TY  - JOUR
TI  - Advancing EAE Modeling: Establishment of a Non-Pertussis Immunization Protocol for Multiple Sclerosis
AU  - Gupta, Shruti 
AU  - Arnab, Sreejita 
AU  - Stehle, Elena 
AU  - Nguyen, Kayla L.
VL  - 16
IS  - 3
PY  - 2026
DA  - 2026/02/05
SP  - e5589
C1  - Bio-protocol 2026;16:e5589
DO  - 10.21769/BioProtoc.5589
UR  - https://doi.org/10.21769/BioProtoc.5589
AB  - Experimental autoimmune encephalomyelitis (EAE) is a widely used rodent model of multiple sclerosis (MS), typically induced with pertussis toxin (PTX) to achieve robust disease onset. However, PTX has been shown to exert broad immunomodulatory effects that include disruption of G protein-coupled receptor (GPCR) signaling, altered T-cell response, and exogenous suppression of regulatory T cells, all of which are not present in human MS pathophysiology. Moreover, PTX also obscures the sex differences observed in MS, limiting the translational value of EAE models that rely on it. Given EAE’s widespread use in preclinical therapeutic testing, there is a critical need for a model that better recapitulates both clinical and immunological features of MS without PTX-induced confounds. Here, we demonstrate a non-pertussis toxin (non-PTX) EAE model in C57BL/6 mice, using optimized concentrations of complete Freund’s adjuvant (CFA), Mycobacterium tuberculosis, and myelin oligodendrocyte glycoprotein (MOG35-55) peptide. This model recapitulates hallmark features of MS that include demyelination, neuroinflammation, motor deficits, and neuropathic pain. Importantly, it retains sex-specific differences in disease onset and pathology, providing a more physiologically and clinically relevant platform for mechanistic and translational MS research.
KW  - Pertussis toxin (PTX)
KW  - EAE
KW  - Multiple sclerosis
KW  - Non-PTX EAE
KW  - Neuropathic pain
KW  - Ataxia
JF  - Bio-protocol
SN  - 2331-8325
PB  - Bio-protocol LLC.
BIO101  - False