infection, Expressing, Gene Expression, Control, Quantitative RT-PCR, Blocking Assay

Journal: Journal of Neuroinflammation

Article Title: Sigma-1 receptor regulates HIV-1 and methamphetamine-induced endothelial/pericyte barrier impairment via strain-specific inflammatory responses and mitochondrial dysregulation

doi: 10.1186/s12974-026-03750-1

Figure Lengend Snippet: Synergistic impact of METH and CXCR4-Tropic HIV-1 on pericyte-dependent endothelial barrier breakdown via CXCR4/CCL2-driven viral replication and Sigma-1R-mediated mitochondrial and inflammatory dysregulation. This proposed model depicts intersecting pathways through which CXCR4-tropic HIV-1 and METH synergistically compromise endothelial barrier integrity via viral replication, Sigma-1R-mediated mitochondrial dysfunction, and modulation of IL6-associated inflammatory response. Notably, METH-enhanced replication of CXCR4-tropic HIV-1 in pericytes appears to occur independently of the Sigma-1R signaling ( www.BioRender.com )

Article Snippet: To neutralize CCL2, cultures were incubated with an anti-human CCL2 neutralizing antibody (8 μg/mL; R&D Systems, Cat# AF-279-NA) or with normal goat IgG as a control (R&D Systems, Cat# AB-108-C).

Techniques:

Journal: Poultry Science

Article Title: Prolactin promotes transepithelial migration of lymphocytes through CCL2

doi: 10.1016/j.psj.2025.105322

Figure Lengend Snippet: (A) The co-culture setup of mammary epithelial cells and T lymphocytes. (B) The number of lymphocytes in the lower chamber was counted by a cell counter, ** P < 0.01; (C) Lymphocytes across breast epithelial cells were observed by fluorescence confocal microscopy; the left and right images are the cross-section and a vertical view of Transwell chamber, respectively. (D) The number of migrated lymphocytes under prolactin stimulation with or without CCL2 neutralization was quantified using a cell counter. CCL2 neutralizing antibody was added to the lower chamber simultaneously with prolactin, ** P < 0.01; (E) Lymphocytes across breast epithelial cells were observed by fluorescence confocal microscopy; the left and right images are the cross-section and a vertical view of the Transwell chamber, respectively.

Article Snippet: One group was treated with 50 ng/mL prolactin (1445-PR, R&D Systems) and 5 μg/mL CCL2 neutralizing antibodies (goat anti-mouse CCL2, AF-479-NA, R&D Systems) to block CCL2 ( ).

Techniques: Co-Culture Assay, Fluorescence, Confocal Microscopy, Neutralization

Journal: Oncoimmunology

Article Title: Glutamate promotes CCL2 expression to recruit tumor-associated macrophages by restraining EZH2-mediated histone methylation in hepatocellular carcinoma

doi: 10.1080/2162402X.2025.2497172

Figure Lengend Snippet: Glutamate increases CCL2 expression on hepatoma cells.

Article Snippet: For the CCL2-CCR2 studies, mice were treated with CCL2 neutralizing antibody (2 mg/kg, Bio X cell, cat#BE0185) or CCL2 mAb in combination with MK801 intraperitoneally twice a week for 2 weeks.

Techniques: Expressing

Journal: Oncoimmunology

Article Title: Glutamate promotes CCL2 expression to recruit tumor-associated macrophages by restraining EZH2-mediated histone methylation in hepatocellular carcinoma

doi: 10.1080/2162402X.2025.2497172

Figure Lengend Snippet: EZH2 pathway is essential for glutamate-induced CCL2 expression in HCC.

Article Snippet: For the CCL2-CCR2 studies, mice were treated with CCL2 neutralizing antibody (2 mg/kg, Bio X cell, cat#BE0185) or CCL2 mAb in combination with MK801 intraperitoneally twice a week for 2 weeks.

Techniques: Expressing

Journal: Oncoimmunology

Article Title: Glutamate promotes CCL2 expression to recruit tumor-associated macrophages by restraining EZH2-mediated histone methylation in hepatocellular carcinoma

doi: 10.1080/2162402X.2025.2497172

Figure Lengend Snippet: EZH2-mediated H3K27me3 on the CCL2 promoter to control CCL2 expression.

Article Snippet: For the CCL2-CCR2 studies, mice were treated with CCL2 neutralizing antibody (2 mg/kg, Bio X cell, cat#BE0185) or CCL2 mAb in combination with MK801 intraperitoneally twice a week for 2 weeks.

Techniques: Control, Expressing

Journal: MedComm

Article Title: PDPN+ cancer‐associated fibroblasts enhance gastric cancer angiogenesis via AKT/NF‐κB activation and the CCL2‐ACKR1 axis

doi: 10.1002/mco2.70037

Figure Lengend Snippet: High expression of podoplanin (PDPN), CD31 and chemokine (CC‐motif) ligand 2 (CCL2) in gastric cancer (GC) is associated with poor prognosis. (A) Representative hematoxylin and eosin (H&E) and immunohistochemical images showing PDPN, CD31, and CCL2 protein expression in GC and normal gastric mucus; (B) Semiquantitative analysis of PDPN, CD31, and CCL2 protein expression between GC and normal gastric mucosa; (C–H) Kaplan–Meier survival curves with log‐rank tests depicting disease‐free survival (C, E, G) and overall survival (D, F, H) of GC patients ( n = 400) based on the expression levels of PDPN, CCL2, and intersection of all three proteins in cancer tissues in the in‐house cohort.

Article Snippet: Other antibodies include CD31 (Cat# ab81289, Abcam), CCL2 neutralizing antibody (Cat# MAB679, R&D), human CCL2 (Cat# 300‐04‐5, PeproTech).

Techniques: Expressing, Immunohistochemical staining

Journal: MedComm

Article Title: PDPN+ cancer‐associated fibroblasts enhance gastric cancer angiogenesis via AKT/NF‐κB activation and the CCL2‐ACKR1 axis

doi: 10.1002/mco2.70037

Figure Lengend Snippet: Podoplanin (PDPN)(+) cancer‐associated fibroblasts (CAFs) induce angiogenesis by secreting chemokine (CC‐motif) ligand 2 (CCL2). (A) Cytokine profiles produced by conditioned medium (CM) of PDPN(+) CAFs and PDPN(−) CAFs were examined by RayBio Human Cytokine Antibody Array. The red, blue, and purple frame represents the most significantly overexpressed cytokines. (B) Top 10 cytokines significantly upregulated in CM of PDPN(+) CAFs compared to PDPN(−) CAFs (*** p < 0.001). (C) Enzyme‐linked immunosorbent assay (ELISA) results showing the amount of soluble CCL2 produced by PDPN(+) CAFs, PDPN(−) CAFs, normal fibroblasts (NFs), and human umbilical vein endothelial cells (HUVECs). (D) Uniform manifold approximation and projection (UMAP) of endothelial cells representing three unique cell states, color‐coded by their corresponding cell lineage or subtype. Each dot in the UMAP represents a single cell. (E) Circle plots showing the cellular interactions of CAFs and endothelial cells (ECs) involved in the CCL signaling pathway network in gastric cancer (GC). CAFs and ECs were the core of the cellular interaction network (edge width represents the numbers of interactions and node size represents the abundance of cell populations). (F, G) Wound healing assay (F) and tube formation assay (G) on HUVECs cultured in CM of PDPN(+) CAFs with or without a neutralizing antibody against CCL2 (anti‐CCL2). Scale bar = 100 µm; ** p < 0.01. (H, I) Wound healing assay (H) and tube formation assay (I) on HUVECs treated with or without CCL2 (scale bar = 100 µm; ** p < 0.01). (J) Representative immunofluorescent staining for PDPN, CCL2, and CD31 in clinical GC sample (scale bar = 40 µm). (K) Immunoblot analysis of p‐PI3K, PI3K, p‐AKT, AKT expression in HUVECs treated with CM from PDPN(+) CAFs and CCL2 neutralizing antibody (anti‐CCL2). (L) Immunoblot analysis of p‐PI3K, PI3K, p‐AKT, AKT expression in HUVECs treated with CCL2. (M) Violin plots showing the expression of genes involved in the CCL signaling pathway. CCL2 and CCL11 are primarily expressed in fibroblast subsets, while their potential receptor ACKR1, is predominantly expressed in endothelial cells. (N) Immunoblot analysis of p‐PI3K, PI3K, p‐AKT, AKT expression in HUVECs with indicated treatment.

Article Snippet: Other antibodies include CD31 (Cat# ab81289, Abcam), CCL2 neutralizing antibody (Cat# MAB679, R&D), human CCL2 (Cat# 300‐04‐5, PeproTech).

Techniques: Produced, Ab Array, Enzyme-linked Immunosorbent Assay, Wound Healing Assay, Tube Formation Assay, Cell Culture, Staining, Western Blot, Expressing

Journal: MedComm

Article Title: PDPN+ cancer‐associated fibroblasts enhance gastric cancer angiogenesis via AKT/NF‐κB activation and the CCL2‐ACKR1 axis

doi: 10.1002/mco2.70037

Figure Lengend Snippet: Podoplanin (PDPN) regulates NF‐κB activity in cancer‐associated fibroblasts (CAFs) through AKT/IKK signaling, leading to chemokine (CC‐motif) ligand 2 (CCL2) secretion. (A) Heatmap representing significantly dysregulated genes from RNA‐seq analysis of PDPN(+) CAFs and PDPN(−) CAFs ( n = 3). (B) Pathway analysis of differentially expressed genes enriched in PDPN(+) CAFs compared to PDPN(−) CAFs. (C) Immunoblot analysis of nuclear and cytoplasmic p‐P65 and P65 protein expression in PDPN(+) CAFs, PDPN(−) CAFs and normal fibroblasts (NFs). (D) Representative immunofluorescent staining of p‐P65 in PDPN(+) CAFs, PDPN(−) CAFs, and NFs (scale bar = 25 µm). (E) Quantitative reverse transcription polymerase chain reaction (qRT‐PCR) results showing the differential mRNA expression levels of CCL2 in PDPN(+) CAFs transfected with P65 siRNA or control siRNA ( n = 3; ** p < 0.01). (F) Enzyme‐linked immunosorbent assay (ELISA) results showing the amount of soluble CCL2 in conditioned medium (CM) from PDPN(+) CAFs transfected with P65 siRNA or control siRNA (** p < 0.01). (G, H) Wound healing assay (G) and tube formation assay (H) on human umbilical vein endothelial cells (HUVECs) treated with CM from PDPN(+) CAFs transfected with P65 siRNA or control siRNA (scale bar = 100 µm; ** p < 0.01). (I) Chromatin immunoprecipitation (ChIP) assay was performed to verify P65 binding to the CCL2 promoter. CCL2 promoter segments were quantified using qRT‐PCR, with results normalized against IgG. Data are presented as the mean ± SD from three independent experiments is presented (*** p < 0.001). (J) Top: Schematic representation of the CCL2 reporter construct. The consensus P65 binding sequences are marked as with blue box. The consensus sequence and the putative P65 binding site sequences are shown. Bottom: Effects of ectopic expression of P65 siRNA on wild type (WT) and mutant (MUT) CCL2 promoter reporter activity (** p < 0.01). (K) Effects of ectopic expression of P65 siRNA (siP65#1), AKT inhibitors perifosine (10 µmol/L), NF‐κB inhibitor pyrrolidine dithiocarbamate (PDTC, 0.05 µmol/L) on CCL2 promoter reporter activity (** p < 0.01). (L) Immunoblot analysis of AKT/IKK pathway in PDPN(+) CAFs, PDPN(−) CAFs and NFs. (M) Immunoblot analysis of AKT/IKK/NF‐κB pathway and nuclear/cytoplasmic distribution of P65 in PDPN(+) CAFs treated with or without perifosine (10 µmol/L) and LY294002 (20 µmol/L) for 24 h.

Article Snippet: Other antibodies include CD31 (Cat# ab81289, Abcam), CCL2 neutralizing antibody (Cat# MAB679, R&D), human CCL2 (Cat# 300‐04‐5, PeproTech).

Techniques: Activity Assay, RNA Sequencing, Western Blot, Expressing, Staining, Reverse Transcription, Polymerase Chain Reaction, Quantitative RT-PCR, Transfection, Control, Enzyme-linked Immunosorbent Assay, Wound Healing Assay, Tube Formation Assay, Chromatin Immunoprecipitation, Binding Assay, Construct, Sequencing, Mutagenesis

Journal: MedComm

Article Title: PDPN+ cancer‐associated fibroblasts enhance gastric cancer angiogenesis via AKT/NF‐κB activation and the CCL2‐ACKR1 axis

doi: 10.1002/mco2.70037

Figure Lengend Snippet: Targeting podoplanin (PDPN)(+) cancer‐associated fibroblasts (CAFs) restrains angiogenesis via inhibiting AKT/NF‐κB signaling. (A) Schematic diagram of in vivo animal experiment. (B) Tumor image (left) and tumor growth curve (right) of xenografted mice inoculated with HGC27 cells and PDPN(+) CAFs pretreated with or without perifosine (10 µmol/L) and Bay117082 (10 µmol/L; *** p < 0.001). (C) Representative images of hematoxylin and eosin (H&E) and immunofluorescent staining for CD31 as indicated in the xenografted tumors (scale bar = 40 µm; * p < 0.05). (D) Schematic view of the proposed mechanism. A schematic diagram illustrating the proposed mechanism by which PDPN(+) CAFs‐derived chemokine (CC‐motif) ligand 2 (CCL2) promotes angiogenesis in gastric cancer (GC). The Akt/NF‐κB pathway was significantly activated in PDPN(+) CAFs. P65 directly bind to the CCL2 promoter, thereby increasing the CCL2 transcription and secretion in CAFs; CCL2, which was transcriptional activated by P65 in PDPN(+) CAFs, sustains tumor angiogenesis by interacting with ACKR1 and activating PI3K/AKT signaling in endothelial cells. The scheme was drawn by biorender ( https://biorender.com/ ).

Article Snippet: Other antibodies include CD31 (Cat# ab81289, Abcam), CCL2 neutralizing antibody (Cat# MAB679, R&D), human CCL2 (Cat# 300‐04‐5, PeproTech).

Techniques: In Vivo, Staining, Derivative Assay