Consistent induction of chronic experimental autoimmune encephalomyelitis in C57BL/6 mice for the longitudinal study of pathology and repair

Abstract

Background While many groups use experimental autoimmune encephalomyelitis (EAE) as a model to uncover therapeutic targets and understand the pathology underlying multiple sclerosis (MS), EAE protocol variability introduces discrepancies in central nervous system (CNS) pathogenesis and clinical disease, limiting the comparability between studies and slowing much-needed translational research. Optimized method Here we describe a detailed, reliable protocol for chronic EAE induction in C57BL/6 mice utilizing two injections of myelin oligodendrocyte glycoprotein (35–55) peptide mixed with Complete Freund’s Adjuvant and paired with pertussis toxin. Results The active MOG35–55 EAE protocol presented here induces ascending paralysis in 80–100% of induced mice. We observe: (1) consistent T cell immune activation, (2) robust CNS infiltration by peripheral immune cells, and (3) perivascular demyelinating lesions concurrent with axon damage in the spinal cord and various brain regions, including the optic nerve, cortex, hippocampus, internal capsules, and the cerebellum. Comparison with existing method(s) Lack of detailed protocols, combined with variability between laboratories, make EAE results difficult to compare and hinder the use of this model for therapeutic development. We provide the most detailed active MOG35–55-EAE protocol to date. With this protocol, we observe high disease incidence and a consistent, reliable disease course. The resulting pathology is MS-like and includes optic neuritis, perivascular mononuclear infiltration, CNS axon demyelination, and axon damage in both infiltrating lesions and otherwise normal-appearing white matter. Conclusions By providing a detailed active MOG35–55-EAE protocol that yields consistent and robust pathology, we aim to foster comparability between pre-clinical studies and facilitate the discovery of MS therapeutics.

Analogues of ERβ ligand chloroindazole exert immunomodulatory and remyelinating effects in a mouse model of multiple sclerosis

“…IndCl has been shown to reduce motor disability in EAE mice when administered prophylactically or therapeutically 10 . Having established that IndCl- o -Cl and IndCl- o -Me exhibited comparable effects to IndCl in vitro , their impact was next evaluated in vivo using eight-week-old female C57BL/6 mice in which EAE had been induced following an established protocol 26 . As a positive control for non-ER isoform specific estrogenic signaling, mice were given prophylactic E2 subcutaneously at the time of initial immunization with MOG 35–55 peptide, which continued throughout the course of experiments (PreEAE + E2 group).…”

Clonal Glial Response in a Multiple Sclerosis Mouse Model

“…In fact, neocortical lesions have previously been recognized to be present in MS (Stadelmann et al, 2008 ). In particular, chronic EAE induced by MOG peptide in mice showed, although with some variability, intracortical MS-like lesions characterized by diffused demyelination, astroglial and microglial activation, oligodendrocyte precursor cells (OPC) proliferation, and a low leukocyte/macrophage infiltrate (Girolamo et al, 2011 ; Mangiardi et al, 2011 ; Hasselmann et al, 2017 ).…”

Increase in chemokine CXCL1 by ERβ ligand treatment is a key mediator in promoting axon myelination

“…Treatment with IndCl and WAY significantly decreased disease severity vs. vehicle beginning by day 24 through 40 (IndCl-purple stars; WAY-orange stars). Differences in EAE clinical scores were determined by two-way unbalanced ANOVA with Dunnett’s multiple comparisons test ( 48 ). n = 8–10 mice per group; * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.…”

“…Active EAE was induced in 8-wk-old female C57BL/6 mice, as previously described ( 14 , 48 ) ( SI Appendix , Fig. S1 ).…”

“…All data were analyzed in GraphPad Prism 6. Differences in EAE clinical scores were determined by two-way unbalanced ANOVA with Dunnett’s multiple comparisons test ( 48 ). All others were analyzed by ordinary one-way ANOVA with Dunnett’s multiple comparisons test or Kruskal–Wallis with Dunn’s multiple comparisons test.…”