Preparing an Antigen-Adjuvant Emulsion to Induce Autoimmune Encephalomyelitis
Trascrizione
Experimental autoimmune encephalomyelitis, EAE, is a central nervous system disease. Autoantibodies target the myelin oligodendrocyte glycoprotein, MOG, on the myelin sheath of neurons, causing myelin destruction and nerve degeneration.
EAE induction in mice requires injecting an antigen-adjuvant emulsion of the autoantigen MOG mixed with an adjuvant — containing a bacterial antigen — to enhance the immune response.
To prepare the emulsion, take an aqueous suspension of MOG in a homogenizer tube. Place the tube on ice, preventing antigen degradation.
Add the adjuvant — inactivated pathogenic bacteria dispersed in an oil phase. Place the tube inside a shaking homogenizer. The shaking causes turbulence in the fluid, generating an emulsion.
Under a microscope, the emulsion shows uniformly dispersed, micro-sized, aqueous droplets — containing MOG — within a continuous oil phase. The homogeneity of the emulsion ensures higher reproducibility of disease induction.
To inject the emulsion in a susceptible strain of mice, attach a syringe at the front of the tube and transfer the emulsion into it. Take an anesthetized mouse and inject the emulsion subcutaneously.
Upon injection, the oil in the emulsion ensures slow and sustained antigen release, increasing the immune response duration.
The inactivated pathogen in the emulsion causes a localized inflammatory response, recruiting immune cells to the injection site. The recruited immune cells recognize the released MOG antigen and stimulate antibody production, leading to the induction of EAE.
