Journal: Cell Death & Disease

Article Title: CCR5 signaling promotes lipopolysaccharide-induced macrophage recruitment and alveolar developmental arrest

doi: 10.1038/s41419-021-03464-7

Figure Lengend Snippet: A Representative images of migrating RAW264.7 cells treated with LPS (1 µg/µl) and/or CCL3 (0.1 µg/µl), and DAPTA (1 µg/µl) for 12 h. Top: Scale bar: 100 μm. Bottom: Scale bar: 50 μm. B Representative images of migrating macrophages from BALF after treatment with LPS (1 µg) and/or DAPTA (1 μg) for 72 h. Scale bar: 50 μm. Quantification of transwell migrated macrophages. Data are expressed as mean ± SD ( n = 7), * p < 0.05.

Article Snippet: The saline control group received 5 μL of normal saline per amniotic sac, the LPS group while the LPS, LPS with BTA, LPS with DAPTA, LPS with anti IL-1β, LPS with GSK872 and IL-1β groups received 1 μg of LPS (Escherichia coli 055: B5; Sigma-Aldrich, St. Louis, MO, USA), 1 μg of LPS and 0.4 μg of BTA (Bachem, Bubendorf, Switzerland), 1 μg of LPS and 1 μg of DAPTA (Tocris Bioscience, Bristol, UK), 1 μg of LPS and 0.05 μg of anti-IL-1β (PeproTech, Rocky Hill, NJ, USA), 2 μg of GSK872 (Selleckchem,Houston, TX, USA), and 0.5 μg of IL-1β (PeproTech), respectively, diluted in 5 μL normal saline per amniotic sac.

Techniques:

Journal: Life sciences

Article Title: Involvement of CD4 + and CD8 + T-lymphocytes in the modulation of nociceptive processing evoked by CCL4 in mice.

doi: 10.1016/j.lfs.2022.120302

Figure Lengend Snippet: Fig. 1. A) Thermal hyperalgesia evoked by the administration of 100 ng/kg of CCL4 (s.c.; 1 h before testing) and its prevention by the CCR5 selective antagonist DAPTA (5 mg/kg, s.c., 1 h before testing). (N = 5–6). **P < 0.01 compared with saline (SAL)-treated group, Tukey's test. On top, a diagram of the experimental design is shown. B) Mice received the i.p. administration of anti-CD3 antibody (1 μg/mouse; N = 5) or the corresponding IgG2bκ isotype (1 μg/mouse; N = 5) and 24 h later, withdrawal latencies were taken before any further treatment (BASAL). Next, mice received the acute administration of CCL4 (100 ng/kg) and 1 h later withdrawal latencies were measured again (n = 5). **P < 0.01 compared with the corresponding basal latencies, Bonferroni's correction. On top, a diagram of the experimental design is shown. C) Effect of i.p. treatment 24 h before with either anti-CD3 antibody (1 μg/mouse; N = 5) or IgG2bκ isotype (1 μg/mouse; N = 5) on the total number of circulating white blood cells, lymphocytes, mid-size cells and granulocytes. Individual data are represented as red (anti-CD3 antibody) or blue (isotype) circles and their mean and corresponding S.E. appears as a short red or blue line, respectively. **P < 0.01 compared with isotype-treated group, Bonferroni's correction. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: The CCR5 antagonist DAPTA (5 mg/kg; Tocris), the opioid receptor antagonist naloxone (3 mg/kg; Tocris) and the selective δ-opioid receptor antagonist naltrindole (1 mg/kg; Tocris) were dissolved in saline and administered s.c. 1 h, 15 min and 30 min before testing, respectively.

Techniques: Saline

Journal: Life sciences

Article Title: Involvement of CD4 + and CD8 + T-lymphocytes in the modulation of nociceptive processing evoked by CCL4 in mice.

doi: 10.1016/j.lfs.2022.120302

Figure Lengend Snippet: Fig. 2. Determination by flow cytometry of CCR5 surface expression on CD3+, CD4+ and CD8+ T-cell subsets coming from peripheral blood collected from naïve mice. Histograms of the left-hand side depict representative examples of the fluorescence distribution (mean fluorescence intensity, MFI) detected in the APC channel (CCR5-APC) in cells coming from one sample showing the fluo rescence for the FMO sample (without CCR5 staining) and its CCR5-stained counterpart obtained in CD3+ (A), CD4+ (B) and CD8+ (C) cells. The graphs of the right-hand side show percentages of cells expressing CCR5 on CD3+ (A), CD4+ (B) and CD8+ (C). Dots correspond to individual values ob tained in each sample and red lines represent the calculated mean and the corresponding S.E. (n = 9). (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: The CCR5 antagonist DAPTA (5 mg/kg; Tocris), the opioid receptor antagonist naloxone (3 mg/kg; Tocris) and the selective δ-opioid receptor antagonist naltrindole (1 mg/kg; Tocris) were dissolved in saline and administered s.c. 1 h, 15 min and 30 min before testing, respectively.

Techniques: Flow Cytometry, Expressing, Fluorescence, Staining

Journal: Life sciences

Article Title: Involvement of CD4 + and CD8 + T-lymphocytes in the modulation of nociceptive processing evoked by CCL4 in mice.

doi: 10.1016/j.lfs.2022.120302

Figure Lengend Snippet: Fig. 9. Graphic explanation of the mechanisms proposed to be involved in the switch from analgesia to hyperalgesia when systemic doses of CCL4 are increased from pg/kg to ng/kg. The administration of 100 ng/kg of CCL4 leads to CCR5 activation and the subsequent release of IL-16 from circulating CD8+ T-cells. The activation by IL-16 of CD4 receptors expressed in CD4+ T-cells could evoke CCR5 desensitization in this lymphocyte subset, thus impeding the release of met-enk responsible on the analgesia evoked by lower doses of CCL4 [3]. Besides, the blood increase of hypernociceptive mediators, such as CCL2, CXCL1 and IL-1α [5] provokes a hyperalgesic response.

Article Snippet: The CCR5 antagonist DAPTA (5 mg/kg; Tocris), the opioid receptor antagonist naloxone (3 mg/kg; Tocris) and the selective δ-opioid receptor antagonist naltrindole (1 mg/kg; Tocris) were dissolved in saline and administered s.c. 1 h, 15 min and 30 min before testing, respectively.

Techniques: Activation Assay