#2704 Effects of high dietary salt on immunome and microbiome composition – results from a randomized clinical trial

Abstract

Abstract Background and Aims Increased dietary salt intake ranks among the most prominent nutritional risk factors worldwide and has been tied to arterial hypertension and all-cause mortality. The mechanisms responsible are incompletely understood and have recently been extended to include inflammatory and microbiome-associated mechanisms. Building on our preliminary data, the present study investigates whether a moderate increase in salt intake in healthy subjects primes host physiology to a more transiently unstable state, potentially leading to changes in the microbiome-immune axis. Method We conducted a prospective, randomized, double-blinded trial to evaluate the effect of high dietary salt in healthy participants (NCT03024567), where the intervention group (n = 19) was given 6 g of salt (NaCl capsules) in addition to the habitual salt intake, resulting in an average doubling of the salt intake recommended by expert societies. A placebo group (n = 19) received gelatin capsules for 14 days. Clinical parameters, fecal samples and PBMC were collected at baseline and day 14. Shotgun metagenomic sequencing, metabolomics (stool and serum), and single-cell sequencing (Cellular Indexing of Transcriptomes and Epitopes by Sequencing (CITEseq)) of whole PBMCs and CD4+ sorted T cells (∼150 k single cells) was performed. Results Microbiome analysis confirms previous findings showing an increased dissimilarity of microbiome composition (Jensen-Shannon Divergence) under a high-salt. Differential abundance of the microbiome's functional space, as quantified by functional gut-specific modules, picked up a salt-specific module (M00494, NatK-NatR (sodium extrusion) two-component regulatory system) in the salt intervention group. CITEseq revealed a significant reduction of circulating NK cells and an increase in classical monocytes as well as changes to several T cell subsets. Furthermore, within several immune cell subsets, we detected differently expressed genes (DEGs) solely in the salt group. We found 602 DEG in naïve conventional T cells (Tconv) and 202 DEGs in non-naïve Tconv, e.g. belonging to NF-kB and T cell receptor signaling pathways. Conclusion Our placebo-controlled study is the first to utilize different omics techniques to investigate and identify high-salt-induced changes in the microbiome and immunome in healthy individuals that may be relevant for health-to-disease transition.