Oral administration of recombinant Bacillus subtilis spores expressing Helicobacter pylori neutrophil-activating protein suppresses peanut allergy via up-regulation of Tregs

Clin Exp Allergy. 2019 Dec;49(12):1605-1614. doi: 10.1111/cea.13489. Epub 2019 Sep 12.

Abstract

Background: Helicobacter pylori neutrophil-activating protein (NAP) is an immune modulator with anti-Th2 inflammation activity that can be used to prevent IgE-mediated allergic reactions. Cholera toxin B (CTB) is a mucosal adjuvant that can induce antigen tolerance. Bacillus subtilis spores are an ideal vehicle for the oral delivery of heterologous antigens.

Objective: We investigated the therapeutic effect of recombinant NAP B subtilis spores on peanut allergies in a mouse model.

Methods: Female C3H/HeJ mice were sensitized and challenged with peanut extract by oral administration. Before challenge, recombinant NAP and CTB-NAP (CNAP) spores were orally administered to sensitized mice for 4 weeks. Faecal peanut-specific IgA and serum-specific IgE, IgG1, and IgG2a levels were measured, and the intestinal microbiota was analysed. Mice were intraperitoneally injected with anti-CD25 antibodies for regulatory T cell (Treg) depletion to evaluate the efficacy of Tregs in preventing peanut allergy. After challenge, anaphylactic reactions, plasma histamine, Tregs, and splenocyte interleukin (IL)-10, IL-4, IL-5 and interferon-γ (IFN-γ) levels were evaluated.

Results: After 4 weeks of recombinant spore treatment, faecal IgA levels and serum IgG2a levels were increased, while serum IgG1 and IgE levels were reduced. Intestinal microbiota analysis revealed that CNAP spores increased the taxonomic abundance of Firmicutes at the phylum level and Clostridia at the class level. After challenge, the administration of NAP or CNAP spores to mice was found to ameliorate anaphylactic reactions and decrease plasma histamine levels. Administration of NAP or CNAP spores also enhanced IL-10 and IFN-γ secretion, and suppressed IL-4 and IL-5 secretion. The protective effect of CNAP spores was more pronounced than that of NAP spores; this therapeutic effect was lost after Treg depletion.

Conclusions and clinical relevance: Recombinant NAP spores successfully suppressed Th2 inflammation via the up-regulation of Tregs; this may serve as a novel therapeutic approach for treating food allergies.

Keywords: Bacillus subtilis spores; Treg; intestinal microbiota; neutrophil-activating protein; peanut allergy.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Administration, Oral
  • Animals
  • Bacillus subtilis* / genetics
  • Bacillus subtilis* / immunology
  • Bacterial Proteins* / genetics
  • Bacterial Proteins* / immunology
  • Female
  • Helicobacter pylori / genetics*
  • Mice
  • Microorganisms, Genetically-Modified* / genetics
  • Microorganisms, Genetically-Modified* / immunology
  • Peanut Hypersensitivity* / immunology
  • Peanut Hypersensitivity* / therapy
  • Spores, Bacterial* / genetics
  • Spores, Bacterial* / immunology
  • T-Lymphocytes, Regulatory / immunology*
  • T-Lymphocytes, Regulatory / pathology

Substances

  • Bacterial Proteins