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.
View: Text | PDF
Research Article Autoimmunity Microbiology Article has an altmetric score of 201

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

×

Figure 6

ETX-EAE mice show increased CD4+ lymphocyte infiltration in the cerebellum and thalamus compared with PTX-EAE.

Options: View larger image (or click on image) Download as PowerPoint
ETX-EAE mice show increased CD4+ lymphocyte infiltration in the cerebell...
Sections from mice sacrificed at day 30 after immunization with CFA/MOG35-55 followed by ETX (ETX-EAE) or PTX (PTX-EAE) injection were immunostained with anti-CD4 antibody (brown) and counter-stained with hematoxylin (purple). (A–C) Representative CD4 staining in spinal cord (A), cerebellum (B) and thalamus (C). In ETX-EAE mice, CD4+ cells were found in perivascular cuffs and surrounding parenchyma indicated by arrowheads and shown in boxed regions of B and C. CD4+ cells from PTX-EAE mice were localized to perivascular space or scattered (arrowhead and arrow). Boxed regions shown at higher magnifications to illustrate morphological details. A high magnification micrograph (bottom panel in B) depicts CD4+ staining (black arrow). OT, optic tracts; ML, medial lemniscus; PC, posterior commissure. (D) ImageJ-generated binary drawings on CD4+ distribution in the white matter (WM) of a PTX-EAE spinal cord (SC, orange arrows) and ETX-EAE cerebellum (Ceb). In Ceb, orange arrowheads indicate CD4+ cells confined to a single layer in the meninges (upper boxed region). Perivascular localization of CD4+ cells shown at a higher magnification in the cerebellum of ETX-EAE mice (lower boxed region). GM, gray matter. Scale bars: 1 mm (top row A, and left column B), 2 mm (top and third rows in left column in C), 200 μm (high magnification of boxed regions A–C), 500 μm (D, left column), 100 μm (D, right column), and 50 μm (B, bottom right panel). (E and F) Quantification of number of CD4+ cells (E) and CD4+ perivascular cuffs (F) in indicated CNS regions. Data represent median ± range; Kruscal-Wallis test (nonparametric). NS, not significant. n = 4 and 8 mice for controls (PBS > PTX; PBS > ETX-Hi) and EAE groups (MOG > PTX; MOG > ETX-Hi), respectively. Similar results were obtained from 2 independent experiments.