Epsilon toxin–producing Clostridium perfringens colonize the multiple sclerosis gut microbiome overcoming CNS immune privilege
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
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
View: Text | PDF
Research Article Autoimmunity Microbiology

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 2

etx abundance and etx-harboring strains increase in the fecal microbiota of patients with MS.

Options: View larger image (or click on image) Download as PowerPoint
etx abundance and etx-harboring strains increase in the fecal microbiot...
(A and B) Representative amplification plots (A) and statistical analysis of etx abundance (B) by TaqMan real-time PCR analysis. Of note, the system detected 15 etx+ patients with MS, fewer than what was detected by standard PCR (19). (C–E) cpa abundance in people who were healthy controls (HCs) and patients with MS. (F) Multiplex PCR reveals different composition of strains in the fecal C. perfringens community from participants who were etx+. Participants labeled in red are included as known etx– samples from (1B) and included for comparison. Of note, C. perfringens type E, defined by the presence of cpa and itx, is present in MS3, MS31, and HC1; while MS19, MS26, HC1, and HC4 carry cpb, a virulence determinant carried by C. perfringens types C and B. (G–I) Analysis of the etx/cpa ratio by TaqMan real-time PCR using the C. perfringens type B (ATCC3626) and type D (FD203) strains as a calibrator for the quantification of etx+/etx– strain ratios in the fecal microbiota from etx+ participants. Quantification of the ratio of etx/cpa by 2–ΔCt (G), etx/cpa against type D culture as calibrator by 2–ΔΔCt (H). Dashed horizontal lines in G and H show 2–ΔCt/2–ΔΔCt values from culture of indicated reference strains. Estimate of maximum of percentages of etx– and etx+ strains in etx+ HC and MS fecal microbiome (I). The estimates assume that the individuals with a 2–ΔΔCt value above 1 (type D) contain 100% etx+ strains. P value determined by Mann-Whitney test (non-Gaussian distribution). Number of participants (n) is indicated in individual panels; black lines in B–E, and H indicate medians. Data in G are mean ± SD, n = 3 technical triplicates in a representative real-time PCR experiment). Similar results were achieved in 3 independently repeated PCR/qPCR experiments using the same participants.