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 1

Prevalence of etx in the gut microbiome of people with MS and people who were HCs.

Options: View larger image (or click on image) Download as PowerPoint
Prevalence of etx in the gut microbiome of people with MS and people who...
(A) Workflow of the experimental setup. Fecal bacteria were purified by density gradient centrifugation in a Nycodenz solution. (B) Initial screen by PCR targeting the 3′-terminal sequence of etx (542 bp) detected more frequent presence of etx in people with MS compared with people who were HCs. (C) Confirmatory assays for etx+ participants from the initial screen (B) using PCR for C. perfringens-specific genes. PCRs targeting the 5′-terminal sequence of etx (679 bp) confirmed the initial detection results. Detection of C. perfringens-specific 16S rRNA and the chromosomally located alpha toxin (cpa/plc) gene present in all C. perfringens strains corroborates the source of the etx gene. Participants labeled in red are included as known etx– samples from B and included for comparison. Reference strains including C. perfringens type B (ATCC3626), type D (FD203), and type F (ATCC12915) were included as additional controls. Types B and D are etx-harboring strains and type F is negative for etx. Types B, D, and F are all positive for cpa/plc and C. perfringens-specific 16S rRNA. (D) Statistical analysis of etx frequency by Fisher’s exact test. Similar results were achieved in 3 independently repeated PCR experiments using the same people.