Oral administration of recombinant Lactococcus lactis expressing HSP65 and tandemly repeated P277 reduces the incidence of type I diabetes in non-obese diabetic mice

• Published in: PloS one 2014-08, Vol.9 (8), p.e105701-e105701
• Main Authors: Ma, Yanjun, Liu, Jingjing, Hou, Jing, Dong, Yuankai, Lu, Yong, Jin, Liang, Cao, Rongyue, Li, Taiming, Wu, Jie
• Format: Article
• Language: English
• Subjects: Administration, Oral; Animals; Antigens; Atherosclerosis; Beta cells; Binding sites; Biology and Life Sciences; Chaperonin 60 - genetics; Chaperonin 60 - therapeutic use; Cytokines; Cytokines - immunology; Deoxyribonucleic acid; Diabetes; Diabetes mellitus; Diabetes mellitus (insulin dependent); Diabetes Mellitus, Type 1 - epidemiology; Diabetes Mellitus, Type 1 - immunology; Diabetes Mellitus, Type 1 - prevention & control; DNA; E coli; Female; Fusion protein; Glucose; Glucose tolerance; Health aspects; Heat shock proteins; Heat-Shock Proteins - genetics; Heat-Shock Proteins - therapeutic use; Hsp65 protein; Humans; Hyperglycemia; Immunization; Immunological tolerance; Immunotherapy; Incidence; Inflammation; Insulin; Insulitis; Interleukin 10; Laboratories; Lactococcus lactis; Lactococcus lactis - genetics; Life sciences; Lymphocytes T; Medicine and Health Sciences; Mice; Mice, Inbred NOD; Mucosa; Nisin; Obesity; Oral administration; Peptide Fragments - genetics; Peptide Fragments - therapeutic use; Peptides; Pharmaceuticals; Pharmacy; Plasmids; Recombinant Fusion Proteins - genetics; Recombinant Fusion Proteins - therapeutic use; Rodents; Splenocytes; T cells; T-Lymphocytes - cytology; T-Lymphocytes - immunology; Type 1 diabetes; Vaccines; γ-Interferon
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0105701

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.