Contributions
Abstract: P446
Type: Poster Sessions
Abstract Category: Pathology and pathogenesis of MS - Microbiology and Virology
Introduction: The microbiome and metagenome are essential to human health and homeostasis. While alterations in the microbiome have been described in MS, there is no demonstrated link between the gut microbiome and MS lesion formation. Clostridium perfringens epsilon toxin is an attractive candidate for new lesion formation as it selectively targets both CNS endothelial cells and oligodendrocytes/myelin.
Objectives: To determine whether MS patients are more likely than healthy controls to be colonized by epsilon toxin-producing strains of C. perfringens.
Aims: 1. Screen for epsilon toxin production in fecal samples from people with MS and healthy controls. 2. Determine the C. perfringens toxinotype in fecal cultures by PCR-based genotyping. 3. Isolation of a pure epsilon toxin-producing strain from MS subjects. 4. Verify that epsilon toxin-producing C. perfringens strains isolated from MS subjects are not laboratory strain contaminants. 5. Preliminary analysis of the genetic composition of the MS-related epsilon toxin plasmid.
Methods: We developed a highly sensitive ELISA for detection of epsilon toxin from clinical samples. Fecal samples from consented subjects were placed in Rapid Perfringens Media for anaerobic culture in a microtitre plate. Supernatants from the microtiter plate were probed by ELISA. Wells corresponding to positive hits by ELISA were then used for PCR based genotyping and multiple rounds of subculture.
Results: 21% of tested MS subjects are colonized by C. perfringens type B in their gut compared to 0% of healthy controls. Sequence analysis confirmed that patient derived type B strains were not due to contamination by the laboratory strain.
Conclusions: Colonization of the gut by C. perfringens type B is statistically associated with MS. While two epsilon toxin-producing C. perfringens strains (type B and type D) exist in nature, thus far we have identified only type B strains from MS subjects.
Disclosure: Research was generously supported by the National Multiple Sclerosis Society and the National Institutes of Health. Funding was provided by U.S. patent No. 9,758,573, ´Methods to Protect Against and Treat Multiple Sclerosis.´
Abstract: P446
Type: Poster Sessions
Abstract Category: Pathology and pathogenesis of MS - Microbiology and Virology
Introduction: The microbiome and metagenome are essential to human health and homeostasis. While alterations in the microbiome have been described in MS, there is no demonstrated link between the gut microbiome and MS lesion formation. Clostridium perfringens epsilon toxin is an attractive candidate for new lesion formation as it selectively targets both CNS endothelial cells and oligodendrocytes/myelin.
Objectives: To determine whether MS patients are more likely than healthy controls to be colonized by epsilon toxin-producing strains of C. perfringens.
Aims: 1. Screen for epsilon toxin production in fecal samples from people with MS and healthy controls. 2. Determine the C. perfringens toxinotype in fecal cultures by PCR-based genotyping. 3. Isolation of a pure epsilon toxin-producing strain from MS subjects. 4. Verify that epsilon toxin-producing C. perfringens strains isolated from MS subjects are not laboratory strain contaminants. 5. Preliminary analysis of the genetic composition of the MS-related epsilon toxin plasmid.
Methods: We developed a highly sensitive ELISA for detection of epsilon toxin from clinical samples. Fecal samples from consented subjects were placed in Rapid Perfringens Media for anaerobic culture in a microtitre plate. Supernatants from the microtiter plate were probed by ELISA. Wells corresponding to positive hits by ELISA were then used for PCR based genotyping and multiple rounds of subculture.
Results: 21% of tested MS subjects are colonized by C. perfringens type B in their gut compared to 0% of healthy controls. Sequence analysis confirmed that patient derived type B strains were not due to contamination by the laboratory strain.
Conclusions: Colonization of the gut by C. perfringens type B is statistically associated with MS. While two epsilon toxin-producing C. perfringens strains (type B and type D) exist in nature, thus far we have identified only type B strains from MS subjects.
Disclosure: Research was generously supported by the National Multiple Sclerosis Society and the National Institutes of Health. Funding was provided by U.S. patent No. 9,758,573, ´Methods to Protect Against and Treat Multiple Sclerosis.´