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Updated: Mar 14, 2026
Microbiota Analysis Using Two-step PCR and Next-generation 16S rRNA Gene Sequencing
Published on: October 15, 2019
Associations between the gut microbiota and host immune markers in pediatric multiple sclerosis and controls
Helen Tremlett1, Douglas W Fadrosh2, Ali A Faruqi2
1Faculty of Medicine (Neurology), University of British Columbia, Room S178, 2211 Wesbrook Mall, Vancouver, BC, V6T 2B5, Canada. helen.tremlett@ubc.ca.
Background:
As little is known of association(s) between gut microbiota profiles and host immunological markers, we explored these in children with and without multiple sclerosis (MS).
Methods:
Children ≤18 years provided stool and blood. MS cases were within 2-years of onset. Fecal 16S rRNA gene profiles were generated on an Illumina Miseq platform. Peripheral blood mononuclear cells were isolated, and Treg (CD4+CD25hiCD127lowFoxP3+) frequency and CD4+ T-cell intracellular cytokine production evaluated by flow cytometry. Associations between microbiota diversity, phylum-level abundances and immune markers were explored using Pearson's correlation and adjusted linear regression.
Results:
Twenty-four children (15 relapsing-remitting, nine controls), averaging 12.6 years were included. Seven were on a disease-modifying drug (DMD) at sample collection. Although immune markers (e.g. Th2, Th17, Tregs) did not differ between cases and controls (p > 0.05), divergent gut microbiota associations occurred; richness correlated positively with Th17 for cases (r = +0.665, p = 0.018), not controls (r = -0.644, p = 0.061). Bacteroidetes inversely associated with Th17 for cases (r = -0.719, p = 0.008), not controls (r = +0.320, p = 0.401). Fusobacteria correlated with Tregs for controls (r = +0.829, p = 0.006), not cases (r = -0.069, p = 0.808).
Conclusions:
Our observations motivate further exploration to understand disruption of the microbiota-immune balance so early in the MS course.
Insights
Gut microbiota profiles in children with multiple sclerosis (MS) showed distinct associations with immune markers like Th17 and Tregs. These findings suggest early gut-immune dysregulation in pediatric MS.
Area of Science:
- Microbiome research
- Immunology
- Pediatric neurology
Background:
- The relationship between gut microbiota composition and host immune system markers in children, particularly those with multiple sclerosis (MS), is not well understood.
- Investigating these associations is crucial for understanding the early pathogenesis of MS.
Purpose of the Study:
- To explore the associations between gut microbiota profiles and immunological markers in children with and without MS.
- To identify potential microbial signatures linked to early-stage MS in pediatric populations.
Main Methods:
- Fecal samples were collected for 16S rRNA gene sequencing to analyze gut microbiota diversity and abundance.
- Peripheral blood mononuclear cells were analyzed for regulatory T-cell (Treg) frequency and CD4+ T-cell cytokine production via flow cytometry.
- Statistical analyses, including Pearson's correlation and adjusted linear regression, were used to examine associations between microbiota and immune markers.
Main Results:
- While overall immune markers (Th2, Th17, Tregs) did not differ significantly between MS cases and controls, specific gut microbiota associations were observed in MS cases.
- Microbiota richness positively correlated with Th17 cells in MS cases, whereas the phylum Bacteroidetes showed an inverse association with Th17 cells.
- Fusobacteria abundance correlated with Tregs in controls but not in MS cases, indicating divergent microbial-host immune interactions.
Conclusions:
- The study highlights distinct gut microbiota associations with immune markers in children with early MS, suggesting a potential disruption of the microbiota-immune balance.
- These findings warrant further investigation into the role of the gut microbiome in the early stages of pediatric MS development and progression.
