SB-PEPTIDE offers a turnkey solution for multiple sclerosis (MS) studies with an experimental autoimmune encephalomyelitis kit (EAE kit). Each kit provided allows the induction of EAE in 9 mice (0.2mL/mouse). The kit contains 2.1mL of a ready-to-use MOG35-55/CFA emulsion,one vial containing 5µg pertussis toxin (PTX) in glycerol buffer and a recommended experimental protocol with typical results.
MOG35-55/CFA emulsion
Myelin Oligodendrocyte Glucoprotein (MOG35-55) is used in emulsion with Complete Freund’s Adjuvant (CFA) in EAE studies. Emulsification is an important step in the success of the EAE induction,results will partially depend on this phase. SB-PEPTIDE offers a ready-to-use MOG35-55/CFA emulsion.
The complete Freund’s adjuvant is used as an immunopotentiator,it is composed of inactivated and dried mycobacteria and used with MOG 35-55 to induce EAE. Experimental autoimmune encephalomyelitis is the model commonly used to study efficacy of drugs for treatment of multiple sclerosis (MS) but also pathogenesis of autoimmunity,CNS inflammation,demyelination,cell trafficking and tolerance induction.
The kit contains 3 tubes with 0.7mL of pre-filled MOG/CFA emulsion,one vial containing 5µg pertussis toxin (PTX) in glycerol buffer and a recommended experimental protocol
Recommended syringe and needles (not included): 1mL syringe with needle 19Gx1 1/2″ – 1 ,1 x 40 mm for emulsion suction and 22G x 1 1/4″,0.7 x 30 mm needle for administrations
SB-PEPTIDE offers to prepare customized EAE induction kit,with various antigen peptides (see SB-PEPTIDE’s EAE peptide range here) with specific concentrations of antigen and CFA. Contact us for more details.
References & citations
J Neurosci Methods. 2022 Feb 1;367:109443. doi: 10.1016/j.jneumeth.2021.109443. Epub 2021 Dec 15
Myelin oligodendrocyte glycoprotein induced experimental autoimmune encephalomyelitis (EAE) is the most commonly used animal model of multiple sclerosis. However,variations in the induction protocol can affect EAE progression,and may reduce the comparability of data.
Optimized method
In the present study,we investigated the influence of the different components used for EAE induction in C57BL/6J mice on disease progression. MOG35–55-induced chronic EAE in C57BL/6J mice has been applied as a model to challenge optimal pertussis toxin (PTx) dosing,while considering variations in batch potency.
Results
We demonstrate that the dosage of PTx,adjusted to its potency,influences EAE development in a dose-dependent manner. Our data show that with our protocol,which considers PTx potency,C57BL/6J mice consistently develop symptoms of EAE. The mice show a typical chronic course with symptom onset after 10.5 ± 1.08 days and maximum severity around day 16 postimmunization followed by a mild remission of symptoms.
Comparison with existing methods
Previously studies reveal that alterations in PTx dosing directly modify EAE progression. Our present study highlights that PTx batches differ in potency,resulting in inconsistent EAE induction. We also provide a clear protocol that allows a reduction in the number of mice used in EAE experiments,while maintaining consistent results.
Conclusion
Higher standards for comparability and reproducibility are needed to ensure and maximize the generation of reliable EAE data. Specifically,consideration of PTx potency. With our method of establishing consistent EAE pathogenesis,improved animal welfare standards and a reduction of mice used in experimentation can be achieved.
Application note: Effect of CFA contration on EAE induction
Experimental autoimmune encephalomyelitis (EAE) is one of the most popular animal models of multiple sclerosis (MS). Mouse models of demyelinating diseases have been useful in both the demonstration of T cellmediated demyelination and in the characterization of the pathogenesis of immune-mediated demyelinating disease. EAE is a CD4 + T cell-mediated,demyelinating autoimmune disease of the CNS that is characterized by mononuclear cell infiltration. EAE can be induced by various antigens including MOG35-55,a peptide derived from the Myelin Oligodendrocyte Glycoprotein,a structural protein in the myelin sheath. The injection of this peptide emulsified with CFA in mice induces a production of anti-MOG antibodies that cause demyelination and a chronic EAE. The purpose of this study was to investigate the influence of CFA amount on EAE induction.
BACKGROUND: Myelin oligodendrocyte glycoprotein (MOG) is located on the external surface of myelin,a membranous component of the central nervous system (CNS) that forms the insulating lipid layer around neurons. The major MOG splicing variant (a1 transcript) encodes a transmembrane protein with an extracellular domain of an Ig variable (IgV) fold. MOG IgV domains from the same or different cells dimerize and contribute to the organization and maintenance of the myelin sheath in neurons. The encepalitogenic T cells recognize MOG and its immunodominant epitopes (epitopes 1-22,35-55 and 92-106 located at the dimer interface) as foreign antigens and cause the destruction of myelin (demyelination) leading to the clinical condition known as multiple sclerosis (MS). Recognition of the antigen takes place in the context of the trimolecular complex formed by HLA,MOGpeptides and TCR.
OBJECTIVE: Understanding the role of MOG in MS.
METHOD/RESULTS: We have reviewed herein,the genomic organization of the human MOG gene,the structural characteristics of the MOG protein,the involvement of MOG in MS and clinical studies for the treatment of MS based on MOG peptide analogues.
CONCLUSION: Conjugates of antigenic MOG peptides to mannan and combinations of antigenic MOG and other peptides chemically linked to cells of the immune system may modify the immune response,alleviating in some cases the symptoms of MS.
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