A real-time PCR assay for accurate quantification of the individual members of the Altered Schaedler Flora microbiota in gnotobiotic mice

Gomes-Neto, João Carlos; Mantz, Sara; Held, Kyler; Sinha, Rohita; Muñoz, Rafael R. Segura; Schmaltz, Robert; Benson, Andrew K.; Walter, Jens; Ramer‐Tait, Amanda E.

doi:10.1016/j.mimet.2017.02.003

Cited by 45 publications

(63 citation statements)

References 43 publications

(78 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…ASF mice are a reliable model to study how the addition of a specific bacterial strain to a stable community influences immunological and metabolic pathways or affects disease outcomes (Gomez de Aguero et al, 2016; Shen et al, 2015)(Gomes-Neto et al, 2017). Having observed that fluconazole-induced fungal dysbiosis exerts persistent impacts on systemic immunity and AAD in mycobiota-sufficient specific pathogen-free (SPF) mice, without directly targeting either commensal bacteria or the host, we asked whether a supplementation with intestinal fungi can recapitulate the disease phenotype in MyF-ASF mice.…”

Section: Resultsmentioning

confidence: 99%

Response to Fungal Dysbiosis by Gut-Resident CX3CR1+ Mononuclear Phagocytes Aggravates Allergic Airway Disease

Leonardi

Semon

et al. 2018

Cell Host & Microbe

Self Cite

105

104

View full text Add to dashboard Cite

Summary Sensing of the gut microbiota, including fungi, regulates mucosal immunity. Whether fungal sensing in the gut can influence immunity at other body sites is unknown. Here we show that fluconazole-induced gut fungal dysbiosis has persistent effects on allergic airway disease in a house dust mite challenge model. Mice with a defined community of bacteria, but lacking intestinal fungi were not susceptible to fluconazole-induced dysbiosis, while colonization with a fungal mixture recapitulated the detrimental effects. Gut resident mononuclear phagocytes (MNPs) expressing the fractalkine receptor CX3CR1 were essential for the effect of gut fungal dysbiosis on peripheral immunity. Depletion of CX3CR1+ MNPs or selective inhibition of Syk signaling downstream of fungal sensing in these cells ameliorated lung allergy. These results indicate that disruption of intestinal fungal communities can have persistent effects on peripheral immunity and aggravate disease severity through fungal sensing by gut resident CX3CR1+ MNPs.

show abstract

Section: Resultsmentioning

confidence: 99%

Response to Fungal Dysbiosis by Gut-Resident CX3CR1+ Mononuclear Phagocytes Aggravates Allergic Airway Disease

Leonardi

Semon

et al. 2018

Cell Host & Microbe

Self Cite

105

104

View full text Add to dashboard Cite

show abstract

“…Male and female C3H/HeN mice (8-10 weeks old) harboring the Altered Schaedler Flora (ASF) microbiota from birth were bred and maintained under gnotobiotic conditions at the University of Nebraska-Lincoln Gnotobiotic Mouse Facility as previously described. 47,48 Members of ASF community include: ASF 356, Clostridium sp. ; ASF 360, Lactobacillus intestinalis; ASF361, Lactobacillus murinus; ASF 457, Mucispirillum schaedleri; ASF 492, Eubacterium plexicaudatum; ASF 500, Pseudoflavonifractor sp.…”

Section: Micementioning

confidence: 99%

“…49,50 Prior to inoculation with E. coli, ASF-bearing mice were transferred from flexible film isolators to a positive pressure, individually-ventilated caging system and maintained as previously reported. 48 Each mouse received 1 x 10 8 E. coli CFU in 200 µL of LB broth via oral gavage. Successful E. coli colonization was verified by collecting fecal samples 10 days post-inoculation and plating on Eosin Methylene Blue (EMB) agar plates (Difco).…”

Section: Micementioning

confidence: 99%

See 1 more Smart Citation

Establishing the phenotypic basis of adherent-invasiveEscherichia coli(AIEC) pathogenicity in intestinal inflammation

Kittana

Gomes-Neto

Heck

et al. 2019

Preprint

Self Cite

View full text Add to dashboard Cite

Background & Aims: Adherent-invasive Escherichia coli (AIEC) are enriched in ileal Crohn's disease patients and implicated in disease etiology. However, AIEC pathogenesis is poorly understood, and it is unclear if the expansion of these organisms contributes to inflammatory bowel disease (IBD). Questions also remain as to what extent the various in vitro phenotypes used to classify AIEC are pathologically relevant. Methods: We utilized a combination of in vitro phenotyping and a murine model of intestinal inflammation to systematically relate AIEC phenotypes to pathogenicity for 30 mucosa-associated humanderived E. coli strains. In vitro assays used included survival/replication in and TNF-α production by J774 macrophages as well as invasion/replication in Caco2 intestinal epithelial cells.Results: AIEC do not form a phenotypic group that is clearly separated from non-AIEC.However, E. coli strains displaying in vitro AIEC phenotypes caused, on average, more severe intestinal inflammation. Survival/replication of strains in J774 and Caco2 cells were positively correlated with disease in vivo, while adherence to Caco2 cells and TNF-α production by J774 cells were not. Importantly, co-colonization with adherent non-AIEC strains ameliorated AIECmediated disease. Conclusion: Our findings do not support the existence of an AIEC pathovar that can be clearly separated from commensal E. coli. However, intracellular survival/replication phenotypes do contribute to murine intestinal inflammation, suggesting that the AIEC overgrowth observed in human IBD makes a causal contribution to disease. The ability to differentiate pathologically-relevant AIEC phenotypes from those that are not provides an important foundation for developing strategies to predict, diagnose and treat human IBD through characterizing and modulating patient E. coli populations.

show abstract

“…In order to generate mice that harbor the Altered Schaedler Flora (ASF) microbiota, germ-free (GF) mice were gavaged with cecal contents of ASF mice. Colonization was determined by qPCR as previously reported (26). Sulfur amino acids (Saa) diets were formulated based on previous literature (27,28) to represent edge cases of Saa consumption,…”

mentioning

confidence: 99%

Diet Post-translationally Modifies The Gut Microbial Proteome To Modulate Renal Function

Lobel

Cao

Glickman

et al. 2020

Preprint

View full text Add to dashboard Cite

One Sentence Summary:We found that diet post-translationally modifies the gut microbiota proteome to modulate kidney function. ABSTRACT:We identify a novel mechanism linking diet, gut microbial metabolism, and renal function. We found that a sulfur amino acid-based dietary intervention post-translationally modifies a microbial enzyme, blunting its uremic toxin-producing activity and alleviating chronic kidney disease (CKD) in a preclinical model. We also define a heretofore unknown role for the post-translational modification Ssulfhydration within the gut microbiome. This study provides a framework for understanding how diet can tune microbiota function via protein post-translational modification without altering microbial community composition to support healthy host physiology beyond the gut and specifically how a dietary modification can inhibit tryptophanase activity to ameliorate CKD progression. Main Text:Chronic kidney disease (CKD) affects nearly 850 million people worldwide (1). Although dietary modification is a cornerstone of CKD treatment, the mechanistic roles of diet-microbiota interactions in CKD pathogenesis and treatment have been under-explored. While many dietmicrobiome studies have focused on the effects of dietary fiber, fat and carbohydrates (2), less is known about the specific effects of dietary protein and amino acids, although 5-10% of dietary amino acids reach the colon where most gut bacterial metabolism occurs (3). In humans, increasing dietary protein increases gut bacterial production of hydrogen sulfide (H 2 S), indole, and indoxyl sulfate (4, 5).Indole and indoxyl sulfate are uremic toxins; and H 2 S has diverse physiological functions, some of which are mediated by the post-translational modification S-sulfhydration (6, 7). While a vast number of studies have been performed in mammalian systems, the physiological roles of H 2 S in regulating gut bacterial function within a host are understudied. Additionally, whether there are bona fide opportunities to improve CKD by manipulating diet-microbiota interactions remain unclear.Given the knowledge gaps around dietary protein, gut microbial metabolism and H 2 S, and to address the role of gut microbial metabolism and diet in renal function; we employed a mouse model of CKD that is driven by elevated adenine (8) along with a sulfur amino acid (Saa)-based diet perturbation. We formulated isocaloric diets to represent edge cases of mouse Saa consumption, i.e.

show abstract

A real-time PCR assay for accurate quantification of the individual members of the Altered Schaedler Flora microbiota in gnotobiotic mice

Cited by 45 publications

References 43 publications

Response to Fungal Dysbiosis by Gut-Resident CX3CR1+ Mononuclear Phagocytes Aggravates Allergic Airway Disease

Response to Fungal Dysbiosis by Gut-Resident CX3CR1+ Mononuclear Phagocytes Aggravates Allergic Airway Disease

Establishing the phenotypic basis of adherent-invasiveEscherichia coli(AIEC) pathogenicity in intestinal inflammation

Diet Post-translationally Modifies The Gut Microbial Proteome To Modulate Renal Function

Contact Info

Product

Resources

About