mcp 1  (R&D Systems)

Journal: Communications Biology

Article Title: Tick saliva reprograms macrophages into immunosuppressive hubs that regulate T-cell immunity in Rhipicephalus microplus infestation

doi: 10.1038/s42003-026-09981-5

Figure Lengend Snippet: A Representative histograms showing the expression intensity of MHC class II on macrophages stimulated with LPS and IFN-γ in the presence or absence of Rm-saliva. B Summary of the MFI of MHC class II on macrophages. C Summary of the proportions of MHC class II⁺ on macrophages. D Representative flow cytometry plots showing co-expression of MHC class II and CD80. E Summary of the frequencies of MHC class II⁺CD80⁺ macrophages. F Representative flow cytometry plots showing co-expression of MHC class II and CD86. G Summary of the frequencies of MHC class II⁺CD86⁺ macrophages. Concentrations of CCL2 ( H ), CCL3 ( I ), CCL4 ( J ), CCL5 ( K ), CCL8 ( L ), CXCL9 (M) in culture supernatants of macrophages stimulated with LPS and IFN-γ in the presence or absence of Rm-saliva. Chemokine concentrations were generally low under unstimulated conditions, with some values falling below the assay’s detection limit. Data represent paired samples from nine individual cattle ( n = 9). * P < 0.05 (Friedman test followed by the Wilcoxon signed-rank test).

Article Snippet: The concentrations of chemokines in macrophage culture supernatants were determined using the BOVINE CCL2, CCL3, CCL4, CCL5, CCL8, and CXCL9 DO-IT-YOURSELF ELISA Kits (Kingfisher Biotech).

Techniques: Expressing, Flow Cytometry

Journal: Brazilian Dental Journal

Article Title: Chemoattractive potential of stem cells from apical papilla in peripheral blood monocytes: an in vitro study

doi: 10.1590/0103-644020256694

Figure Lengend Snippet: Production of CCL2 quantified by ELISA assay in SCAP conditioned medium (CM-SCAP). Bars show the mean and standard deviation of the experiment (N=3) (A). Viability of monocytes stimulated with decreasing dilutions of SCAP supernatant (CM) compared with control (proliferation medium alone). Absorbance (570nm) data obtained from the MTT assay after 24 h of stimulus exposure (B). Fluorescence data obtained by Alamar Blue assay at experimental time of 24 h. Conditioned medium by SCAP (CM-SCAP) diluted ½ in the presence or not of a CCL2 neutralizer (CM-SCAP + α-CCL2) compared to control (proliferation medium alone) and Recombinant Human CCL2 (CCL2 rH) (C). The result showed the mean and standard deviation of the experiments performed in triplicate. Different letters represent statistical differences between groups. (One-Way ANOVA with Tukey's test, p<0.05) (B and C).

Article Snippet: 300 ng/ml of Recombinant Human CCL2 (279-MC-010 R&D Systems) was used for positive control, while α-MEM 10% was the negative control.

Techniques: Enzyme-linked Immunosorbent Assay, Standard Deviation, Control, MTT Assay, Fluorescence, Alamar Blue Assay, Recombinant

Journal: Neuro-Oncology Advances

Article Title: Adoptive cellular therapy prevents reconstitution of myeloid-derived suppressor cells in the glioma tumor microenvironment

doi: 10.1093/noajnl/vdag054

Figure Lengend Snippet: Integrated analysis of chemokine/cytokine protein after 9-Gy myeloablative radiation, HSCs, and adoptive cellular therapy and in vitro migration assays and single-cell RNA sequencing analysis in KR158B glioma. (A) Cytokine and chemokine profiling of glioma protein lysates in the tumor secretome in adoptive cellular therapy–treated gliomas and untreated control. Statistical comparisons were made using the 1-way ANOVA test. (B) CCL12 protein levels in glioma lysates were quantified by ELISA. Statistical analysis was made using the unpaired Student t test. (C) Transcriptomic analysis of chemokine ligands and receptors using nanoString digital spatial profiling. Statistical analysis was performed using the 2-way ANOVA test. (D) In vitro trans-well migration assay of MDSCs migrating in response to glioma conditioned media from excised glioma tissue combined with neutralizing antibodies against CCL2 and/or CCL12. Statistical analysis was performed using the 1-way ANOVA test. (E) ELISA CCL12 protein quantification using immune cell–specific isolates from excised glioma tissue. Statistical analysis was performed using the 1-way ANOVA test. (F) Immune cell subset deconvolution from single-cell RNA sequencing analysis from excised glioma tissue. (G) mRNA expression of CCL12 overlayed on immune cell subset deconvolution using single-cell RNA sequencing analysis from excised glioma tissue. (H) In vitro trans-well migration assay of MDSCs migration in response to immune cell-specific isolates combined with either IgG or CCL12 neutralizing antibody. Statistical comparisons conducted using the 2-way ANOVA test. * P ≤ .05. ** P ≤ .01. *** P ≤ .001. **** P ≤ .0001; error bars: mean ± standard deviation. Abbreviations: ANOVA, analysis of variance; MDSCs, myeloid-derived suppressor cells.

Article Snippet: Recombinant mouse CCL2 (R&D Systems, cat. 479-JE) and CCL12 (R&D Systems, 428-P5) were diluted in the migration buffer and plated at 150 μL/well in the lower Boyden chamber.

Techniques: In Vitro, Migration, Single Cell, RNA Sequencing, Control, Enzyme-linked Immunosorbent Assay, Expressing, Standard Deviation, Derivative Assay

Journal: Neuro-Oncology Advances

Article Title: Adoptive cellular therapy prevents reconstitution of myeloid-derived suppressor cells in the glioma tumor microenvironment

doi: 10.1093/noajnl/vdag054

Figure Lengend Snippet: Association of Ccl2 and macrophage-associated gene expression with clinical outcomes in glioblastoma patients. (A) Kaplan-Meier analysis of disease-free survival based on Ccl2 expression. (B) Kaplan-Meier analysis of overall survival of Cd14 expression. (C) Kaplan-Meier analysis of disease-free survival stratified by Fcgr3a expression. (D) Deconvolution of single-cell RNA sequencing cell subsets in the glioma microenvironment of GBM patients. (E) Single-cell RNA sequencing deconvolution of Ccl2 expression among cell clusters from GBM patients. (F) Deconvoluted quantification of Ccl2 expression from cell clusters identified from GBM patient samples. Statistical comparisons made using the log-rank test with P values presented on the survival graphs along with hazard ratios and P values for hazard ratios.

Article Snippet: Recombinant mouse CCL2 (R&D Systems, cat. 479-JE) and CCL12 (R&D Systems, 428-P5) were diluted in the migration buffer and plated at 150 μL/well in the lower Boyden chamber.

Techniques: Gene Expression, Expressing, Single Cell, RNA Sequencing

Journal: PLOS One

Article Title: Transport of CCL2 across an induced pluripotent stem cell-derived in vitro model of the human blood-brain barrier is heparan sulfate-dependent

doi: 10.1371/journal.pone.0338780

Figure Lengend Snippet: a. 125 I-CCL2 Pe in the presence of 10 µg/mL CCL2 compared to PBS vehicle. b. 131 I-Alb Pe in the presence of 10 µg/mL CCL2 compared to PBS vehicle. c. 125 I-CCL2 Pe corrected for non-specific leakage as quantified by 131 I-Alb Pe in the presence 10 µg/mL CCL2 compared to PBS vehicle. a-c. One differentiation was performed with n = 7-8 transwells per group. **p < 0.01, ***p < 0.001 (Unpaired two-tailed t-test). Means are displayed with their SD.

Article Snippet: The chloramine-T method was used to radioactively label recombinant carrier-free human CCL2 (R & D Systems, 279-MC-050/CF) with 125 I and bovine serum albumin (BSA, Sigma cat no. 17030-100G) with 131 I, as described previously [ ].

Techniques: Two Tailed Test

Journal: PLOS One

Article Title: Transport of CCL2 across an induced pluripotent stem cell-derived in vitro model of the human blood-brain barrier is heparan sulfate-dependent

doi: 10.1371/journal.pone.0338780

Figure Lengend Snippet: a. 125 I-CCL2 Pe in the presence of 20 U/mL heparin or 10 µM RS504393 (CCR2 antagonist) compared to control (“Ctrl”). b. 131 I-Alb Pe in the presence of 20 U/mL heparin or 10 µM RS504393 compared to an untreated control group. c. 125 I-CCL2 Pe corrected for non-specific leakage as quantified by 131 I-Alb Pe in the presence of 20 U/mL heparin or 10 µM RS504393 compared to an untreated control group. a-c. ***p < 0.001, ****p < 0.0001 (One-way ANOVA with Tukey’s multiple comparisons test). d. Comparison of 125 I-CCL2 and 131 I-Alb Pe in the untreated control group. ****p < 0.0001 (Unpaired two-tailed t-test). a-d. One differentiation was performed with n = 5-6 transwells per group. Means are displayed with their SD.

Article Snippet: The chloramine-T method was used to radioactively label recombinant carrier-free human CCL2 (R & D Systems, 279-MC-050/CF) with 125 I and bovine serum albumin (BSA, Sigma cat no. 17030-100G) with 131 I, as described previously [ ].

Techniques: Control, Comparison, Two Tailed Test

Journal: PLOS One

Article Title: Transport of CCL2 across an induced pluripotent stem cell-derived in vitro model of the human blood-brain barrier is heparan sulfate-dependent

doi: 10.1371/journal.pone.0338780

Figure Lengend Snippet: a. Representative immunocytochemistry of Heparan Sulfate clone F58-10E4 (HS(10E4)) (red) and DAPI (blue) in vehicle or HSase I, II, and III-treated iBECs. b-c. Immunofluorescence analysis of HS(10E4) mean fluorescence intensities (MFI) relative to DAPI area (b) or total HS(10E4) area relative to DAPI area (c). d-e. 125 I-CCL2 (d) or 131 I-Alb in the presence of 0.25 U/ml HSase I, II, and III compared to vehicle. f. 125 I-CCL2 Pe corrected for non-specific leakage as quantified by 131 I-Alb Pe in the presence of 0.25 U/ml HSase I, II, and III compared to vehicle. One differentiation was performed with n = 7-8 transwells per group. *p < 0.05, ***p < 0.001 (Unpaired two-tailed t-test). Means are displayed with their SD. Figure created with BioRender.

Article Snippet: The chloramine-T method was used to radioactively label recombinant carrier-free human CCL2 (R & D Systems, 279-MC-050/CF) with 125 I and bovine serum albumin (BSA, Sigma cat no. 17030-100G) with 131 I, as described previously [ ].

Techniques: Immunocytochemistry, Immunofluorescence, Fluorescence, Two Tailed Test

Journal: PLOS One

Article Title: Transport of CCL2 across an induced pluripotent stem cell-derived in vitro model of the human blood-brain barrier is heparan sulfate-dependent

doi: 10.1371/journal.pone.0338780

Figure Lengend Snippet: a. Molecular structure of GalNAz. b. On day 3 post-subculture, transwells were organized into treatment groups such that TEER means were approximately equal and treated with GalNAz or DMSO (vehicle control).Transwells were re-treated on day 6 post-subculture, and the effects on TEER (c), 125 I-CCL2 Pe (d) and 131 I-Alb Pe (e) were measured on day 9 post-subculture. f. For each transwell, 125 I-CCL2 Pe was corrected for non-specific leakage as quantified by 131 I-Alb Pe. b-f. **p < 0.01, ***p < 0.001 (Unpaired two-tailed t-test). One differentiation was performed with n = 9 transwells treated per group. Means are displayed with their SD. Figure created with BioRender.

Article Snippet: The chloramine-T method was used to radioactively label recombinant carrier-free human CCL2 (R & D Systems, 279-MC-050/CF) with 125 I and bovine serum albumin (BSA, Sigma cat no. 17030-100G) with 131 I, as described previously [ ].

Techniques: Control, Two Tailed Test