Oral Administration of Recombinant Lactococcus lactis Expressing HSP65 and Tandemly Repeated P277 Reduces the Incidence of Type I Diabetes in Non-Obese Diabetic Mice

Abstract

Diabetes mellitus type 1 (DM1) is an autoimmune disease that gradually destroys insulin-producing beta-cells. We have previously reported that mucosal administration of fusion protein of HSP65 with tandem repeats of P277 (HSP65-6P277) can reduce the onset of DM1 in non-obese diabetic (NOD) mice. To deliver large amounts of the fusion protein and to enhance long-term immune tolerance effects, in the present study, we investigated the efficacy of using orally administrated L. lactis expressing HSP65-6P277 to reduce the incidence of DM1 in NOD mice. L. lactis strain NZ9000 was engineered to express HSP65-6P277 either constitutively or by nisin induction. After immunization via gavage with the recombinant L. lactis strains to groups of 4-week old female NOD mice for 36 weeks, we observed that oral administration of recombinant L. Lactis resulted in the prevention of hyperglycemia, improved glucose tolerance and reduced insulitis. Immunologic analysis showed that treatment with recombinant L. lactis induced HSP65- and P277- specific T cell immuno-tolerance, as well as antigen-specific proliferation of splenocytes. The results revealed that the DM1-preventing function was in part caused by a reduction in the pro-inflammatory cytokine IFN-γ and an increase in the anti-inflammatory cytokine IL-10. Orally administered recombinant L. lactis delivering HSP65-6P277 may be an effective therapeutic approach in preventing DM1.

The gut microbiota and Type 1 Diabetes

“…Robert and coworkers demonstrated that a combination of genetically modified Lactococcus lactis that express GAD65 and IL-10 and short-term anti-CD3 treatment reversed diabetes in NOD mice [91]. This and other studies [90, 92] using engineered Lactococcus lactis suggest that genetically modified probiotics are safe and effective vectors for the treatment of T1D.…”

“…Novel therapeutic strategies have been designed to overcome this issue by using engineered Lactococcus lactis bacteria as a safe and effective method to transport autoantigens to the immune system to induce tolerance [90-92]. Robert and coworkers demonstrated that a combination of genetically modified Lactococcus lactis that express GAD65 and IL-10 and short-term anti-CD3 treatment reversed diabetes in NOD mice [91].…”

RECTA: Regulon Identification Based on Comparative Genomics and Transcriptomics Analysis

“…Genes from each CEM were first mapped to the identified operons to retrieve the basic transcription units. Next, 300 bps in the upstream of the translation starting sites for each operon were extracted, in which motif finding was carried out using the web server DMINDA (Ma, Zhang, et al 2014, Yang et al 2017). DMINDA is a dominant motif prediction tool, embraced five analytical algorithms to find, scan, and compare motifs (Li, Liu, Ma and Xu 2011, Liu et al 2017, Ma, Liu, Zhou, Yin, Li and Xu 2013), including a phylogenetic footprint framework to elucidate the mechanism of transcriptional regulation at a system level in prokaryotic genomes (Li, Ma, Mao, Yin, Zhu and Xu 2011, Liu, Zhang, Zhou, Li, Fennell, Wang, Kang, Liu and Ma 2016, Liu, Zhou, Li, Zhang, Zeng, Liu and Ma 2016).…”

“…It is has been widely applied in dairy fermentations, such as cheese and milk product (Wegmann et al 2007). Several studies have provided evidence of its essential roles in wrapping and delivering proteins or vaccinations for immune treatment, such as diabetes (Ma, Liu, et al 2014), malaria (Ramasamy et al 2006), tumors (Bermudez-Humaran et al 2005, Zhang et al 2016) and infections (Hanniffy et al 2007). Holding the advantage of higher acid tolerance to protect vectors from resolving during delivery inside of the animal body, L. lactis has more potential and safety in oral drug development (Hols et al 1999).…”

Recombinant Probiotics and Microbiota Modulation as a Good Therapy for Diseases Related to the GIT

“…Although many studies have focused on the use of recombinant bacteria for the treatment of IBDs, as was previously discussed, the use of recombinant probiotic strains expressing/delivering therapeutic molecules has been explored for treatment or prevention of other diseases such as mucositis, cancer, obesity, multiple sclerosis, and diabetes [182][183][184][185].…”