Journal: Nature communications

Article Title: The HIV co-receptor CCR5 regulates osteoclast function.

doi: 10.1038/s41467-017-02368-5

Figure Lengend Snippet: Fig. 7 The blockade of CCL5 in vivo. a The serum levels of CCL3 and CCL5 in 10-week-old mice were measured by an ELISA (n = 5). b Mouse anti-CCL5 neutralizing antibodies (mCCL5 neu ab, 500 μg per mouse) were injected (once per week for 2 weeks) into 6-week-old male C57BL/6J mice (n = 5). IgG was administered to the control group. c, d μCT images (scale bars, 100 μm) and parameters (n = 5 mice per group) are shown. e Representative images of the distal femurs from the control IgG and mCCL5 ab groups. HE- (scale bars, 100 μm) and TRAP-stained sections (scale bars, 50 μm) are shown in the upper and lower panels, respectively (n = 5). f Quantitative bone histomorphometric analyses were conducted of the trabecular bones in the distal femurs of control IgG and mouse CCL5 neutralizing antibody-injected mice. *P < 0.05 (by Student’s t-test) in comparison to control and mCCL5-neuAb mice. All values are shown as the mean ± SD, n = 5

Article Snippet: Recombinant mouse CCL5 (RANTES, 478-MR) and CCL9 (MIP-1γ, 3217-MG), anti-human (MAB182), and anti-mouse (MAB-478) CCL5 neutralizing antibodies were purchased from R&D Systems.

Techniques: In Vivo, Enzyme-linked Immunosorbent Assay, Injection, Control, Staining, Comparison, Neutralizing Assay

Journal: Nature communications

Article Title: Asthma reduces glioma formation by T cell decorin-mediated inhibition of microglia.

doi: 10.1038/s41467-021-27455-6

Figure Lengend Snippet: Fig. 3 Asthma reduces T cell induction of Ccl5 in microglia. a Schematic representation of the mouse Nf1OPG neuron-immune-cancer cell axis in the setting of asthma. b No difference in optic nerve Mdk RNA expression was observed between OVA- (n = 3) or HDM- (n = 3) treated mice and controls (n = 6). Bar graphs represent the means ± SEM of independent biological samples. One-way ANOVA with Bonferroni post hoc correction. c Ccl4 levels are similarly induced with increasing midkine concentrations (0–100 ng/ml) in CD3+ T cells from OVA- and PBS-treated mice. Data are presented as the means ± SEM of n = 3 independent biological samples. One-way ANOVA with Bonferroni post hoc correction. Data are presented as the means ± SEM. d, e Similarly increased CD8+, relative to CD4+, T cell content was detected in the optic nerves of Nf1OPG mice treated with PBS (n = 8), OVA (n = 8) or HDM (n = 8). One-way ANOVA with Bonferroni post hoc correction. Activated T cell medium from (f) OVA- (n = 4) and g HDM- (n = 4) treated mice induced lower levels of microglial Ccl5 relative to PBS-treated Nf1OPG mice. Two-tailed Student’s t test. Similar reductions of Ccl5 expression were observed in CD4+ and CD8+ T cells from (h) OVA- (n = 3) and (i) HDM- (n = 3) treated mice relative to PBS-treated controls (n = 3). One-way ANOVA with Bonferroni post hoc correction. Data are presented as the means ± SEM. d Scale bars, 40 µm. From left to right in each panel: b ns; c ns; ns; ns; ns; e P < 0.0001, P = 0.0022, P = 0.0011; f P = 0.0008; g P = 0.0015; h P = 0.0186, P = 0.0054; i P = 0.0224, P = 0.0007.

Article Snippet: In total, 5 × 105 microglia were grown in T cell-conditioned media (TCM) for 48 h8,9, followed Ccl5 (R&D Systems, MMR00) and decorin (Abcam, ab155454) determinations by ELISA.

Techniques: RNA Expression, Two Tailed Test, Expressing

Journal:

Article Title: Release of Macrophage Migration Inhibitory Factor and CXCL8/Interleukin-8 from Lung Epithelial Cells Rendered Necrotic by Influenza A Virus Infection

doi: 10.1128/JVI.76.18.9298-9306.2002

Figure Lengend Snippet: Human primers

Article Snippet: Briefly, 96-well microtiter plates (Maxisorp; Nunc, Wiesbaden, Germany) were coated in phosphate-buffered saline (PBS) with monoclonal antibodies specific for MIF, CXCL8/IL-8, CXCL1/GRO-α, or CCL5/RANTES (R&D Systems, Wiesbaden, Germany) or CCL2/MCP-1 (PharMingen, Hamburg, Germany).

Techniques: Sequencing

Journal:

Article Title: Release of Macrophage Migration Inhibitory Factor and CXCL8/Interleukin-8 from Lung Epithelial Cells Rendered Necrotic by Influenza A Virus Infection

doi: 10.1128/JVI.76.18.9298-9306.2002

Figure Lengend Snippet: Chemokine release from influenza A virus-infected cells a

Article Snippet: Briefly, 96-well microtiter plates (Maxisorp; Nunc, Wiesbaden, Germany) were coated in phosphate-buffered saline (PBS) with monoclonal antibodies specific for MIF, CXCL8/IL-8, CXCL1/GRO-α, or CCL5/RANTES (R&D Systems, Wiesbaden, Germany) or CCL2/MCP-1 (PharMingen, Hamburg, Germany).

Techniques: Virus, Control

Journal:

Article Title: Regulated on Activation, Normal T-Cell-Expressed and -Secreted mRNA Expression in Normal Endometrium and Endometriotic Implants

doi:

Figure Lengend Snippet: Isolated primary endometriotic stromal and epithelial cells were examined for RANTES mRNA expression by in situ hybridization. Subconfluent monolayers of endometriotic stromal cells were cultured for 48 hours. No RANTES mRNA was detectable under conditions in which the medium contained no added cytokine (A). After culturing the cells in the presence of TNF-α (100 ng/ml) for 48 hours (B), RANTES mRNA was expressed in 10 to 20% of the stromal cells. No specific autoradiographic signal was observed when TNF-α stimulated endometriotic stromal cells were hybridized with the RANTES sense RNA-control probe (C). Subconfluent monolayers of epithelial cells did not reveal evidence of RANTES expression, even after TNF-α stimulation (D). Original magnifications, ×600.

Article Snippet: Immunoperoxidase staining was performed overnight at 4°C, using mouse monoclonal IgG antibodies against human cytokeratin-18 (1:2000 dilution; Sigma, St. Louis, MO), human CD-68 (dilution 1:100; DAKO Corp., Carpinteria, CA), human RANTES (dilution 1:100; R&D Systems, Minneapolis, MN), or TNF-α (dilution 1:100, R&D Systems).

Techniques: Isolation, Expressing, In Situ Hybridization, Cell Culture, Control

Journal:

Article Title: Regulated on Activation, Normal T-Cell-Expressed and -Secreted mRNA Expression in Normal Endometrium and Endometriotic Implants

doi:

Figure Lengend Snippet: In situ hybridization of RANTES mRNA in normal mid-proliferative human endometrium (A). RANTES gene expression could be detected in the stromal compartment adjacent to epithelial glands, but the glands themselves did not contain RANTES message. No specific autoradiographic signal was observed when serial tissue sections were hybridized with the RANTES sense RNA-control probe (B). Endometriotic implants also showed RANTES message in the stromal compartment with several intense spots (black arrows) and other more diffuse hybridization (white arrows). The epithelial cell lining of the endometrioma and the unaffected ovarian stroma (top) did not contain RANTES message (C). No specific autoradiographic signal was observed when endometriotic tissue sections were hybridized with the RANTES sense RNA-control probe (D). Immunohistochemical localization of CD68-positive macrophages (E, black arrows) in an adjacent endometriotic tissue section. Monoclonal antibodies against human RANTES demonstrated this protein in the endometriotic stroma, with sparing of the cyst epithelium (F). Monoclonal antibodies against TNF-α showed the protein in the epithelial and stromal compartments of the endometriotic implant. Portions of the ovary uninvolved with endometriosis showed less TNF-α immunostaining (G). Control experiments were performed on serial endometriotic sections stained with TNF-α antibodies immunoabsorbed with excess TNF-α protein (H). Epithelial cells lining the endometriotic cyst were cytokeratin-positive (I). All sections were also counterstained with hematoxylin. H&E staining shows the morphology of the endometriotic implant (K). Original magnifications, ×200.

Article Snippet: Immunoperoxidase staining was performed overnight at 4°C, using mouse monoclonal IgG antibodies against human cytokeratin-18 (1:2000 dilution; Sigma, St. Louis, MO), human CD-68 (dilution 1:100; DAKO Corp., Carpinteria, CA), human RANTES (dilution 1:100; R&D Systems, Minneapolis, MN), or TNF-α (dilution 1:100, R&D Systems).

Techniques: In Situ Hybridization, Gene Expression, Control, Hybridization, Immunohistochemical staining, Bioprocessing, Immunostaining, Staining

Journal: Toxicology and applied pharmacology

Article Title: MUTZ-3 derived Langerhans cells in human skin equivalents show differential migration and phenotypic plasticity after allergen or irritant exposure.

doi: 10.1016/j.taap.2015.05.017

Figure Lengend Snippet: Fig. 3. MUTZ-LC migration out of the epidermis after irritant but not allergen exposure is CCL5 dependent. SE-LC were exposed to water vehicle (V), 10 mM NiSO4 (N) or 2.5 mM Triton X-100 (T) for 16 h. Chemical exposure was performed in the presence of the neutralizing antibody to CCL5 (+) or IgG1 isotype control (−). Number of CD1a-PE positive MUTZ-LC in epidermal sheets was quantified using Image J software. Data represent the average of 4 individual experiments ± SEM. *p b 0.05, **p b 0.01 was calculated using the Mann–Whitney t-test.

Article Snippet: For blocking experiments 7 μg/mL goat anti-human CXCL12 (AF-310-NA, R&D systems, Minneopolis, MN, USA), 7 μg/mL goat anti-human CCL5 (AF-278-NA, R&D systems) or 2 μg/mL goat anti-human IL-10 (AF-217NA, R&D systems) was added to the culture medium prior to chemical exposure (optimal blocking concentration was previously determined) (Ouwehand et al., 2008, 2010a, 2011a).

Techniques: Migration, Control, Software, MANN-WHITNEY