Epsilon toxin–producing Clostridium perfringens colonize the multiple sclerosis gut microbiome overcoming CNS immune privilege
Yinghua Ma, … , Christopher E. Mason, Timothy Vartanian
Yinghua Ma, … , Christopher E. Mason, Timothy Vartanian
Published February 28, 2023
Citation Information: J Clin Invest. 2023;133(9):e163239. https://doi.org/10.1172/JCI163239.
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Epsilon toxin–producing Clostridium perfringens colonize the multiple sclerosis gut microbiome overcoming CNS immune privilege

Abstract

Multiple sclerosis (MS) is a complex disease of the CNS thought to require an environmental trigger. Gut dysbiosis is common in MS, but specific causative species are unknown. To address this knowledge gap, we used sensitive and quantitative PCR detection to show that people with MS were more likely to harbor and show a greater abundance of epsilon toxin–producing (ETX-producing) strains of C. perfringens within their gut microbiomes compared with individuals who are healthy controls (HCs). Isolates derived from patients with MS produced functional ETX and had a genetic architecture typical of highly conjugative plasmids. In the active immunization model of experimental autoimmune encephalomyelitis (EAE), where pertussis toxin (PTX) is used to overcome CNS immune privilege, ETX can substitute for PTX. In contrast to PTX-induced EAE, where inflammatory demyelination is largely restricted to the spinal cord, ETX-induced EAE caused demyelination in the corpus callosum, thalamus, cerebellum, brainstem, and spinal cord, more akin to the neuroanatomical lesion distribution seen in MS. CNS endothelial cell transcriptional profiles revealed ETX-induced genes that are known to play a role in overcoming CNS immune privilege. Together, these findings suggest that ETX-producing C. perfringens strains are biologically plausible pathogens in MS that trigger inflammatory demyelination in the context of circulating myelin autoreactive lymphocytes.

Authors

Yinghua Ma, David Sannino, Jennifer R. Linden, Sylvia Haigh, Baohua Zhao, John B. Grigg, Paul Zumbo, Friederike Dündar, Daniel Butler, Caterina P. Profaci, Kiel Telesford, Paige N. Winokur, Kareem R. Rumah, Susan A. Gauthier, Vincent A. Fischetti, Bruce A. McClane, Francisco A. Uzal, Lily Zexter, Michael Mazzucco, Richard Rudick, David Danko, Evan Balmuth, Nancy Nealon, Jai Perumal, Ulrike Kaunzner, Ilana L. Brito, Zhengming Chen, Jenny Z. Xiang, Doron Betel, Richard Daneman, Gregory F. Sonnenberg, Christopher E. Mason, Timothy Vartanian

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Figure 4

Comparison of ETX- and PTX-EAE in the spinal cord using the classical scoring scale.

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Comparison of ETX- and PTX-EAE in the spinal cord using the classical sc...
(A) Classical EAE time-course (left panel) of mice from indicated experimental groups. Mice immunized with CFA/PBS (PBS) or CFA/MOG35-55 (MOG) were injected i.p. with PTX at 5 μg/kg, ETX at 50 ng/kg (ETX-Lo), or ETX at 500 ng/kg body weight (ETX-Hi) on days 0 and 2 following immunization. Quantification of day of disease onset, peak EAE score, and increasing EAE score (right panels). (B) Representative sections of lumbar spinal cord from mice sacrificed at day 30 stained with Luxol Fast Blue (myelin stains blue). Red circles and black arrowheads indicate demyelination in the dorsal column (dc) or in the lateral and ventral funiculi, respectively. Dashed pink lines indicate lesion border in ventral white matter (WM) tracts. Quantification of myelin integrity (%, middle panel) generated from binary images (right panel). (C) Electron microscopy (left panel) and quantification (middle and right panels) evaluating demyelination in ETX-EAE mice. Red asterisks indicate demyelinated axons; yellow rectangles highlight ultrastructural changes of myelin sheath; cyan arrowhead indicate decompacted myelin sheath; yellow arrowheads indicate splitting myelin sheath and debris; red arrow indicates expanded space between an axon and myelin. Data in left panel of A are mean ± SEM; data in right panels of A and B represent median ± range; Kruscal-Wallis test (nonparametric); NS, not significant; n = 4 mice for PBS > PTX and PBS > ETX-Hi controls, n = 6 mice for MOG > EAE-Lo, and n = 8 mice groups MOG > PTX and MOG > ETX-Hi. Data in C represent mean ± SD; unpaired t test; *** P < 0.001; n = 24 fields from 2 mice. Scale bars: 1 mm (B), 500 nm (C, top panels), and 200 nm (C, lower panels). Similar results for 2 independent experiments.