In Vivo Activation of Peritoneal Macrophages in Response to Antibodies in a Murine Model
Published: August 31, 2023
Abstract
Source: Kozicky, L., et al., Assessment of Antibody-based Drugs Effects on Murine Bone Marrow and Peritoneal Macrophage Activation. J. Vis. Exp. (2017)
This video demonstrates the activation of a mouse's peritoneal macrophages using intravenous immunoglobulin and bacterial lipopolysaccharide. The co-stimulation activates the peritoneal macrophages into anti-inflammatory macrophages possessing immune regulatory activity.
Protocol
All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board. 1. Challenging Mice in Vivo with intravenous immunoglobulin and lipopolysaccharide (IVIg+LPS) and Harvesting Peritoneal Macrophages Weigh each mouse. Calculate the volume of IVIg (100 mg/mL stock concentration) needed to provide a final dose of 2.5 g/kg body weight. Calculate the amount of LPS (100 μ…
Divulgazioni
The authors have nothing to disclose.
Materials
| Iscove's modified Dulbecco's medium (IMDM) | Life technologies | 12440053 | |
| Fetal Bovine Serum (FBS) | Life technologies | 12483-020 | |
| Penicillin-streptomycin | Life technologies | 15140148 | |
| 1X red blood cell lysis buffer | eBioscience | ||
| Cell dissociation buffer | Life technologies | 13150016 | Enzyme-Free, Hanks's-based, EDTA |
| Lipopolysaccharide (LPS) | Sigma aldrich | L 4516 | From E. coli 0127:B8 |
| IVIg (Gammunex) | Grifols | Received from BC Children's Hospital, Transfusion Medicine | |
| IVIg (Gammagard liquid) | Baxter Healthcare Corporation | Received from BC Children's Hospital, Transfusion Medicine | |
| IVIg (Octagam) | Octapharma | Received from BC Children's Hospital, Transfusion Medicine | |
| Phosphate-buffered saline (PBS) (sterile), pH 7.4 | Life technologies | 10010023 | |
| Mouse IL-10 ELISA | BD biosciences | 555252 | |
| Mouse IL-12/23p40 ELISA | BD biosciences | 555165 | |
| 26 g needle | BD biosciences | 305110 | |
| 1 mL syringe | BD biosciences | 309659 | |
| 10 mL syringe | BD biosciences | 309604 | |
| 15 mL conical tube | BD biosciences | 352096 | |
| Forceps | VWR | 82027-386 | Fine tip, dissecting |
| Scissors | VWR | 82027-582 | Delicate, 4 1/2" |
| Brightfield microscope | Motic | AE31 | Inverted phase contrast |
| Scale | Mettler | PE 3000 |
Tags
Citazione di questo articolo
In Vivo Activation of Peritoneal Macrophages in Response to Antibodies in a Murine Model. J. Vis. Exp. (Pending Publication), e21607, doi: (2023).