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Diabetologia

pp 1–14 | Cite as

Oral butyrate does not affect innate immunity and islet autoimmunity in individuals with longstanding type 1 diabetes: a randomised controlled trial

  • Pieter F. de GrootEmail author
  • Tatjana Nikolic
  • Sultan Imangaliyev
  • Siroon Bekkering
  • Gaby Duinkerken
  • Fleur M. Keij
  • Hilde Herrema
  • Maaike Winkelmeijer
  • Jeffrey Kroon
  • Evgeni Levin
  • Barbara Hutten
  • Elles M. Kemper
  • Suat Simsek
  • Johannes H. M. Levels
  • Flora A. van Hoorn
  • Renuka Bindraban
  • Alicia Berkvens
  • Geesje M. Dallinga-Thie
  • Mark Davids
  • Frits Holleman
  • Joost B. L. Hoekstra
  • Erik S. G. Stroes
  • Mihai Netea
  • Daniël H. van Raalte
  • Bart O. Roep
  • Max Nieuwdorp
Article

Abstract

Aims/hypothesis

The pathophysiology of type 1 diabetes has been linked to altered gut microbiota and more specifically to a shortage of intestinal production of the short-chain fatty acid (SCFA) butyrate, which may play key roles in maintaining intestinal epithelial integrity and in human and gut microbial metabolism. Butyrate supplementation can protect against autoimmune diabetes in mouse models. We thus set out to study the effect of oral butyrate vs placebo on glucose regulation and immune variables in human participants with longstanding type 1 diabetes.

Methods

We administered a daily oral dose of 4 g sodium butyrate or placebo for 1 month to 30 individuals with longstanding type 1 diabetes, without comorbidity or medication use, in a randomised (1:1), controlled, double-blind crossover trial, with a washout period of 1 month in between. Participants were randomly allocated to the ‘oral sodium butyrate capsules first’ or ‘oral placebo capsules first’ study arm in blocks of five. The clinical investigator received blinded medication from the clinical trial pharmacy. All participants, people doing measurements or examinations, or people assessing the outcomes were blinded to group assignment. The primary outcome was a change in the innate immune phenotype (monocyte subsets and in vitro cytokine production). Secondary outcomes were changes in blood markers of islet autoimmunity (cell counts, lymphocyte stimulation indices and CD8 quantum dot assays), glucose and lipid metabolism, beta cell function (by mixed-meal test), gut microbiota and faecal SCFA. The data was collected at the Amsterdam University Medical Centers.

Results

All 30 participants were analysed. Faecal butyrate and propionate levels were significantly affected by oral butyrate supplementation and butyrate treatment was safe. However, this modulation of intestinal SCFAs did not result in any significant changes in adaptive or innate immunity, or in any of the other outcome variables. In our discussion, we elaborate on this important discrepancy with previous animal work.

Conclusions/interpretation

Oral butyrate supplementation does not significantly affect innate or adaptive immunity in humans with longstanding type 1 diabetes.

Trial registration

Netherlands Trial Register: NL4832 (www.trialregister.nl).

Data availability

Raw sequencing data are available in the European Nucleotide Archive repository (https://www.ebi.ac.uk/ena/browse) under study PRJEB30292.

Funding

The study was funded by a Le Ducq consortium grant, a CVON grant, a personal ZONMW-VIDI grant and a Dutch Heart Foundation grant.

Keywords

Butyrate Diabetes Microbiota Short-chain fatty acids 

Abbreviations

CCR

C-C chemokine receptor

CRP

C-reactive protein

CXCR

CXC chemokine receptor

DRiP

Defective ribosomal product

IA

Islet antigen

IMDM

Iscove’s modified Dulbecco’s medium

IQR

Interquartile range

LPS

Lipopolysaccharide

MFI

Mean fluorescence intensity

NK

Natural killer

PBMC

Peripheral blood mononuclear cell

PPI

Preproinsulin

SCFA

Short-chain fatty acid

Treg

T regulatory cell

Notes

Acknowledgements

We cordially thank C. Rustemeijer (Amstelland Hospital, Amstelveen, the Netherlands), V. Gerdes (Amsterdam UMC, Amsterdam, the Netherlands), T. Brouwer (OLVG Hospital, Amsterdam, the Netherlands), S. van Dam (OLVG Hospital, Amsterdam, the Netherlands) and J. Hensbergen (Amsterdam UMC, Amsterdam, the Netherlands) for inclusion of participants.

Contribution statement

PFdeG, FH, JBLH, BOR and MNi designed the study. SS substantially contributed to the acquisition of data. PFdeG, BOR, MNe, ESGS, FMK, DHvR, TN, SB, GD, HH, MW, BH, JK, EMK, JHML, SI, EL, GMD-T, MD, FAvH, RB and AB contributed to the analysis and/or interpretation of data. PFdeG, BOR and MNi drafted the manuscript. All authors critically revised the manuscript. All authors gave their approval of the final (published) version of the manuscript. All authors agree to be accountable for all aspects of the work in ensuring that questions related to the accuracy or integrity of any part of the work are appropriately investigated and resolved. PFdeG is the guarantor of this work and, as such, had full access to all the data in the study and takes responsibility for the integrity of the data and the accuracy of the data analysis.

Funding

This study was supported by Le Ducq consortium grant 17CVD01 (to MNi) and CVON grant 2O18.27 (to SB, MNe and MNi). MNi is supported by a personal ZONMW-VIDI grant 2013 (016.146.327) and a Dutch Heart Foundation grant.

Duality of interest

The authors declare that there is no duality of interest associated with this manuscript.

Supplementary material

125_2019_5073_MOESM1_ESM.pdf (1.8 mb)
ESM (PDF 1793 kb)
125_2019_5073_MOESM2_ESM.csv (1010 kb)
ESM Table 2 (CSV 1009 kb)
125_2019_5073_MOESM3_ESM.csv (1.9 mb)
ESM Table 3 (CSV 1990 kb)

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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2020

Authors and Affiliations

  • Pieter F. de Groot
    • 1
    Email author
  • Tatjana Nikolic
    • 2
  • Sultan Imangaliyev
    • 1
  • Siroon Bekkering
    • 1
    • 3
  • Gaby Duinkerken
    • 2
  • Fleur M. Keij
    • 2
  • Hilde Herrema
    • 1
  • Maaike Winkelmeijer
    • 1
  • Jeffrey Kroon
    • 1
  • Evgeni Levin
    • 1
  • Barbara Hutten
    • 4
  • Elles M. Kemper
    • 5
  • Suat Simsek
    • 6
  • Johannes H. M. Levels
    • 1
  • Flora A. van Hoorn
    • 1
  • Renuka Bindraban
    • 1
  • Alicia Berkvens
    • 1
  • Geesje M. Dallinga-Thie
    • 1
  • Mark Davids
    • 1
  • Frits Holleman
    • 1
  • Joost B. L. Hoekstra
    • 1
  • Erik S. G. Stroes
    • 1
  • Mihai Netea
    • 3
    • 7
  • Daniël H. van Raalte
    • 1
    • 8
  • Bart O. Roep
    • 2
    • 9
  • Max Nieuwdorp
    • 1
    • 8
  1. 1.Department of Internal and Vascular MedicineAcademic Medical CenterAmsterdamthe Netherlands
  2. 2.Department of Immunohematology and Blood TransfusionLeiden University Medical CenterLeidenthe Netherlands
  3. 3.Department of Internal MedicineRadboud University Medical CenterNijmegenthe Netherlands
  4. 4.Department of Epidemiology, Amsterdam University Medical Centers, Academic Medical CentreAmsterdamthe Netherlands
  5. 5.Clinical Pharmacy, Amsterdam University Medical Centers, Academic Medical CentreAmsterdamthe Netherlands
  6. 6.Department of Internal MedicineAlkmaar Medical Center (MCA)Alkmaarthe Netherlands
  7. 7.Department for Genomics & Immunoregulation, Life and Medical Sciences Institute (LIMES)University of BonnBonnGermany
  8. 8.Diabetes Center, Department of Internal Medicine, Amsterdam University Medical Centers, VU University Medical CentreAmsterdamthe Netherlands
  9. 9.Department of Diabetes Immunology, Diabetes & Metabolism Research Institute at the Beckman Research Institute, City of HopeDuarteUSA

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