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 &#956…

Divulgations

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

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Citer Cet Article
In Vivo Activation of Peritoneal Macrophages in Response to Antibodies in a Murine Model. J. Vis. Exp. (Pending Publication), e21607, doi: (2023).

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