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Diabetologia

, Volume 61, Issue 6, pp 1384–1396 | Cite as

Targeted delivery of antigen to intestinal dendritic cells induces oral tolerance and prevents autoimmune diabetes in NOD mice

  • Yulin Chen
  • Jie Wu
  • Jiajia Wang
  • Wenjing Zhang
  • Bohui Xu
  • Xiaojun Xu
  • Li Zong
Article

Abstract

Aims/hypothesis

The intestinal immune system is an ideal target to induce immune tolerance physiologically. However, the efficiency of oral protein antigen delivery is limited by degradation of the antigen in the gastrointestinal tract and poor uptake by antigen-presenting cells. Gut dendritic cells (DCs) are professional antigen-presenting cells that are prone to inducing antigen-specific immune tolerance. In this study, we delivered the antigen heat shock protein 65-6×P277 (H6P) directly to the gut DCs of NOD mice through oral vaccination with H6P-loaded targeting nanoparticles (NPs), and investigated the ability of this antigen to induce immune tolerance to prevent autoimmune diabetes in NOD mice.

Methods

A targeting NP delivery system was developed to encapsulate H6P, and the ability of this system to protect and facilitate H6P delivery to gut DCs was assessed. NOD mice were immunised with H6P-loaded targeting NPs orally once a week for 7 weeks and the onset of diabetes was assessed by monitoring blood glucose levels.

Results

H6P-loaded targeting NPs protected the encapsulated H6P from degradation in the gastrointestinal tract environment and significantly increased the uptake of H6P by DCs in the gut Peyer’s patches (4.1 times higher uptake compared with the control H6P solution group). Oral vaccination with H6P-loaded targeting NPs induced antigen-specific T cell tolerance and prevented diabetes in 100% of NOD mice. Immune deviation (T helper [Th]1 to Th2) and CD4+CD25+FOXP3+ regulatory T cells were found to participate in the induction of immune tolerance.

Conclusions/interpretation

In this study, we successfully induced antigen-specific T cell tolerance and prevented the onset of diabetes in NOD mice. To our knowledge, this is the first attempt at delivering antigen to gut DCs using targeting NPs to induce T cell tolerance.

Keywords

Autoimmune diabetes Dendritic cells Nanoparticles NOD mice Oral tolerance Oral vaccination 

Abbreviations

CFSE

Carboxyfluorescein succinimidyl ester

CLSM

Confocal laser scanning microscopy

ConA

Concanavalin A

CS

Chitosan

DC

Dendritic cell

FITC-H6P

FITC-labelled heat shock protein 65-6×P277

FRET

Fluorescence resonance energy transfer

H6P

Heat shock protein 65-6×P277

H6P/RMCS

Heat shock protein 65-6×P277-loaded RGD- and mannose-modified chitosan

HSP

Heat shock protein

i.g.

Intra-gastric

MCS

Mannose-modified chitosan

NP

Nanoparticle

RCS

RGD-modified chitosan

RGD

Arginylglycylaspartic acid

RMCS

RGD- and mannose-modified chitosan

SGF

Simulated gastric fluid

SIF

Simulated intestinal fluid

Th

T helper

Treg

Regulatory T cell

Notes

Acknowledgements

We thank Y. Xing (manager of flow cytometry at the China Pharmaceutical University) for her scientific advice and technical assistance with flow cytometry.

Contribution statement

YLC, JW, XJX and LZ contributed to the conception and design of the study. YLC, JJW, WJZ and BHX performed the experiments and analysed the results. YLC, JW, XJX and LZ drafted the manuscript. All authors revised the manuscript critically and gave final approval of the submitted version. LZ is the guarantor of the work.

Funding

This study was supported by National Natural Science Foundation of China (No. 30973650/H3008) and the Postgraduate Research & Practice Innovation Program of Jiangsu Province (KYCX17_0676).

Duality of interest

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

Supplementary material

125_2018_4593_MOESM1_ESM.pdf (268 kb)
ESM (PDF 267 kb)

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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of PharmaceuticsChina Pharmaceutical UniversityNanjingPeople’s Republic of China
  2. 2.Minigene Pharmacy LaboratoryChina Pharmaceutical UniversityNanjingPeople’s Republic of China
  3. 3.State Key Laboratory of Natural MedicinesChina Pharmaceutical UniversityNanjingPeople’s Republic of China

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