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Antibiotics in early life alter the gut microbiome and increase disease incidence in a spontaneous mouse model of autoimmune insulin-dependent diabetes.

Candon S, Perez-Arroyo A, Marquet C, Valette F, Foray AP, Pelletier B, Milani C, Ventura M, Bach JF, Chatenoud L - PLoS ONE (2015)

Bottom Line: In NOD females a slight yet not significant trend towards an increase in disease incidence was observed.Administration of the antibiotic mixture resulted in near complete ablation of the gut microbiota.These results show that a directed even partial ablation of the gut microbiota, as induced by vancomycin, significantly increases type 1 diabetes incidence in male NOD mice thus prompting for caution in the use of antibiotics in pregnant women and newborns.

View Article: PubMed Central - PubMed

Affiliation: Université Paris Descartes, Sorbonne Paris Cité, F-75475, Paris, France; INSERM U1151, Hôpital Necker-Enfants Malades, Paris, France; CNRS UMR 8253, Hôpital Necker-Enfants Malades, Paris, France.

ABSTRACT
Insulin-dependent or type 1 diabetes is a prototypic autoimmune disease whose incidence steadily increased over the past decades in industrialized countries. Recent evidence suggests the importance of the gut microbiota to explain this trend. Here, non-obese diabetic (NOD) mice that spontaneously develop autoimmune type 1 diabetes were treated with different antibiotics to explore the influence of a targeted intestinal dysbiosis in the progression of the disease. A mixture of wide spectrum antibiotics (i.e. streptomycin, colistin and ampicillin) or vancomycin alone were administered orally from the moment of conception, treating breeding pairs, and during the postnatal and adult life until the end of follow-up at 40 weeks. Diabetes incidence significantly and similarly increased in male mice following treatment with these two antibiotic regimens. In NOD females a slight yet not significant trend towards an increase in disease incidence was observed. Changes in gut microbiota composition were assessed by sequencing the V3 region of bacterial 16S rRNA genes. Administration of the antibiotic mixture resulted in near complete ablation of the gut microbiota. Vancomycin treatment led to increased Escherichia, Lactobacillus and Sutterella genera and decreased members of the Clostridiales order and Lachnospiraceae, Prevotellaceae and Rikenellaceae families, as compared to control mice. Massive elimination of IL-17-producing cells, both CD4+TCRαβ+ and TCRγδ+ T cells was observed in the lamina propria of the ileum and the colon of vancomycin-treated mice. These results show that a directed even partial ablation of the gut microbiota, as induced by vancomycin, significantly increases type 1 diabetes incidence in male NOD mice thus prompting for caution in the use of antibiotics in pregnant women and newborns.

No MeSH data available.


Related in: MedlinePlus

Analysis of gene expression in the gut.Relative expression of Foxp3, Ifng, Il10, Il17a and Tgfb1 in the colon, ileum, Peyer’s patches and mesenteric lymph nodes of 10-week-old male (a) and female (b) mice treated with vancomycin (grey bars) and controls (open bars). Five animals per group were analyzed. Expression of Foxp3, Ifng, Il17a and Il10 was normalized to that of Cd3, while expression of Tgfb1 was normalized to Hprt expression. Results are expressed as fold of the mean expression observed in untreated mice. Only changes in expression levels outside the grey zone indicates changes in expression >2-fold increase or decrease in vancomycin-treated mice (black bars) as compared to controls (open bars). Significant differences are indicated as follows: * p<0.05; ** p<0.01.
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pone.0125448.g006: Analysis of gene expression in the gut.Relative expression of Foxp3, Ifng, Il10, Il17a and Tgfb1 in the colon, ileum, Peyer’s patches and mesenteric lymph nodes of 10-week-old male (a) and female (b) mice treated with vancomycin (grey bars) and controls (open bars). Five animals per group were analyzed. Expression of Foxp3, Ifng, Il17a and Il10 was normalized to that of Cd3, while expression of Tgfb1 was normalized to Hprt expression. Results are expressed as fold of the mean expression observed in untreated mice. Only changes in expression levels outside the grey zone indicates changes in expression >2-fold increase or decrease in vancomycin-treated mice (black bars) as compared to controls (open bars). Significant differences are indicated as follows: * p<0.05; ** p<0.01.

Mentions: The expression of the following genes was analyzed by quantitative PCR in the colon, ileum, Peyer’s patches and mesenteric lymph nodes: Foxp3, Ifng, Il10, Il17a and Tgfb1 (Fig 6a and 6b). The salient result was a major reduction in the expression of Il17a in the colon, the ileum, Peyer’s patches and mesenteric lymph nodes of male and female vancomycin-treated mice (all differences were statistically significant except those in the ileum that were borderline (p = 0.0635 in males and p = 0.0593 in females)). Reduced expression of Ifng was observed only in the ileum of treated mice; the difference was statistically significant only in females. An increase in the expression of Foxp3 was found in vancomycin-treated mice as compared to untreated mice. This increase was statistically significant in the Peyer’s patches of both male and female treated mice, in the ileum of treated females and the in colon of treated males. An increase in Il10 expression was observed in Peyer’s patches of treated mice which, however, was significant only in males.


Antibiotics in early life alter the gut microbiome and increase disease incidence in a spontaneous mouse model of autoimmune insulin-dependent diabetes.

Candon S, Perez-Arroyo A, Marquet C, Valette F, Foray AP, Pelletier B, Milani C, Ventura M, Bach JF, Chatenoud L - PLoS ONE (2015)

Analysis of gene expression in the gut.Relative expression of Foxp3, Ifng, Il10, Il17a and Tgfb1 in the colon, ileum, Peyer’s patches and mesenteric lymph nodes of 10-week-old male (a) and female (b) mice treated with vancomycin (grey bars) and controls (open bars). Five animals per group were analyzed. Expression of Foxp3, Ifng, Il17a and Il10 was normalized to that of Cd3, while expression of Tgfb1 was normalized to Hprt expression. Results are expressed as fold of the mean expression observed in untreated mice. Only changes in expression levels outside the grey zone indicates changes in expression >2-fold increase or decrease in vancomycin-treated mice (black bars) as compared to controls (open bars). Significant differences are indicated as follows: * p<0.05; ** p<0.01.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4430542&req=5

pone.0125448.g006: Analysis of gene expression in the gut.Relative expression of Foxp3, Ifng, Il10, Il17a and Tgfb1 in the colon, ileum, Peyer’s patches and mesenteric lymph nodes of 10-week-old male (a) and female (b) mice treated with vancomycin (grey bars) and controls (open bars). Five animals per group were analyzed. Expression of Foxp3, Ifng, Il17a and Il10 was normalized to that of Cd3, while expression of Tgfb1 was normalized to Hprt expression. Results are expressed as fold of the mean expression observed in untreated mice. Only changes in expression levels outside the grey zone indicates changes in expression >2-fold increase or decrease in vancomycin-treated mice (black bars) as compared to controls (open bars). Significant differences are indicated as follows: * p<0.05; ** p<0.01.
Mentions: The expression of the following genes was analyzed by quantitative PCR in the colon, ileum, Peyer’s patches and mesenteric lymph nodes: Foxp3, Ifng, Il10, Il17a and Tgfb1 (Fig 6a and 6b). The salient result was a major reduction in the expression of Il17a in the colon, the ileum, Peyer’s patches and mesenteric lymph nodes of male and female vancomycin-treated mice (all differences were statistically significant except those in the ileum that were borderline (p = 0.0635 in males and p = 0.0593 in females)). Reduced expression of Ifng was observed only in the ileum of treated mice; the difference was statistically significant only in females. An increase in the expression of Foxp3 was found in vancomycin-treated mice as compared to untreated mice. This increase was statistically significant in the Peyer’s patches of both male and female treated mice, in the ileum of treated females and the in colon of treated males. An increase in Il10 expression was observed in Peyer’s patches of treated mice which, however, was significant only in males.

Bottom Line: In NOD females a slight yet not significant trend towards an increase in disease incidence was observed.Administration of the antibiotic mixture resulted in near complete ablation of the gut microbiota.These results show that a directed even partial ablation of the gut microbiota, as induced by vancomycin, significantly increases type 1 diabetes incidence in male NOD mice thus prompting for caution in the use of antibiotics in pregnant women and newborns.

View Article: PubMed Central - PubMed

Affiliation: Université Paris Descartes, Sorbonne Paris Cité, F-75475, Paris, France; INSERM U1151, Hôpital Necker-Enfants Malades, Paris, France; CNRS UMR 8253, Hôpital Necker-Enfants Malades, Paris, France.

ABSTRACT
Insulin-dependent or type 1 diabetes is a prototypic autoimmune disease whose incidence steadily increased over the past decades in industrialized countries. Recent evidence suggests the importance of the gut microbiota to explain this trend. Here, non-obese diabetic (NOD) mice that spontaneously develop autoimmune type 1 diabetes were treated with different antibiotics to explore the influence of a targeted intestinal dysbiosis in the progression of the disease. A mixture of wide spectrum antibiotics (i.e. streptomycin, colistin and ampicillin) or vancomycin alone were administered orally from the moment of conception, treating breeding pairs, and during the postnatal and adult life until the end of follow-up at 40 weeks. Diabetes incidence significantly and similarly increased in male mice following treatment with these two antibiotic regimens. In NOD females a slight yet not significant trend towards an increase in disease incidence was observed. Changes in gut microbiota composition were assessed by sequencing the V3 region of bacterial 16S rRNA genes. Administration of the antibiotic mixture resulted in near complete ablation of the gut microbiota. Vancomycin treatment led to increased Escherichia, Lactobacillus and Sutterella genera and decreased members of the Clostridiales order and Lachnospiraceae, Prevotellaceae and Rikenellaceae families, as compared to control mice. Massive elimination of IL-17-producing cells, both CD4+TCRαβ+ and TCRγδ+ T cells was observed in the lamina propria of the ileum and the colon of vancomycin-treated mice. These results show that a directed even partial ablation of the gut microbiota, as induced by vancomycin, significantly increases type 1 diabetes incidence in male NOD mice thus prompting for caution in the use of antibiotics in pregnant women and newborns.

No MeSH data available.


Related in: MedlinePlus