30 days after pathogen stimulation, full thickness tail skin grafts were transplanted onto the dorsal thorax of inoculated recipient mice (37 (link)). Skin graft recipients then received either no treatment, CoB consisting of human CTLA4-Ig (500 μg, Bristol-Myers Squibb) and hamster anti-mouse CD154 mAb (500 μg, MR-1, Bio X Cell), rat anti-LFA-1 mAb alone (250 μg, M17/4, Bio X Cell), rat anti-VLA-4 mAb alone (250 μg, PS/2, Bio X Cell), or CoB plus anti-LFA-1 or anti-VLA-4. All mAbs were administered i.p. on post-transplant days 0, 2, 4 and 6. Anti-LFA-1 and anti-VLA-4 mAbs were non-depleting (25 ).
M17 4
Manufactured by BioXCell
The M17/4 is a versatile lab equipment designed for a range of research and analytical applications. It serves as a compact and robust centrifuge, capable of efficiently separating and isolating components from various sample types. The M17/4 features an easy-to-use interface and a reliable performance, making it a useful tool in laboratory settings.
Automatically generated - may contain errors
Lab products found in correlation
Rat igg2b clone ltf 2, by BioXCell (1 mentions)
Anti mouse cd4 mab, by BioXCell (1 mentions)
6 ohda, by Merck Group (1 mentions)
Amd3100, by Merck Group (1 mentions)
Fty720, by Merck Group (1 mentions)
Ovalbumin (ova), by Merck Group (1 mentions)
Poly 1 c, by InvivoGen (1 mentions)
Ova257 264, by GenScript (1 mentions)
Cd8 2.43, by BioXCell (1 mentions)
αcd40 fgk4, by BioXCell (1 mentions)
Brefeldin a, by Thermo Fisher Scientific (1 mentions)
αcd11α, by Merck Group (1 mentions)
Magnetic bead, by Miltenyi Biotec (1 mentions)
7 protocols using m17 4
1
Skin Graft Tolerance by Costimulation Blockade
2
Polymicrobial Sepsis Induction via CLP in Mice
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All the experimental procedures complied with institutional and ARRIVE
guidelines [29 (link)] regarding the use of
animals in research, and were approved by Boston Children’s Hospital
animal care and use committee. Polymicrobial abdominal sepsis was induced by CLP
surgery, as previously described [3 (link),25 (link)]. Briefly, mice were anesthetized with
60 mg/kg ketamine and 5 mg/kg xylazine given intraperitoneally. Following
exteriorization, the cecum was ligated at 1.0 cm from its tip and subjected to a
single, through and through puncture using an 18-gauge needle. A small amount of
fecal material was expelled with gentle pressure to maintain the patency of
puncture sites. The cecum was inserted into the abdominal cavity. 0.1 mL/g of
warmed saline was administered subcutaneously. Buprenorphine was given
subcutaneously to alleviate postoperative surgical pain. Some groups of mice
were placed on a nose cone to be continuously exposed to 1% isoflurane
using isoflurane vaporizer (VetQuip; New South Wales, Australia) for 2 hours.
Isoflurane is often used at the concentration of 1–2% in
clinical practice. Mice were euthanized at indicated time points and were
subjected to analysis. In some experiments, LFA-1 blocking antibody (M17/4;
BioXcell, West Lebanon, NH) 2 mg/kg was given intravenously prior to CLP surgery
as we previously described [30 (link)].
guidelines [29 (link)] regarding the use of
animals in research, and were approved by Boston Children’s Hospital
animal care and use committee. Polymicrobial abdominal sepsis was induced by CLP
surgery, as previously described [3 (link),25 (link)]. Briefly, mice were anesthetized with
60 mg/kg ketamine and 5 mg/kg xylazine given intraperitoneally. Following
exteriorization, the cecum was ligated at 1.0 cm from its tip and subjected to a
single, through and through puncture using an 18-gauge needle. A small amount of
fecal material was expelled with gentle pressure to maintain the patency of
puncture sites. The cecum was inserted into the abdominal cavity. 0.1 mL/g of
warmed saline was administered subcutaneously. Buprenorphine was given
subcutaneously to alleviate postoperative surgical pain. Some groups of mice
were placed on a nose cone to be continuously exposed to 1% isoflurane
using isoflurane vaporizer (VetQuip; New South Wales, Australia) for 2 hours.
Isoflurane is often used at the concentration of 1–2% in
clinical practice. Mice were euthanized at indicated time points and were
subjected to analysis. In some experiments, LFA-1 blocking antibody (M17/4;
BioXcell, West Lebanon, NH) 2 mg/kg was given intravenously prior to CLP surgery
as we previously described [30 (link)].
3
Immune Cell Depletion Prior to Platelet Transfusion
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MZB cells were depleted 48 hours before platelet transfusion by intraperitoneal injection of an anti-mouse CD11a mAb (100 μg, clone M17/4; Bioxcell, Lebanon, NH) and an anti-mouse CD49d mAb (100 μg, clone PS/2; Bioxcell) or isotype-matched control antibodies (rat immunoglobulin G2a (IgG2a), clone 2A3, and rat IgG2b, clone LTF-2; Bioxcell) diluted in phosphate buffered saline.13 (link) Depletion of MZB cells was assessed by quantifying CD45+ B220+ IgM+ CD23− CD21+ splenocytes by flow cytometry.
CD4+ T cells were depleted 4 and 2 days before platelet transfusion by intraperitoneal injection of anti-mouse CD4 mAb (250 μg, clone GK1.5; BioXcell) or isotype-matched control antibodies (rat IgG2b, clone LTF-2; Bioxcell) diluted in phosphate buffered saline. The depletion of CD4+ T cells was assessed by quantifying CD45+ CD4+ cells in the blood and spleen using flow cytometry. The efficiency of CD4+ T-cell depletion was assessed in peripheral blood samples with APC rat anti-mouse CD4 (clone RM-45) (supplemental Data 7 ).
CD4+ T cells were depleted 4 and 2 days before platelet transfusion by intraperitoneal injection of anti-mouse CD4 mAb (250 μg, clone GK1.5; BioXcell) or isotype-matched control antibodies (rat IgG2b, clone LTF-2; Bioxcell) diluted in phosphate buffered saline. The depletion of CD4+ T cells was assessed by quantifying CD45+ CD4+ cells in the blood and spleen using flow cytometry. The efficiency of CD4+ T-cell depletion was assessed in peripheral blood samples with APC rat anti-mouse CD4 (clone RM-45) (
4
Polymicrobial Sepsis Induction via CLP in Mice
5
Lymphocyte Trafficking Regulation via Cell Adhesion and Adrenergic Signaling
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Lymphocyte entry to LNs was blocked by i.v. injection of neutralizing antibodies against integrin αL (M17/4) and α4 (PS/2, both from Bio X Cell) at 100 µg each per mouse and, 14–22 h later, lymphocytes remaining in the LNs were enumerated (Lo et al., 2005 (link)). Mini osmotic pumps (2001D; Alzet) containing saline, AMD3100 (40 mg/kg/d; Sigma-Aldrich) and/or various doses of clenbuterol or salbutamol were surgically implanted s.c. in the back of mice to continuously administer the drugs up to 24 h. Entry blockade was performed 1 h after implantation of the pumps. For depletion of adrenergic nerves, mice were injected i.p. with 100 mg/kg 6-OHDA (Sigma-Aldrich) dissolved in saline containing 0.01% antioxidant ascorbate on days −7 and −5 and 200 mg/kg on day −3 (Grebe et al., 2010 (link)). For short-term transfers, mutant or matched control spleen cells were labeled with 0.5 µM CFSE in DMEM containing 1% FBS for 10 min at 37°C. Each recipient mouse received 2 × 107 WT or Adrb2−/− cells, or 4 × 107Ccr7−/− cells to overcome a defect in LN entry caused by CCR7 deficiency (Förster et al., 1999 (link)). After 2 d of equilibration, entry blockade was performed.
6
Tumor Vaccination and Immune Cell Depletion
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Tumor bearing mice were vaccinated i.p. with 100 µg αCD40 (FGK45; BioXcell), 75 µg polyI:C (Invivogen), and 500 µg ovalbumin (Sigma) or 200 µg OVA257-264 (Genscript) at day ten after tumor injection. FTY720 (Sigma) was provided in the drinking water (2 µg/mL) starting at day nine after tumor implantation and supplemented with daily i.p. injection (25 µg) on days nine through twelve. Depletion antibodies (250 µg) for CD8 (2.43; BioXCell) and/or CD4 (GK1.5; BioXCell) were administered i.p. at day eight after tumor injection and again every four days for the remainder of the experiment. FTY720 and depletion efficiency were confirmed by flow cytometric analysis of blood in each experiment. LFA blocking antibody (M17/4; BioXCell) was administered i.p. (200 µg) every two days. Brefeldin A (Thermo Fisher) was injected i.v. (250 µg) five hours prior to harvest to block cytokine secretion. For OT1 transfer, spleens were harvested from OT1 mice originally from Taconic and maintained at the University of Virginia.
7
Multicolor Tracking of Diabetogenic T Cells
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