Dear Sadia Shaukat

I briefly wanted to reach out concerning your inquiry to give you some pointers for your experiment.

1. The first question is which mouse strain you tried to induce EAE to as there is strict MHCII restriction in mice. For a quick concise introduction on the animal model and the susceptible mouse strains, please see;

Thesis The Genetics of Experimental Arthritis in Rodents

Subsequently, I would have a quick look at the MHCII of your mice and confirm that they indeed harbour susceptibility alleles with basic microsatellite or SNP marker screening.

2. You might want to check the preparation of your CFA peptide emulsion. For some best practices, please see:

Article Standardization of Antigen-Emulsion Preparations for the Ind...

In addition, ensure that you use CFA with Mycobacterium tuberculosis (H37Ra strain), as that influences your outcome in terms of disease severity.

3. Consider alternative mouse strains, e.g. H-2r (MHC) bearing mice and MBP (as these are usually more susceptible). For an example, please see:

Article Polymorphic estrogen receptor binding site causes Cd2-depend...

4. And lastly, please consider you may actually have discovered the effect of a novel (spontaneous) mutation or of an (accidental) crossbreeding in your colony that introduced protective alleles. We had this case years ago in our rat colony and found a mutation that completely abrogated autoimmune rodent models (albeit the initial mutation arose in rats). For the details and especially for the potential sequence of experiments to do, please see:

Article Endophilin A2 deficiency protects rodents from autoimmune ar...

I hope that these considerations help.

Good luck with your experiment,

Michael

It could be due to the quality of the antigen, the adjuvant used, or the method of administration. It could also be due to the age or strain of the mice used in the experiment.

Review your experimental protocol and ensure that you are following it correctly. Orr consult a colleague or mentor who has experience with EAE induction.