Preparing an Antigen-Adjuvant Emulsion to Induce Autoimmune Encephalomyelitis
Published: August 31, 2023
Abstract
Source: Bäckström, B. T. A Rapid, Simple, and Standardized Homogenization Method to Prepare Antigen/Adjuvant Emulsions for Inducing Experimental Autoimmune Encephalomyelitis. J. Vis. Exp. (2022)
This video demonstrates an efficient homogenization technique to generate an antigen-adjuvant emulsion to induce experimental autoimmune encephalomyelitis (EAE) in a murine model. The emulsion contains an autoantigen — myelin oligodendrocyte glycoprotein (MOG) — in the water phase, surrounded by a continuous oil phase comprising an inactivated pathogenic bacteria. The emulsion ensures a sustained release of antigens for a prolonged immune response.
Protocol
All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board. NOTE: A schematic flow of the method is described in Figure 1. 1. Material preparation NOTE: Prepare all the reagents aseptically in a sterile hood, and aliquot and store at the indicated temperature. The reagents can be stored for…
Representative Results
Figure 1: Schematic of the process of emulsion preparation using a commercial emulsion kit. This figure has been reused from Topping et al with permission. Figure 2: Characterization of emulsions produced using different methods. (A) Representative images from three different preparation meth…
Disclosures
The authors have nothing to disclose.
Materials
| 1 mL Injection syringe | B. Braun | 9166017V | |
| 1 mL Injection syringe | Sigma-Aldrich | Z683531 | |
| 7 ml empty tubes with caps | Bertin-Instruments | P000944LYSK0A.0 | 7 mL tube |
| 50 mL sterile centrifuge tube | Fisher Scientific | 10788561 | 50 mL tube |
| Dispersant, light mineral oil | Sigma-Aldrich | M8410 | Store at RT |
| Emulsion kit | Bertin-Instruments | D34200.10 ea | Containing a tube, cap, and plunger |
| Incomplete Freund's Adjuvant | Sigma-Aldrich | F5506 | Store at +4 °C |
| Mycobacterium tuberculosis, H37RA | Fisher Scientific | DF3114-33-8 | Store at +4 °C |
| Minilys-Personal homogenizer | Bertin-Instruments | P000673-MLYS0-A | Shaking homogenizer |
| MOG 35-55 Peptide | Innovagen | N/A | |
| Montanide ISA 51 VG | Seppic | 36362Z | FDA-approved oil adjuvant |
| Pall Acrodisc Syringe Filters 0.2 μm | Fisher Scientific | 17124381 | Sterile filter |
| PBS, Ca2+/Mg2+ free | Thermo Fisher Scientific | 14190144 | PBS |
| Phase-Contrast Microscope | Olympus | BX40-B | |
| Steel Beads 3.2 mm | Fisher Scientific | NC0445832 | Autoclave and store at RT |
Tags
Cite This Article
Preparing an Antigen-Adjuvant Emulsion to Induce Autoimmune Encephalomyelitis. J. Vis. Exp. (Pending Publication), e21568, doi: (2023).