Calcinaro F, Dionisi S, Marinaro M, Candeloro P, Bonato V, Marzotti S et al.. Oral probiotic administration induces interleukin-10 production and prevents spontaneous autoimmune diabetes in the non-obese diabetic mouse. Diabetologia 48: 1565-1575

Department of Internal Medicine, University of Perugia, Italy.
Diabetologia (Impact Factor: 6.67). 09/2005; 48(8):1565-75. DOI: 10.1007/s00125-005-1831-2
Source: PubMed


Recent observations suggest the involvement of the gastrointestinal tract in the pathogenesis of islet autoimmunity. Thus, the modulation of gut-associated lymphoid tissue may represent a means to affect the natural history of the disease. Oral administration of probiotic bacteria can modulate local and systemic immune responses; consequently, we investigated the effects of oral administration of the probiotic compound VSL#3 on the occurrence of diabetes in non-obese diabetic (NOD) mice.
VSL#3 was administered to female NOD mice three times a week starting from 4 weeks of age. A control group received PBS. Whole blood glucose was measured twice a week. IFN-gamma and IL-10 production/expression was evaluated by ELISA in culture supernatants of mononuclear cells isolated from Peyer's patches and the spleen, and by real-time PCR in the pancreas. Insulitis was characterised by immunohistochemistry and histomorphometric studies.
Early oral administration of VSL#3 prevented diabetes development in NOD mice. Protected mice showed reduced insulitis and a decreased rate of beta cell destruction. Prevention was associated with an increased production of IL-10 from Peyer's patches and the spleen and with increased IL-10 expression in the pancreas, where IL-10-positive islet-infiltrating mononuclear cells were detected. The protective effect of VSL#3 was transferable to irradiated mice receiving diabetogenic cells and splenocytes from VSL#3-treated mice.
Orally administered VSL#3 prevents autoimmune diabetes and induces immunomodulation by a reduction in insulitis severity. Our results provide a sound rationale for future clinical trials of the primary prevention of type 1 diabetes by oral VSL#3 administration.

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    • "It has been hypothesized that beneficial microbes or probiotics can protect against diabetes ( Neu et al . , 2010 ) . VSL#3 has been shown to prevent diabetes in NOD mice when administered after weaning ( Calcinaro et al . , 2005 ) . Yet , admin - istration to germ - free NOD neonatal mice shows no effect ( Yurkovetskiy et al . , 2013 ) . Although NOD mice were deficient in several beneficial microbes present in the probiotic VSL#3 , we found that the administration of VSL#3 failed to protect against accelerated diabetes in NOD mice . Despite VSL#3 being adminis"
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    ABSTRACT: Accumulating evidence supports that the intestinal microbiome is involved in Type 1 diabetes (T1D) pathogenesis through the gut-pancreas nexus. Our aim was to determine whether the intestinal microbiota in the non-obese diabetic (NOD) mouse model played a role in T1D through the gut. To examine the effect of the intestinal microbiota on T1D onset, we manipulated gut microbes by: (1) the fecal transplantation between non-obese diabetic (NOD) and resistant (NOR) mice and (2) the oral antibiotic and probiotic treatment of NOD mice. We monitored diabetes onset, quantified CD4+T cells in the Peyer's patches, profiled the microbiome and measured fecal short-chain fatty acids (SCFA). The gut microbiota from NOD mice harbored more pathobionts and fewer beneficial microbes in comparison with NOR mice. Fecal transplantation of NOD microbes induced insulitis in NOR hosts suggesting that the NOD microbiome is diabetogenic. Moreover, antibiotic exposure accelerated diabetes onset in NOD mice accompanied by increased T-helper type 1 (Th1) and reduced Th17 cells in the intestinal lymphoid tissues. The diabetogenic microbiome was characterized by a metagenome altered in several metabolic gene clusters. Furthermore, diabetes susceptibility correlated with reduced fecal SCFAs. In an attempt to correct the diabetogenic microbiome, we administered VLS#3 probiotics to NOD mice but found that VSL#3 colonized the intestine poorly and did not delay diabetes. We conclude that NOD mice harbor gut microbes that induce diabetes and that their diabetogenic microbiome can be amplified early in life through antibiotic exposure. Protective microbes like VSL#3 are insufficient to overcome the effects of a diabetogenic microbiome.The ISME Journal advance online publication, 14 August 2015; doi:10.1038/ismej.2015.114.
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    • "Recent studies have confirmed that probiotic bacteria could induce regulatory T cells or Th2 cells that could produce anti-inffiammatory cytokines, IL-10 and IL-4 and reduce auto-reactive T-cells, which produce pro-inffiammatory cytokines as interferon γ (IFN-γ) (Matsuzaki et al., 1997; Calcinaro et al., 2005; von Boehmer, 2005; Kwon et al., 2010). Bacillus amyloliquefaciens (BA) is a major workhorse for the production of a variety of extracellular enzymes including phytase, α-amylases, cellulase, metalloproteases and proteases that could enhance digestibility and absorption of nutrients (Gould et al., 1975; Gracia et al., LPS, the primary component of the outer membrane of gram-negative bacteria, serves as a potent activator of innate immune responses that results in the production of pro-and anti-inflammatory mediators (Ulevitch and Tobias, 1995). "
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    • "Multiple low dose streptozotocin induced lymphocyte infiltration in islets of all groups. The degree of infiltration was determined by blinded analysis of insulitis grading [34]. Control and hydrocortisone treated mice showed no signs of lymphocyte infiltration (Figure 6B). "
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    ABSTRACT: Role of GLP-1 and GIP in beta cell compensatory responses to beta cell attack and insulin resistance were examined in C57BL/6 mice lacking functional receptors for GLP-1 and GIP. Mice were treated with multiple low dose streptozotocin or hydrocortisone. Islet parameters were assessed by immunohistochemistry and hormone measurements were determined by specific enzyme linked immunoassays. Wild-type streptozotocin controls exhibited severe diabetes, irregularly shaped islets with lymphocytic infiltration, decreased Ki67/TUNEL ratio with decreased beta cell and increased alpha cell areas. GLP-1 and GIP were co-expressed with glucagon and numbers of alpha cells mainly expressing GLP-1 were increased. In contrast, hydrocortisone treatment and induction of insulin resistance increased islet numbers and area, with enhanced beta cell replication, elevated mass of beta and alpha cells, together with co-expression of GLP-1 and GIP with glucagon in islets. The metabolic responses to streptozotocin in GLP-1RKO and GIPRKO mice were broadly similar to C57BL/6 controls, although decreases in islet numbers and size were more severe. In contrast, both groups of mice lacking functional incretin receptors displayed substantially impaired islet adaptations to insulin resistance induced by hydrocortisone, including marked curtailment of expansion of islet area, beta cell mass and islet number. Our observations cannot be explained by simple changes in circulating incretin concentrations, suggesting that intra-islet GLP-1 and GIP make a significant contribution to islet adaptation, particularly expansion of beta cell mass and compensatory islet compensation to hydrocortisone and insulin resistance.
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