human ccl28 protein  (Cell Signaling Technology Inc)

Journal: bioRxiv

Article Title: IgE-producing cells on the move: CCR2 is a key regulator of IgE + plasma cell migration

doi: 10.64898/2025.12.18.695109

Figure Lengend Snippet: (A) Representative histograms showing surface CCR10 expression on IgE + (red) and IgG1 + (blue) gated cells; the filled grey histogram shows the isotype control staining. The accompanying graph shows CCR10 MFI on the gated IgE + and IgG1 + cells. (B) Representative plots of CCR10 expression on IgE + and IgG1 + GC-like B cells, PBs, and PCs. CCR10 MFI across the different IgE + and IgG1 + cell populations from several experiments. (C) The dot plots show the frequency of IgE + and IgG1 + cells after 3h of migration in response to RPMI only or two different concentrations of CCL28, 25nM (300 ng/mL) and 125nM (1.5 μg/mL). (D) CCL28 induced migration shown as a percentage of the IgE + and IgG1 + cells migrating in response to RPMI control. (E) The migration capacity of IgE + and IgG1 + GC-like B cells, plamablasts, and PC in response to CCL28. (F) CCL28 induced migration shown as a percentage of the cells migrating in response to RPMI control. Data are mean + s.d. Statistical analysis was performed using one-way ANOVA with Tukey’s multiple comparison test (B, D, F) or paired two-tailed t -test with Welch’s correction (A); *p< 0.05; **p< 0.01.

Article Snippet: More specifically, we tested the migration of IgE + and IgG1 + cells in response to recombinant human CXCL12 (R&D systems; 300ng/mL), CCL2 (Biolegend; 10ng/mL, 100ng/mL and 300ng/mL) and CCL28 (R&D systems; 300ng/mL and 1.5ug/mL).

Techniques: Expressing, Control, Staining, Migration, Comparison, Two Tailed Test

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Pericytes recruited by CCL28 promote vascular normalization after anti-angiogenesis therapy through RA/RXRA/ANGPT1 pathway in lung adenocarcinoma

doi: 10.1186/s13046-024-03135-3

Figure Lengend Snippet: CCL28 expression is upregulated after anti-angiogenesis therapy by hypoxia-sensitive transcription factor CEBPB in lung adenocarcinoma

Article Snippet: Briefly, pericytes treated with or without recombinant human CCL28 or CCR3 (R&D Systems, MAB155-100) neutralizing antibodies were collected and fixed by adding a cross-linking agent, formaldehyde, to stabilize the interactions between chromatin proteins and DNA.

Techniques: Expressing

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Pericytes recruited by CCL28 promote vascular normalization after anti-angiogenesis therapy through RA/RXRA/ANGPT1 pathway in lung adenocarcinoma

doi: 10.1186/s13046-024-03135-3

Figure Lengend Snippet: Tumor-derived CCL28 recruits pericytes to promote vascular normalization in the tumor microenvironment

Article Snippet: Briefly, pericytes treated with or without recombinant human CCL28 or CCR3 (R&D Systems, MAB155-100) neutralizing antibodies were collected and fixed by adding a cross-linking agent, formaldehyde, to stabilize the interactions between chromatin proteins and DNA.

Techniques: Derivative Assay

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Pericytes recruited by CCL28 promote vascular normalization after anti-angiogenesis therapy through RA/RXRA/ANGPT1 pathway in lung adenocarcinoma

doi: 10.1186/s13046-024-03135-3

Figure Lengend Snippet: Tumor-derived CCL28 promotes the expression of angiopoietin-1 via CCR3 in pericytes

Article Snippet: Briefly, pericytes treated with or without recombinant human CCL28 or CCR3 (R&D Systems, MAB155-100) neutralizing antibodies were collected and fixed by adding a cross-linking agent, formaldehyde, to stabilize the interactions between chromatin proteins and DNA.

Techniques: Derivative Assay, Expressing

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Pericytes recruited by CCL28 promote vascular normalization after anti-angiogenesis therapy through RA/RXRA/ANGPT1 pathway in lung adenocarcinoma

doi: 10.1186/s13046-024-03135-3

Figure Lengend Snippet: Retinoic acid signaling is activated by CCL28 in pericytes through CCR3 A , Volcano plot of changes in metabolic pathways after CCL28 stimulation. B Volcano plot of the enrichment of gene expression after CCL28 stimulation. C Diagram of the metabolic conversion process in the retinoic acid metabolic signaling pathway. D and E Expression of RDH13 and DHRS11 detected by qPCR and western blot with or without exogenous supplement of CCL28. F Correlation of expression of CCL28 with RDH13 in lung adenocarcinoma. G The protein level of DHRS11 and RDH13 stimulated with or without CCL28 and CCR3 neutralizing antibody in pericytes (left) and gray value was calculated(right). H Knockdown efficiency of RDH13 was confirmed by qPCR. I and J Relative expression of RXRα and ANGPT1 after knockdown of RDH13 with or without stimulation of CCL28. K Representative immunofluorescence images of PAN-CK, NG2, CCL28 with DHRS11 or RDH13 or Angiopoietin-1 on biopsy tissues from lung cancer patients (left panel). Scale bar = 100 μm. The correlation between the expression of CCL28 and the levels of DHRS11, RDH13, and angiopoietin-1 (right panel). Data with error bars are shown as mean ± SEM. Each symbol represents data from a replicate. Each panel is a representative experiment of at least three independent biological replicates. *, **, *** represent p < 0.05, p < 0.01 and p < 0.001, respectively. Abbreviation: MFI, Mean fluorescence intensity

Article Snippet: Briefly, pericytes treated with or without recombinant human CCL28 or CCR3 (R&D Systems, MAB155-100) neutralizing antibodies were collected and fixed by adding a cross-linking agent, formaldehyde, to stabilize the interactions between chromatin proteins and DNA.

Techniques: Gene Expression, Expressing, Western Blot, Knockdown, Immunofluorescence, Fluorescence

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Pericytes recruited by CCL28 promote vascular normalization after anti-angiogenesis therapy through RA/RXRA/ANGPT1 pathway in lung adenocarcinoma

doi: 10.1186/s13046-024-03135-3

Figure Lengend Snippet: Both CCL28 and retinoic acid could promote vascular normalization in vivo

Article Snippet: Briefly, pericytes treated with or without recombinant human CCL28 or CCR3 (R&D Systems, MAB155-100) neutralizing antibodies were collected and fixed by adding a cross-linking agent, formaldehyde, to stabilize the interactions between chromatin proteins and DNA.

Techniques: In Vivo

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Pericytes recruited by CCL28 promote vascular normalization after anti-angiogenesis therapy through RA/RXRA/ANGPT1 pathway in lung adenocarcinoma

doi: 10.1186/s13046-024-03135-3

Figure Lengend Snippet: CCL28 is involved in bevacizumab-mediated vascular normalization

Article Snippet: Briefly, pericytes treated with or without recombinant human CCL28 or CCR3 (R&D Systems, MAB155-100) neutralizing antibodies were collected and fixed by adding a cross-linking agent, formaldehyde, to stabilize the interactions between chromatin proteins and DNA.

Techniques:

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Pericytes recruited by CCL28 promote vascular normalization after anti-angiogenesis therapy through RA/RXRA/ANGPT1 pathway in lung adenocarcinoma

doi: 10.1186/s13046-024-03135-3

Figure Lengend Snippet: A schematic diagram of tumor microenvironment modulation effects of CCL28

Article Snippet: Briefly, pericytes treated with or without recombinant human CCL28 or CCR3 (R&D Systems, MAB155-100) neutralizing antibodies were collected and fixed by adding a cross-linking agent, formaldehyde, to stabilize the interactions between chromatin proteins and DNA.

Techniques:

Journal: Mucosal immunology

Article Title: Th22 cells are efficiently recruited in the gut by CCL28 as an alternative to CCL20 but do not compensate for the loss of Th17 cells in treated HIV-1-infected individuals.

doi: 10.1038/s41385-020-0286-6

Figure Lengend Snippet: Fig. 2 Expression of CCL28 by enterocytes in the duodenal mucosa. CCL28 production in the duodenal mucosa of treated HIV- 1-infected (n = 10) and uninfected individuals (n = 10). Chemokine expression was quantified per surface unit of epithelium using NIS- element (Nikon). CCL28 (red) was stained by immunohistochemistry on an ApoTome (Zeiss, original magnification ×63). Cell nuclei were counterstained (DAPI, blue). Representative treated HIV-1-infected and uninfected individuals are shown. Groups were compared with the Wilcoxon’s rank-sum test. Median bars are shown.

Article Snippet: In gradient experiments, CCL20 and CCL28 recombinant proteins (R&D Systems) were loaded in the bottom chamber with a CCL20/CCL28 ratio varying from 0:100% to 100:100% of their maximum chemotactic activity (10 and 200 ng/mL, respectively).

Techniques: Expressing, Infection, Staining, Immunohistochemistry

Journal: Mucosal immunology

Article Title: Th22 cells are efficiently recruited in the gut by CCL28 as an alternative to CCL20 but do not compensate for the loss of Th17 cells in treated HIV-1-infected individuals.

doi: 10.1038/s41385-020-0286-6

Figure Lengend Snippet: Fig. 3 Impact of the interactions between IL-17A/Th17 cells and IL-22/Th22 cells on CCL20 and CCL28 expression by enterocytes. Effect of IL-17A (white bars) and IL-22 (gray bars) on a CCL20 and b CCL28 mRNA expression by enterocytes. Monolayers of differentiated human primary enterocytes on transwell inserts were stimulated by 0.5, 5, and 50 ng/mL of cytokine. CCL20 and CCL28 mRNA was quantified in the enterocytes by qRT-PCR. CCL20 and CCL28 expression following cytokine stimulation was normalized relatively to the expression in unstimulated epithelial cells (set to 1) and expressed as fold change (log2 scale). Presented data were obtained from at least eight independent experiments performed with different donors. Cuzick’s test for trend was used to compare chemokine expression across the increasing concentrations of cytokines (the corresponding P value is shown on the line above the IL17-A and IL-22 bars); paired Wilcoxon’s test was used to compare chemokine expression upon stimulation vs. unstimulated condition (intradonor pairing); P value is shown above each bar; *P < 0.05; **P < 0.01. Means and SEM are shown. c Effect of Th22:IEC coculture on CCL20 and CCL28 mRNA expression by enterocytes. Enterocytes were cocultured with FACS-sorted Th22 or Th17 cells, added in the bottom chamber for 15 h. CCL20 and CCL28 mRNA were quantified in the enterocytes by qRT-PCR. CCL20 and CCL28 expression in Th22:IEC coculture was normalized relatively to the expression in Th17:IEC coculture (set to 1) and expressed as fold change (log2 scale). Enterocytes production of d CCL20 and e CCL28 proteins in Th22:IEC and Th17:IEC cocultures. CCL20 and CCL28 were quantified in the bottom chamber by ELISA. Presented data were obtained from ten independent experiments performed with different donors. Paired Wilcoxon’s test was used to compare CCL20 and CCL28 in Th22:IEC vs. Th17:IEC cocultures (intradonor pairing); *, P < 0.05. IEC, intestine epithelial cells. Means and SEM are shown.

Article Snippet: In gradient experiments, CCL20 and CCL28 recombinant proteins (R&D Systems) were loaded in the bottom chamber with a CCL20/CCL28 ratio varying from 0:100% to 100:100% of their maximum chemotactic activity (10 and 200 ng/mL, respectively).

Techniques: Expressing, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in medicine

Article Title: Activated Hepatic Stellate Cells Induce Infiltration and Formation of CD163 + Macrophages via CCL2/CCR2 Pathway.

doi: 10.3389/fmed.2021.627927

Figure Lengend Snippet: FIGURE 3 | The high level of CCL2 in aHSCs was associated with CD163+ macrophage infiltration and increased with liver fibrosis progression. (A) The primary aHSCs are typically fusiform and express the activation marker α-SMA, together with a high expression of CCL2 protein. (B) The aHSCs secrete high levels of CCL2 (Continued)

Article Snippet: When indicated, recombinant human CCL2 protein (rh CCL2, 2ng/ml, R&D Systems, Abingdon, UK) and INCB 3284 (100ng/ml, Tocris Bioscience, UK) were accordingly added, after which the macrophages were harvested, counted, and analyzed.

Techniques: Activation Assay, Marker, Expressing

Journal: Frontiers in medicine

Article Title: Activated Hepatic Stellate Cells Induce Infiltration and Formation of CD163 + Macrophages via CCL2/CCR2 Pathway.

doi: 10.3389/fmed.2021.627927

Figure Lengend Snippet: FIGURE 4 | CCL2 was responsible for macrophages infiltration and differentiation into M2 phenotype during liver fibrosis. (A) Representative images of macrophage infiltration under different chemotaxis treatments including aHSC, aHSC+INCB, Rh CCL2 and medium. (B) Statistical analysis of the number of macrophages (Continued)

Article Snippet: When indicated, recombinant human CCL2 protein (rh CCL2, 2ng/ml, R&D Systems, Abingdon, UK) and INCB 3284 (100ng/ml, Tocris Bioscience, UK) were accordingly added, after which the macrophages were harvested, counted, and analyzed.

Techniques: Chemotaxis Assay

Journal: Cell Death & Disease

Article Title: The chemokine receptor CCR10 promotes inflammation-driven hepatocarcinogenesis via PI3K/Akt pathway activation

doi: 10.1038/s41419-018-0267-9

Figure Lengend Snippet: CCR10-transfected HepG2 and LO2 cells were treated with either the CCR10 agonist-ligand CCL28 or the Akt inhibitor A6730. a Activation of the CCL28-CCR10 axis by CCL28 significantly increased Akt phosphorylation, PCNA protein expression, and b relative cell proliferation in both cell lines, while Akt inhibition produced the opposite effects. Relative cell proliferation is defined as the fold-change in proliferation relative to the untreated parent cell line. * P < 0.05 vs. CCR10 group. All values are reported as means ± standard errors of the mean (SEMs)

Article Snippet: For some in vitro experiments, HepG2 and LO2 cell lines were pre-treated with the pro-inflammatory cytokine TNF (concentrations as indicated; R&D Systems, Minneapolis, MN, USA) for 4 h, recombinant human CC chemokine ligand 28 (CCL28) (400 nM; R&D Systems) for 2 h, or the allosteric Akt inhibitor A6730 (10 μM; Sigma, St. Louis, MO, USA) for 2 h . Human CCL28 is a natural ligand-agonist for human CCR10 .

Techniques: Transfection, Activation Assay, Phospho-proteomics, Expressing, Inhibition, Produced

Journal: Cell Death & Disease

Article Title: The chemokine receptor CCR10 promotes inflammation-driven hepatocarcinogenesis via PI3K/Akt pathway activation

doi: 10.1038/s41419-018-0267-9

Figure Lengend Snippet: Following short-term DEN-induced inflammation (10 days after i.p. DEN injection), ( a ) Western blotting analysis showed significantly enhanced TNF protein expression, CCR10 protein expression, PI3K protein expression, Akt phosphorylation, and PCNA protein expression. Knocking-out CCR10 significantly opposed these inflammation-induced effects but did not significantly affect TNF or PI3K protein expression.* P < 0.05 vs. vehicle WT group, † P < 0.05 vs. DEN-treated WT group. b Western blotting analysis of CCR10 protein expression, PI3K protein expression, Akt phosphorylation, and PCNA protein expression in murine liver tissue 6 h after intraperitoneal (i.p.) injection of TNF, which produced significant increases in CCR10 protein expression, PI3K protein expression, Akt phosphorylation, and PCNA protein expression. Pretreatment with the CCR10 agonist-ligand CCL28 significantly increased Akt phosphorylation and PCNA expression levels, while pretreatment with the Akt inhibitor A6730 produced the opposite effects. Neither CCL28 nor A6730 had any significant effect upon CCR10 or PI3K expression. * P < 0.05 vs. vehicle group, † P < 0.05 vs. TNF group. All values are reported as means ± standard errors of the mean (SEMs). n = 12 mice in each group

Article Snippet: For some in vitro experiments, HepG2 and LO2 cell lines were pre-treated with the pro-inflammatory cytokine TNF (concentrations as indicated; R&D Systems, Minneapolis, MN, USA) for 4 h, recombinant human CC chemokine ligand 28 (CCL28) (400 nM; R&D Systems) for 2 h, or the allosteric Akt inhibitor A6730 (10 μM; Sigma, St. Louis, MO, USA) for 2 h . Human CCL28 is a natural ligand-agonist for human CCR10 .

Techniques: Injection, Western Blot, Expressing, Phospho-proteomics, Produced