# HLA-B27-associated gut microbiota and amino acid perturbations promote ankylosing spondylitis through M1 macrophage activation

## TL;DR

This study shows how HLA-B27 and gut bacteria changes lead to ankylosing spondylitis by causing inflammation through M1 macrophages.

## Contribution

The study identifies microbial and metabolic signatures linked to AS and validates a causal role of gut microbiota in disease progression.

## Key findings

- HLA-B27 positivity is associated with altered gut microbiota and amino acid metabolism in AS patients.
- Fecal microbiota transplantation from AS patients induces M1 macrophage activation and disease features in mice.
- Cinnabarinic acid suppresses M1 macrophage polarization via the AhR pathway in AS.

## Abstract

Ankylosing spondylitis (AS) is strongly associated with the human leukocyte antigen B27 (HLA-B27), yet how this genetic risk factor interacts with the gut microbiome remains unclear. We integrated fecal gut microbiota analysis, untargeted metabolomics, and clinical phenotyping in 88 participants, including HLA-B27–positive patients with AS (n = 28), HLA-B27–positive healthy controls (n = 30), and HLA-B27–negative healthy controls (n = 30). HLA-B27 positivity, particularly in AS, was associated with marked alterations in gut microbial composition and metabolic profiles, with forty bacterial species showing progressive disease-related shifts across cohorts. Integrated pathway and metabolomic analyses identified three amino acid–related pathways consistently disrupted in AS: tryptophan metabolism, cysteine metabolism, and pyruvate-centered biosynthesis of branched-chain amino acids, ornithine, and lysine. Correlation network analyses linking differential taxa, metabolites, and clinical indices revealed previously unrecognized microbial and metabolic signatures that robustly distinguished AS from both control groups. To explore causality, fecal microbiota transplantation (FMT) from clinical donors into antibiotic-treated mice recapitulated key disease-relevant features, including impaired intestinal barrier function, systemic inflammation, trabecular bone loss, and polarization of macrophages toward a proinflammatory M1 phenotype. Mechanistic validation identified cinnabarinic acid as a critical microbial-derived metabolite that suppresses M1 macrophage polarization via activation of the aryl hydrocarbon receptor (AhR) pathway and confers protection in the FMT model. Together, these findings support a model in which HLA-B27–associated gut dysbiosis and metabolic reprogramming promote AS pathogenesis through macrophage-mediated inflammation and osteocatabolic signaling, highlighting microbial–metabolic pathways as potential therapeutic targets.