Presenter's Name(s)

Josie KennedyFollow

Primary Faculty Mentor Name

Dimitry Krementsov

Project Collaborators

Theresa Montgomery (Graduate Student Mentor)

Graduate Student Mentors

Theresa Montgomery

Status

Undergraduate

Student College

College of Nursing and Health Sciences

Program/Major

Medical Laboratory Science

Primary Research Category

Biological Sciences

Secondary Research Category

Health Sciences

Presentation Title

Isolation and Characterization of Lactobacillus Species Associated with Disease Susceptibility in a Murine Model of Multiple Sclerosis

Time

11:00 AM

Location

Silver Maple Ballroom - Biological Sciences

Abstract

Multiple sclerosis is the most common non-traumatic neurologic disease of young adults, affecting more than 2 million people worldwide. The etiology is complex and multifaceted, with 30% of disease risk arising from genetics and 70% from environmental factors. The microbiome is the community of commensal, symbiotic and pathogenic microorganisms living in our bodies, and has been shown to be significantly altered in MS patients compared to healthy controls. In order to study the connection between the microbiome and MS, 16s rRNA sequencing was carried out on several different laboratory mouse strains carrying different microbiomes and exhibiting differential susceptibility in a model of MS, experimental autoimmune encephalomyelitis (EAE). The 16s rRNA data showed higher amounts of Lactobacillus species in the EAE-susceptible microbiome, along with meta-data analysis pinpointing Lactobacillus reuteri as a species of interest. Through bacterial isolation techniques, we were able to isolate three different bacterial species from the mouse: L. reuteri, L. murinus, and L. johnsonii, with the former two species showing an inverse relationship with each other. We have successfully transplanted these species into germ free or normal mice through a gavage, and their effects on EAE can be studied. Currently, I am working to characterize the soluble mediators produced by these three species using RAW macrophages. By culturing macrophages with bacterial supernatants and stimulating them with lipopolysaccharide (LPS), I will be able to analyze the levels of inflammatory cytokines induced by each species using an ELISA assay. My hypothesis is that higher levels of inflammatory cytokines will be produced when L. reuteri bacterial supernatant is introduced to the macrophages, which reflects the higher level of MS-like disease state seen in mouse experiments.

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Isolation and Characterization of Lactobacillus Species Associated with Disease Susceptibility in a Murine Model of Multiple Sclerosis

Multiple sclerosis is the most common non-traumatic neurologic disease of young adults, affecting more than 2 million people worldwide. The etiology is complex and multifaceted, with 30% of disease risk arising from genetics and 70% from environmental factors. The microbiome is the community of commensal, symbiotic and pathogenic microorganisms living in our bodies, and has been shown to be significantly altered in MS patients compared to healthy controls. In order to study the connection between the microbiome and MS, 16s rRNA sequencing was carried out on several different laboratory mouse strains carrying different microbiomes and exhibiting differential susceptibility in a model of MS, experimental autoimmune encephalomyelitis (EAE). The 16s rRNA data showed higher amounts of Lactobacillus species in the EAE-susceptible microbiome, along with meta-data analysis pinpointing Lactobacillus reuteri as a species of interest. Through bacterial isolation techniques, we were able to isolate three different bacterial species from the mouse: L. reuteri, L. murinus, and L. johnsonii, with the former two species showing an inverse relationship with each other. We have successfully transplanted these species into germ free or normal mice through a gavage, and their effects on EAE can be studied. Currently, I am working to characterize the soluble mediators produced by these three species using RAW macrophages. By culturing macrophages with bacterial supernatants and stimulating them with lipopolysaccharide (LPS), I will be able to analyze the levels of inflammatory cytokines induced by each species using an ELISA assay. My hypothesis is that higher levels of inflammatory cytokines will be produced when L. reuteri bacterial supernatant is introduced to the macrophages, which reflects the higher level of MS-like disease state seen in mouse experiments.