We used Maraviroc, a selective CCR5 antagonist, and J-113863, a selective CCR1 antagonist, from Tocris Bioscience (Bristol, UK) and dexamethasone from Sigma Chemical Co (St. Louis, MO, USA). Maraviroc (10 mg/Kg in saline solution), J-113863 (10 mg/Kg in saline solution), or dexamethasone (1 mg/Kg in saline solution) were administered by gavage once a day for 3 days starting at day 9, 48 h after the viral infection, 60 min before the first Cs exposure of the day.
Maraviroc
Manufactured by Bio-Techne
Sourced in United Kingdom, United States
Maraviroc is a laboratory reagent used in scientific research. It is a small-molecule inhibitor that binds to the CCR5 receptor, which is a co-receptor for the human immunodeficiency virus (HIV). Maraviroc can be used in in vitro and in vivo studies to investigate the role of the CCR5 receptor in various biological processes.
Automatically generated - may contain errors
Lab products found in correlation
Bms ccr2 22, by Bio-Techne (2 mentions)
J113863, by Bio-Techne (1 mentions)
Dexamethasone (dex), by Merck Group (1 mentions)
Sialidase, by Merck Group (1 mentions)
Pertussis toxin, by R&D Systems (1 mentions)
Rpmi 1640, by R&D Systems (1 mentions)
Rpmi 1640, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Gemini Bio (1 mentions)
Incb3284, by Bio-Techne (1 mentions)
Phenylephrine, by Merck Group (1 mentions)
Bicuculline, by Merck Group (1 mentions)
Anti p70s6k, by Cell Signaling Technology (1 mentions)
Anti p p70s6k, by Cell Signaling Technology (1 mentions)
Amg487, by Bio-Techne (1 mentions)
Anti lsd1, by Santa Cruz Biotechnology (1 mentions)
Anti akt, by Cell Signaling Technology (1 mentions)
Anti β actin, by Santa Cruz Biotechnology (1 mentions)
Anti erk, by Cell Signaling Technology (1 mentions)
Anti p akt, by Cell Signaling Technology (1 mentions)
Gsk lsd1, by Merck Group (1 mentions)
7 protocols using maraviroc
1
Evaluating CCR5 and CCR1 Antagonists in Viral Infection
2
Sialidase Treatment and Chemokine Receptor Blocking for CD8+ T Cell Analysis
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For removing sialic acid residues, cell-sorted subsets of CD8+ T cells were treated with 0.1 units of sialidase (from Vibrio cholera, Sigma-Aldrich) in 1 ml RPMI 1640 (Life Technologies) containing 10% FBS (Gemini Bio-Products) and 2 mM L-glutamine for 2.5 hr at 37°C. For inhibiting Gi/o proteins, CD8+ T cells were pre-incubated with pertussis toxin (1 μg/ml (R and D Systems) in RPMI 1640 medium containing 10% FBS and 10 mM HEPES for 3 hr at 37°C. For blocking CCR2 and CCR5, pre-incubation was with BMS CCR2 22 (2 μM (Tocris, Minneapolis, MN) or Maraviroc (10 μM (Tocris), respectively, for 30 min at 37°C and inhibitors were left in the medium throughout the assay. For neutralizing CCL20, HUVEC monolayers in flow chambers were pre-treated for 2 hr at 37°C with 20 μg/ml anti-human CCL20/MIP-3α antibody (c67310; R and D Systems), and antibody was maintained at 10 ng/ml throughout the assay.
3
Pharmacological Evaluation of INCB3284 and Maraviroc
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INCB3284 and Maraviroc were purchased from Tocris (Minneapolis, MN). Phenylephrine was obtained from Millipore Sigma (St. Louis, MO).
4
Neurochemical Modulation of Brain Function
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The following drugs were used in the present study: Recombinant CCL3-Macrophage Inflammatory Protein 1 alpha (MIP-1α) (Peprotech Inc. Rocky Hill, NJ, USA), the CCR5 antagonist Maraviroc (Tocris Bioscience, Bristol, UK), the GABAA receptor antagonist, Bicuculline (Sigma Aldrich, Saint Quentin Fallavier, France), the AMPA receptor antagonist 2,3-dioxo-6-nitro-1,2,3,4-tetrahydrobenzoquinoxaline-7-sulfonamide (NBQX) (Tocris Bioscience, Bristol, UK). All agents were diluted directly in the aCSF from stock solutions prepared in distilled water or in dimethylsulfoxide.
5
CHMP4A Knockdown and Overexpression Analysis
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ShCHMP4A was cloned into pSIH‐H1 puro (System Biosciences). The target sequence of CHMP4A short hairpin RNA (shRNA) was 5′‐GCCTAGTGTACCTTCTACT‐3′. FLAG‐tagged CHMP4A eukaryotic expression vector was constructed by inserting PCR‐amplified CHMP4A fragment into pcDNA3.0‐FLAG vector (Invitrogen).
Trizol reagent was purchased from Invitrogen. AMG487 (CXCR3 antagonist) and maraviroc (CCR5 antagonist) were purchased from Tocris. RNase A and GSK‐LSD1 (LSD1 inhibitor) were obtained from Sigma Aldrich. Anti‐dsRNA (J2) was purchased from English and Scientific Consulting Kft. Anti‐CHMP4A was purchased from Sino Biological. Anti‐Flag, Anti‐LSD1, and Anti‐β‐actin were purchased from Santa Cruz Biotechnology. Anti‐p70S6k, Anti‐p‐p70S6K, Anti‐mTOR, Anti‐p‐mTOR, Anti‐Akt, Anti‐p‐Akt, Anti‐ERK, and Anti‐p‐ERK were purchased from Cell Signaling Technology. Neutralizing Anti‐IFNβ, Anti‐PTEN, Anti‐P53, Anti‐P38, and Anti‐p‐P38 were purchased from Proteintech. Human IFNβ ELISA Kit was obtained from CUSABIO.
Whole blood samples, anticoagulated by sodium citrate, were collected from six healthy volunteers (50% male, medium age of 32 years, ranging from 25 to 42 years). The study was approved by the Ethics Committee of the Fourth Medical Center of the Chinese PLA General Hospital. Written informed consent was obtained from all volunteers.
Trizol reagent was purchased from Invitrogen. AMG487 (CXCR3 antagonist) and maraviroc (CCR5 antagonist) were purchased from Tocris. RNase A and GSK‐LSD1 (LSD1 inhibitor) were obtained from Sigma Aldrich. Anti‐dsRNA (J2) was purchased from English and Scientific Consulting Kft. Anti‐CHMP4A was purchased from Sino Biological. Anti‐Flag, Anti‐LSD1, and Anti‐β‐actin were purchased from Santa Cruz Biotechnology. Anti‐p70S6k, Anti‐p‐p70S6K, Anti‐mTOR, Anti‐p‐mTOR, Anti‐Akt, Anti‐p‐Akt, Anti‐ERK, and Anti‐p‐ERK were purchased from Cell Signaling Technology. Neutralizing Anti‐IFNβ, Anti‐PTEN, Anti‐P53, Anti‐P38, and Anti‐p‐P38 were purchased from Proteintech. Human IFNβ ELISA Kit was obtained from CUSABIO.
Whole blood samples, anticoagulated by sodium citrate, were collected from six healthy volunteers (50% male, medium age of 32 years, ranging from 25 to 42 years). The study was approved by the Ethics Committee of the Fourth Medical Center of the Chinese PLA General Hospital. Written informed consent was obtained from all volunteers.
6
Binding Assay for CCR5 Receptor
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7
Depletion of Monocyte Chemotactic Chemokines
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To deplete single or a combination of different monocyte chemotactic chemokines, biotinylated anti-human antibody against CCL2, CCL3, CCL4, and CCL8 were added at a concentration of 5 μg/mL to the different CoM. The biotinylated antibodies were incubated with the CoM for one hour at room temperature before addition of magnetic streptavidin beads (Dynabeads MyOne Streptavidin T1, Life Technologies) using 10 9 beads/mL. These were incubated for one hour with the CoM before removing the beads magnetically.
Successful depletion was confirmed by ELISA using paired capture and detection antiboides for each chemokine (eBioscience) according to manufacturer's instructions and analysed using a Multiskan absorbance plate reader (Thermo Labsystems). For chemokine receptor targeting, 5x10 6 cells/mL PBMC were incubated with inhibitors of CCR2 (RS 504393 or BMS CCR2 22, both from Tocris), CCR5 (Maraviroc, Tocris), or CXCR4 (AMD 3465 hexahydrobromide, Tocris), used at 10 nM, for 30 minutes.
Successful depletion was confirmed by ELISA using paired capture and detection antiboides for each chemokine (eBioscience) according to manufacturer's instructions and analysed using a Multiskan absorbance plate reader (Thermo Labsystems). For chemokine receptor targeting, 5x10 6 cells/mL PBMC were incubated with inhibitors of CCR2 (RS 504393 or BMS CCR2 22, both from Tocris), CCR5 (Maraviroc, Tocris), or CXCR4 (AMD 3465 hexahydrobromide, Tocris), used at 10 nM, for 30 minutes.
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