Dendritic cells treated by Trichinella spiralis muscle larval excretory/secretory products alleviate TNBS-induced colitis in mice

https://doi.org/10.1016/j.intimp.2019.02.028Get rights and content

Highlights

  • T. spiralis MLES (Ts-MLES) regulated DCs phenotype.

  • Ts-MLES-DC ameliorated the severity of the TNBS-induced colitis

  • Ts-MLES-DC inhibited Th1 immune response while enhancing the Th2 and Treg immune response in TNBS-induced colitis in vivo.

Abstract

Background

Therapeutic potential of helminth have been shown to have a protective effect on immune-mediated diseases such as Crohn's disease (CD), which is associated with increased production of T helper cell type 1. However, helminth therapy is unacceptable to patients due to side-effects and the fear of parasites. As helminths regulate the cellular immune responses through innate cells such as dendritic cells (DCs), cellular immunotherapy has been considered a therapeutic option to treat CD.

Methods

Bone marrow-dendritic cells were generated, enriched and treated with Trichinella spiralis muscle larval excretory/secretory products (Ts-MLES). DCs maturation was measured by flow cytometry and cytokine production of DCs were measured by ELISA. Colitis was generated by intrarectal administration of 2,4,6-trinitrobenzene sulfonic acid (TNBS) solution. For adoptive transfer, Ts-MLES treated-DCs injected intravenously 24 h prior to TNBS challenge. Disease activity index (DAI) including weight loss, diarrhea, and bloody stool were measured. Colon segments were stained with hematoxylin and eosin (H.E.) and periodic acid schiff (PAS) staining for histological damage scoring. The relative mRNA expression of cytokines in colon was analyzed by RT-PCR. Cytokine production in colon was measured by ELISA. Splenocytes were separated and cytokine profiles including Th1 (IFN-γ), Th2 (IL-4, IL-13), and Treg subsets (IL-10, TGF-β) were analyzed by flow cytometry.

Results

Ts-MLES regulated the maturation and cytokine production of DCs. Ts-MLES -DC ameliorated the severity of the TNBS-induced colitis. In the colon and the spleen, Ts-MLES-DC decreased IFN-γ (Th1) significantly and increased Th2 (IL-4, IL-13)- and Treg (IL-10, TGF-β)- related cytokines.

Conclusions

Ts-MLES-DC ameliorated the severity of the TNBS-induced colitis through decreasing IFN-γ. Ts-MLES-DC skewed the Th1-mediated response toward the Th2 type and regulatory T cell response.

Section snippets

Background

Inflammatory bowel disease (IBD) is characterized by a chronic relapsing inflammatory condition of the gastrointestinal tract. IBD primarily encompasses ulcerative colitis (UC) and Crohn's disease (CD) [1]. CD is associated with increased production of T helper cell type 1 (Th1)-like cytokines such as Interferon (IFN)-γ. Numerous epidemiological studies have shown that there is an inverse relationship between the prevalence of CD and exposure to helminthic parasites in developing countries and

Animals

BALB/c mice (female, 6–8 weeks old) were purchased from the Shanghai SLAC Company. Female Wistar rats were purchased from the Experimental Animal Centre of College of Basic Medical Sciences, Jilin University (Changchun, China). All animal experiments were performed according to regulations of the Administration of Affairs Concerning Experimental Animals in China. The protocol was approved by the Institutional Animal Care and Use Committee of Jilin University (20170318).

Parasites and preparation of ES

The T. spiralis isolate

T. spiralis MLES (Ts-MLES) regulated DCs phenotype

To investigate the ability of the Ts-MLES to regulate the maturation of DCs, surface costimulatory molecules on CD11c+ DCs (>90% CD11c+, Fig. 1A) were measured. As shown in Fig. 1B and C, the Ts-MLES-treated group showed small increases in CD40, CD80 and CD86 expression compared to the PBS control group. In the absence of Ts-MLES, surface markers (CD40, CD80 and CD86) on CD11c+ DCs stimulated by LPS\IFN-γ were upregulated. However, the Ts-MLES-treated group had significantly inhibited

Discussion

Inflammatory bowel disease (IBD) is a chronic dysregulated inflammatory disease of intestinal tract. The patients frequently experience continuous or intermittent diarrhea, abdominal pain, rectal bleeding and fatigue due to aberrant intestinal inflammation, probably resulting from inappropriately vigorous immune responses to components of the natural intestinal fecal stream [28]. The available treatments for the disease are far from optimal. Several studies have demonstrated the therapeutic

Conclusion

We found that the administration of Ts-MLES-DC alleviated the severity of TNBS-induced colitis in mice. Moreover, our data demonstrated that Ts-MLES-DC modulated inflammatory cytokine production of colon, which resulted in ameliorating intestinal inflammation. The results suggested that Ts-MLES-DC inhibited Th1 immune response while enhancing the Th2 and Treg immune response in TNBS-induced colitis in vivo. These results provide evidence that Ts-MLES-DCs have the potential to mitigate CD via

Conflicts of interest

Authors declare no conflict of interest.

Acknowledgements

We thank Xinrui Wang for the technical assistance. Our thanks are also extended to express our gratitude to all the people who made this work.

Funding

This study was supported by The National Key Research and Development Program of China (2017YFC1601206, 2017YFD0501302, 2017YFD0501300); Jilin Provincial Science and Technology Development Project (20180520042JH); National Natural Science Foundation of China (NSFC 31520103916).

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