CoChat
Sign Up
Cited by 295Gut microbiota and healthGastrointestinal motility and disordersAutism Spectrum Disorder Research

Distinct Microbiome-Neuroimmune Signatures Correlate With Functional Abdominal Pain in Children With Autism Spectrum Disorder

Open access
Luna, Ruth Ann, Oezguen, Numan, Balderas, Miriam, Venkatachalam, Alamelu, Runge, Jessica K., Versalovic, James, Veenstra-VanderWeele, Jeremy, Anderson, George M. +2 more·2017·Cellular and Molecular Gastroenterology and Hepatology
DOI 10.1016/j.jcmgh.2016.11.008PMC PMC5331780
Open this paper in CoChat — free accountOpen PDF

Abstract

BACKGROUND & AIMS: Emerging data on the gut microbiome in autism spectrum disorder (ASD) suggest that altered host-microbe interactions may contribute to disease symptoms. Although gut microbial communities in children with ASD are reported to differ from individuals with neurotypical development, it is not known whether these bacteria induce pathogenic neuroimmune signals. METHODS: Because commensal clostridia interactions with the intestinal mucosa can regulate disease-associated cytokine and serotonergic pathways in animal models, we evaluated whether microbiome-neuroimmune profiles (from rectal biopsy specimens and blood) differed in ASD children with functional gastrointestinal disorders (ASD-FGID, n = 14) compared with neurotypical (NT) children with FGID (NT-FGID, n = 15) and without abdominal pain (NT, n = 6). Microbial 16S ribosomal DNA community signatures, cytokines, and serotonergic metabolites were quantified and correlated with gastrointestinal symptoms. RESULTS: , were evident. Stratification by abdominal pain showed multiple organisms in ASD-FGID that correlated significantly with cytokines (interleukin [IL]6, IL1, IL17A, and interferon-γ). Group comparisons showed that IL6 and tryptophan release by mucosal biopsy specimens was highest in ASD children with abdominal pain, whereas serotonergic metabolites generally were increased in children with FGIDs. Furthermore, proinflammatory cytokines correlated significantly with several Clostridiales previously reported to associate with ASD, as did tryptophan and serotonin. CONCLUSIONS: Our findings identify distinctive mucosal microbial signatures in ASD children with FGID that correlate with cytokine and tryptophan homeostasis. Future studies are needed to establish whether these disease-associated Clostridiales species confer early pathogenic signals in children with ASD and FGID.

AI snapshot

Written from the paper’s full text.

TL;DR

In children with autism spectrum disorder and functional GI disorders, rectal mucosal microbiome analysis showed increased Clostridiales and decreased Dorea, Blautia, and Sutterella, with correlations to cytokines and tryptophan/serotonin pathways. These signatures were especially linked to functional abdominal pain, but the findings are correlative and require confirmation.

Key findings

  • In ASD-FGID (n = 14) compared with NT-FGID (n = 15) and NT (n = 6), increased Clostridiales included Clostridium lituseburense (P = .002), Lachnoclostridium bolteae (P = .017), Lachnoclostridium hathewayi (P = .030), Clostridium aldenense (P = .038), and Flavonifractor plautii (P = .038); decreases included Dorea formicigenerans (P = .006), Blautia luti (P = .020), and Sutterella species (P = .025).
  • Stratification by abdominal pain showed associations with C aldenense (P = .004), C lituseburense (P = .003), F plautii (P = .007), Clostridium disporicum (P = .049), Clostridium tertium (P = .045), and Turicibacter sanguinis (P = .033) across pain groups; within ASD-FGID, pain vs no pain was associated with C aldenense (P = .033) and F plautii (P = .042).
  • Decreased tryptophan levels in ASD-FGID supernatants were significant vs NT-FGID (P = .006) and NT (P = .009); 5-HIAA was increased in ASD-FGID vs NT (P = .009) and associated with abdominal pain (P = .039). IL6 was highest in ASD-FGID, with the trend exaggerated in the abdominal pain subset.
  • Multiple cytokines correlated with ASD-FGID-associated organisms, including C lituseburense with IL1RA, IL12p70, IL1α, IL5, IL6, IP10, MIP-1α, MIP-1β, and VEGF, and C disporicum/C tertium with IFN-γ, IL12p70, IL17A, IL5, IL6, IP-10, MIP-1α, MIP-1β, and VEGF.

Methods

  • Cross-sectional study of children aged 3–18 years undergoing lower endoscopy at Nationwide Children’s Hospital; final analysis included 35 children: ASD-FGID (n = 14), NT-FGID (n = 15), and NT (n = 6). ASD diagnosis was Autism Diagnostic Observation Schedule–confirmed; non-ASD participants had Social Responsiveness Scale scores below 51; QPGS-RIII determined FGID status.
  • Rectal biopsy specimens and blood were collected; mucosal microbiome was characterized by 16S ribosomal DNA V1V3 and V4 sequencing, cytokines by a 38-plex assay, and serotonin, 5-HIAA, and tryptophan by high-performance liquid chromatography.
  • Analyses included principal component analysis, ANOVA and Welch t tests, Wilcoxon rank-sum tests, and Pearson correlations with Benjamini–Hochberg correction for multiple testing.

Limitations

  • The paper states that findings are correlative and that future studies are needed to establish whether disease-associated Clostridiales confer early pathogenic signals; it also states the sample size did not allow focus on an ASD subgroup.
  • Evident from the text: the final sample was small (n = 35), especially the NT group (n = 6); the ASD-FGID group was all male, and only 3 females were recruited overall, all in the NT-FGID group, limiting sex and generalizability considerations.
  • Evident from the text: the cross-sectional design cannot establish causality or directionality between microbiome, neuroimmune markers, and abdominal pain.

Model-written summary. Check it against the paper before relying on it.

Integrity signals

Registry

Crossref · 10.1016/j.jcmgh.2016.11.008

Cites

65 of 65 references verified0 citation errors · 0 not checkable

Statistics

Nothing to recompute.

Claims the paper makes

0 of 67 cited sources checked · 0 supported · 0 flagged

  • Findingown data

    In comparison between ASD-FGID group with pain and ASD-FGID group with no pain, Tyzzerella species (P = .045) and Parasutterella excrementihominis (P = .037) were significantly associated with the ASD-FGID group with pain.

    “Tyzzerella species (P = .045) and Parasutterella excrementihominis (P = .037) also emerged as significantly associated with the ASD-FGID group with pain compared with the ASD-FGID group with no pain.”
    Quote in the paperSubstantial claim
  • Findingown data

    IL6 release by mucosal biopsy specimens was highest in ASD children with abdominal pain.

    “Group comparisons showed that IL6 and tryptophan release by mucosal biopsy specimens was highest in ASD children with abdominal pain”
    Quote in the paperSubstantial claim
  • Findingown data

    Tryptophan release by mucosal biopsy specimens was highest in ASD children with abdominal pain.

    “Group comparisons showed that IL6 and tryptophan release by mucosal biopsy specimens was highest in ASD children with abdominal pain”
    Quote in the paperSubstantial claim
  • Findingown data

    Decreases in F plautii (P = .039), Bacteroides eggerthii (P = .024), Bacteroides uniformis (P = .040), F prausnitzii (P = .013), and Clostridium clariflavum (P = .031) were associated with functional constipation.

    “decreases in several organisms were associated with functional constipation including F plautii (P = .039), Bacteroides eggerthii (P = .024), Bacteroides uniformis (P = .040), F prausnitzii (P = .013), and Clostridium clariflavum (P = .031).”
    Quote in the paperSubstantial claim
  • Findingown data

    Serotonergic metabolites generally were increased in children with FGIDs.

    “serotonergic metabolites generally were increased in children with FGIDs.”
    Quote in the paperNot a substantial claim
  • May combine two claims. 19
  • Classified as probably not a substantial claim. 34
  • The quote only partly states this claim. 3

  • May combine two claims.

    Findingown data

    In ASD-FGID compared with NT-FGID and NT groups, there was an increase in Clostridiales OTUs identified as Clostridium lituseburense (P = .002), Lachnoclostridium bolteae (P = .017), Lachnoclostridium hathewayi (P = .030), Clostridium aldenense (P = .038), and Flavonifractor plautii (P = .038).

    “an increase in Clostridiales, specifically sequences identified as Clostridium lituseburense (P = .002), Lachnoclostridium bolteae (P = .017), Lachnoclostridium hathewayi (P = .030), Clostridium aldenense (P = .038), and Flavonifractor plautii (P = .038), was observed in the ASD-FGID group compared with the NT-FGID and NT groups”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    There was overlap between the NT-FGID and NT groups, with the NT group forming a distinct cluster within the larger NT-FGID group.

    “there was overlap between the NT-FGID and NT groups, with the NT group forming a distinct cluster within the larger NT-FGID group.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    Aerophagia was associated with decreases in B luti (P = .003), Bifidobacterium adolescentis (P = .019), Eubacterium ventriosum (P = .045), Anoxystipes fissicatena (P = .024), Coprococcus comes (P = .042), Eubacterium ramulus (P = .006), and Phascolarctobacterium faecium (P = .049).

    “as well as with a decrease in a variety of other organisms including B luti (P = .003), Bifidobacterium adolescentis (P = .019), Eubacterium ventriosum (P = .045), Anoxystipes fissicatena (P = .024), Coprococcus comes (P = .042), Eubacterium ramulus (P = .006), and Phascolarctobacterium faecium (P = .049).”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Findingown data

    No difference was identified based on sex when comparing all neurotypical participants.

    “No difference was identified based on sex when comparing all neurotypical participants.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    Decreases in Akkermansia muciniphila (P = .038), Coprococcus catus (P = .007), Odoribacter splanchnicus (P = .050), Clostridium lactatifermentans (P = .033), and Ruminococcus lactaris (P = .037) were associated with the presence of abdominal migraine.

    “Decreases in Akkermansia muciniphila (P = .038), Coprococcus catus (P = .007), Odoribacter splanchnicus (P = .050), Clostridium lactatifermentans (P = .033), and Ruminococcus lactaris (P = .037) were associated with the presence of abdominal migraine.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    In the 13-18 years group, there were increases in OTUs most closely resembling Alistipes putredinis (P = .003) and Clostridium perfringens (P = .029), and a decrease in Parabacteriodes distasonis (P = .036).

    “increases in OTUs most closely resembling Alistipes putredinis (P = .003) and Clostridium perfringens (P = .029) in the 13–18 years group, a decrease in Parabacteriodes distasonis (P = .036) in the 13–18 years group”
    Quote in the paperNot a substantial claim

  • The quote only partly states this claim.

    Findingown data

    Sutterella species correlated positively with IL12 p70.

    “Organisms decreased in abundance in the ASD-FGID cohort that correlated positively with T helper 1 regulatory cytokines included D formicigenerans (IL12 p70) and Sutterella species (IL12 p70).”
    Quote in the paperNot a substantial claim

  • The quote only partly states this claim.

    Findingown data

    Tryptophan and serotonin correlated significantly with several Clostridiales previously reported to associate with ASD.

    “as did tryptophan and serotonin.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    IL6 was correlated significantly with C disporicum and C tertium, which were associated with ASD and abdominal pain.

    “IL6 also was correlated significantly with 2 Clostridium species (C disporicum and C tertium) that were associated with ASD and abdominal pain.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Although no significant group differences or trends were seen for IL17A, IL17A correlated with Clostridia species that were enriched in ASD-FGID patients.

    “Although no significant group differences or trends were seen for IL17A, this cytokine correlated with Clostridia species that were enriched in ASD-FGID patients.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    IFN-γ was increased in ASD and correlated with C disporicum and C tertium.

    “Similarly, IFN-γ has been noted as increased in individuals with ASD,13, 16, 17, 61 and IFN-γ was increased in ASD and correlated with C disporicum and C tertium.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Premisecitation

    Altered microbiome-gut-brain signals have been reported in ASD and may contribute independently toward clinical symptoms by changing microbiome composition, tryptophan-serotonin imbalance, and immunologic pathways.

    “Altered microbiome–gut–brain signals also have been reported in ASD, and may contribute independently toward clinical symptoms by changing microbiome composition, tryptophan–serotonin imbalance, and immunologic pathways.11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21”
    Quote in the paperSubstantial claim0 of 3 sources checked
    • cites [11] Daly, 2014, Response inhibition and serotonin in autism: a functional MRI study using acute tryptophan depletion, Brain source

      Not checked in the free run.

    • cites [12] Croonenberghs, 2002, Increased serum albumin, gamma globulin, immunoglobulin IgG, and IgG2 and IgG4 in autism, Psychol Med source

      Not checked in the free run.

    • cites [13] Croonenberghs, 2002, Activation of the inflammatory response system in autism, Neuropsychobiology source

      Not checked in the free run.

  • Classified as probably not a substantial claim.

    Premisecitation

    Differences in the types and composition of various bacterial species in children with ASD have been reported.

    “Differences in the types and composition of various bacterial species in children with ASD have been reported.22, 23, 24, 25, 26, 27”
    Quote in the paperSubstantial claim0 of 3 sources checked
    • cites [22] Finegold, 2010, Pyrosequencing study of fecal microflora of autistic and control children, Anaerobe source

      Not checked in the free run.

    • cites [23] Williams, 2011, Impaired carbohydrate digestion and transport and mucosal dysbiosis in the intestines of children with autism and gastrointestinal disturbances, PLoS One source

      Not checked in the free run.

    • cites [24] Gondalia, 2012, Molecular characterisation of gastrointestinal microbiota of children with autism (with and without gastrointestinal dysfunction) and their neurotypical siblings, Autism Res source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Mucosa-associated microbes preferentially regulate host homeostasis, including induction of T-cell immune responses and maintaining serotonin biosynthesis from dietary tryptophan in the mucosa.

    “mucosa-associated microbes preferentially regulate host homeostasis, as is evident in the induction of T-cell immune responses, as well as maintaining serotonin biosynthesis from dietary tryptophan in the mucosa.28, 29, 30, 31, 32, 33”
    Quote in the paperSubstantial claim0 of 3 sources checked
    • cites [28] Wang, 2014, Increased proportions of Bifidobacterium and the Lactobacillus group and loss of butyrate-producing bacteria in inflammatory bowel disease, J Clin Microbiol source

      Not checked in the free run.

    • cites [29] Li, 2015, Diversity of duodenal and rectal microbiota in biopsy tissues and luminal contents in healthy volunteers, J Microbiol Biotechnol source

      Not checked in the free run.

    • cites [30] Reigstad, 2015, Gut microbes promote colonic serotonin production through an effect of short-chain fatty acids on enterochromaffin cells, FASEB J source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Study design and conductnone given

    The research protocol was approved by the Nationwide Children's Institutional Review Board, and written consent was obtained from parents of the participants.

    “The research protocol was approved by the Nationwide Children’s Institutional Review Board, and written consent was obtained from parents of the participants.”
    Quote in the paperNot a substantial claim

  • Classified as probably not a substantial claim.

    Study design and conductnone given

    ASD participants were required to have an Autism Diagnostic Observation Schedule-confirmed diagnosis.

    “ASD participants were required to have an Autism Diagnostic Observation Schedule–confirmed diagnosis.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Most previous studies conducted their analysis at higher taxonomic levels, with little published data at the genus or species (OTU) level.

    “Stool-based microbiome characterization in several cohorts identified specific genera that differed in ASD vs control subjects, but most previous studies conducted their analysis at higher taxonomic levels, with little published data at the genus or species (ie, OTU) level.22, 25, 26, 27”
    Quote in the paperNot a substantial claim0 of 4 sources checked
    • cites [22] Finegold, 2010, Pyrosequencing study of fecal microflora of autistic and control children, Anaerobe source

      Not checked in the free run.

    • cites [25] Kang, 2013, Reduced incidence of Prevotella and other fermenters in intestinal microflora of autistic children, PLoS One source

      Not checked in the free run.

    • cites [26] De Angelis, 2013, Fecal microbiota and metabolome of children with autism and pervasive developmental disorder not otherwise specified, PLoS One source

      Not checked in the free run.

    • cites [27] Wang, 2013, Increased abundance of Sutterella spp. and Ruminococcus torques in feces of children with autism spectrum disorder, Mol Autism source

      Not checked in the free run.

  • May combine two claims.

    Interpretationown data

    Our findings identify distinctive mucosal microbial signatures in ASD children with FGID that correlate with cytokine and tryptophan homeostasis.

    “Our findings identify distinctive mucosal microbial signatures in ASD children with FGID that correlate with cytokine and tryptophan homeostasis.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Interpretationown data

    This analysis shows functional associations that distinguish both the clinical group and GI symptoms.

    “This analysis shows functional associations that distinguish both the clinical group and GI symptoms.”
    Quote in the paperSubstantial claim

  • Classified as probably not a substantial claim.

    Study design and conductreasoning

    The Social Responsiveness Scale score less than 51 was used as cut-off for appropriate non-ASD participants.

    “If the Social Responsiveness Scale score was less than the standardized cut-off value of 51, the participant was considered appropriate for the study.”
    Quote in the paperSubstantial claim

  • Classified as probably not a substantial claim.

    Study design and conductnone given

    Blood and biopsy specimens were collected only from participants with a normal-appearing colonoscopy as determined by the pediatric GI specialist.

    “Blood and biopsy specimens were collected only from participants with a normal-appearing colonoscopy as determined by the pediatric GI specialist.”
    Quote in the paperSubstantial claim

  • Classified as probably not a substantial claim.

    Study design and conductnone given

    P values were corrected for multiple testing with the Benjamini-Hochberg method to control for false-discovery rate.

    “The P values were corrected for multiple testing with the Benjamini–Hochberg method to control for false-discovery rate.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing). The quote only partly states this claim.

    Study design and conductreasoning

    A total of 35 children were included in the final analysis after excluding 3 with abnormal colonoscopy.

    “Therefore, 35 children were included in the final analysis and are described in Table 1.”
    Quote in the paperNot a substantial claim

  • Classified as probably not a substantial claim.

    Study design and conductnone given

    Non-ASD participants were screened with the Social Responsiveness Scale to ensure they did not show behaviors in the range of those often displayed in children with ASD.

    “Non-ASD participants were screened with the Social Responsiveness Scale to ensure they did not show behaviors in the range of those that often are displayed in children with ASD.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Study design and conductnone given

    For missing cytokine measurements, a value of 10% of the lowest measurement for that cytokine was inserted.

    “For missing cytokine measurements, a value of 10% of the lowest measurement for that cytokine was inserted.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    A number of biomarkers that could connect to microbial profiles have been identified in ASD, including alterations in serotonin levels, IL1β, and IL6.

    “A number of biomarkers that could connect to microbial profiles also have been identified in ASD, including alterations in serotonin levels, IL1β, and IL6.35, 44, 45, 46, 47”
    Quote in the paperNot a substantial claim0 of 5 sources checked
    • cites [35] Choi, 2016, The maternal interleukin-17a pathway in mice promotes autism-like phenotypes in offspring, Science source

      Not checked in the free run.

    • cites [44] Boccuto, 2013, Decreased tryptophan metabolism in patients with autism spectrum disorders, Mol Autism source

      Not checked in the free run.

    • cites [45] Muller, 2016, The serotonin system in autism spectrum disorder: from biomarker to animal models, Neuroscience source

      Not checked in the free run.

    • cites [46] Masi, 2015, Cytokine aberrations in autism spectrum disorder: a systematic review and meta-analysis, Mol Psychiatry source

      Not checked in the free run.

    • cites [47] Gabriele, 2014, Blood serotonin levels in autism spectrum disorder: a systematic review and meta-analysis, Eur Neuropsychopharmacol source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Early stool-based culture and real-time polymerase chain reaction-based studies reported an increased abundance of Clostridium species in the GI tract of children with autism.

    “Early stool-based culture and real-time polymerase chain reaction–based studies reported an increased abundance of Clostridium species in the GI tract of children with autism,48, 49, 50”
    Quote in the paperNot a substantial claim0 of 3 sources checked
    • cites [48] Finegold, 2002, Gastrointestinal microflora studies in late-onset autism, Clin Infect Dis source

      Not checked in the free run.

    • cites [49] Song, 2004, Real-time PCR quantitation of clostridia in feces of autistic children, Appl Environ Microbiol source

      Not checked in the free run.

    • cites [50] Parracho, 2005, Differences between the gut microflora of children with autistic spectrum disorders and that of healthy children, J Med Microbiol source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Study design and conductown data

    The cohort included 3 female and 32 male children, assigned to ASD-FGID (n = 14), NT-FGID (n = 15), and NT (n = 6).

    “The cohort included 3 female and 32 male children and, after review of all data, subjects were assigned to the following groups: ASD-FGID (n = 14), NT-FGID (n = 15), and NT (n = 6).”
    Quote in the paperNot a substantial claim

  • Classified as probably not a substantial claim.

    Study design and conductnone given

    The QPGS-RIII is a 71-item parent report instrument used in clinical research to assess chronicity and severity of GI symptoms and characterize FGID criteria.

    “The QPGS-RIII is a 71-item parent report instrument that is used in clinical research to assess the chronicity and severity of GI symptoms and helps to characterize which particular FGID criteria of GI symptoms are met”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Stool-based microbiome characterization in several cohorts identified specific genera that differed in ASD vs control subjects.

    “Stool-based microbiome characterization in several cohorts identified specific genera that differed in ASD vs control subjects, but most previous studies conducted their analysis at higher taxonomic levels, with little published data at the genus or species (ie, OTU) level.22, 25, 26, 27”
    Quote in the paperNot a substantial claim0 of 4 sources checked
    • cites [22] Finegold, 2010, Pyrosequencing study of fecal microflora of autistic and control children, Anaerobe source

      Not checked in the free run.

    • cites [25] Kang, 2013, Reduced incidence of Prevotella and other fermenters in intestinal microflora of autistic children, PLoS One source

      Not checked in the free run.

    • cites [26] De Angelis, 2013, Fecal microbiota and metabolome of children with autism and pervasive developmental disorder not otherwise specified, PLoS One source

      Not checked in the free run.

    • cites [27] Wang, 2013, Increased abundance of Sutterella spp. and Ruminococcus torques in feces of children with autism spectrum disorder, Mol Autism source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    IFN-γ has been noted as increased in individuals with ASD.

    “Similarly, IFN-γ has been noted as increased in individuals with ASD,13, 16, 17, 61 and IFN-γ was increased in ASD and correlated with C disporicum and C tertium.”
    Quote in the paperNot a substantial claim0 of 4 sources checked
    • cites [13] Croonenberghs, 2002, Activation of the inflammatory response system in autism, Neuropsychobiology source

      Not checked in the free run.

    • cites [16] Li, 2009, Elevated immune response in the brain of autistic patients, J Neuroimmunol source

      Not checked in the free run.

    • cites [17] Chez, 2007, Elevation of tumor necrosis factor-alpha in cerebrospinal fluid of autistic children, Pediatr Neurol source

      Not checked in the free run.

    • cites [61] Singh, 1996, Plasma increase of interleukin-12 and interferon-gamma. Pathological significance in autism, J Neuroimmunol source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    A recent publication discussed the significant increase in C bolteae in otherwise healthy individuals after antibiotic administration.

    “A recent publication also discussed the significant increase in C bolteae in otherwise healthy individuals after antibiotic administration,52”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [52] Raymond, 2016, The initial state of the human gut microbiome determines its reshaping by antibiotics, ISME J source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    An increase in inflammatory cytokines has been reported previously in ASD.

    “An increase in inflammatory cytokines has been reported previously in ASD, and increased cytokines have been reported in association with a regressive autism phenotype with significant communication challenges and aberrant behaviors.56, 57”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites [56] Ashwood, 2011, Associations of impaired behaviors with elevated plasma chemokines in autism spectrum disorders, J Neuroimmunol source

      Not checked in the free run.

    • cites [57] Ashwood, 2011, Elevated plasma cytokines in autism spectrum disorders provide evidence of immune dysfunction and are associated with impaired behavioral outcome, Brain Behav Immun source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Turicibacter species was reported previously in pediatric patients with IBS who failed to respond to a low FODMAP diet.

    “In addition, Turicibacter species was associated with abdominal pain in the ASD-FGID cohort and was reported previously in pediatric patients with IBS who failed to respond to a low fermentable oligosaccharides, disaccharides, monosaccharides, and polyols diet.55”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [55] Chumpitazi, 2015, Randomised clinical trial: gut microbiome biomarkers are associated with clinical response to a low FODMAP diet in children with the irritable bowel syndrome, Aliment Pharmacol Ther source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Prior research in pediatric IBS identified increases in γ-proteobacteria, specifically Haemophilus parainfluenzae, and a Ruminococcus-like organism.

    “Prior research in pediatric IBS identified increases in γ-proteobacteria, specifically Haemophilus parainfluenzae, and a Ruminococcus-like organism.54”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [54] Saulnier, 2011, Gastrointestinal microbiome signatures of pediatric patients with irritable bowel syndrome, Gastroenterology source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisecitation

    Finegold et al originally described increased Clostridium in ASD in 2002, and ultimately identified a novel species, Clostridium (Lachnoclostridium) bolteae, associated with ASD, specifically in those with GI symptoms.

    “Finegold et al48 originally described increased Clostridium in ASD in 2002, and ultimately identified a novel species, Clostridium (Lachnoclostridium) bolteae, associated with ASD, specifically in those with GI symptoms.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [48] Finegold, 2002, Gastrointestinal microflora studies in late-onset autism, Clin Infect Dis source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    In the single tissue-based pediatric ASD study, an increase in Clostridiales, especially Lachnospiraceae and Ruminococcaceae, was observed.

    “Although the analysis focused mainly on higher taxonomic levels, an increase in Clostridiales, especially Lachnospiraceaea and Ruminococcaceae, was observed.23”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [23] Williams, 2011, Impaired carbohydrate digestion and transport and mucosal dysbiosis in the intestines of children with autism and gastrointestinal disturbances, PLoS One source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Increased cytokines have been reported in association with a regressive autism phenotype with significant communication challenges and aberrant behaviors.

    “An increase in inflammatory cytokines has been reported previously in ASD, and increased cytokines have been reported in association with a regressive autism phenotype with significant communication challenges and aberrant behaviors.56, 57”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites [56] Ashwood, 2011, Associations of impaired behaviors with elevated plasma chemokines in autism spectrum disorders, J Neuroimmunol source

      Not checked in the free run.

    • cites [57] Ashwood, 2011, Elevated plasma cytokines in autism spectrum disorders provide evidence of immune dysfunction and are associated with impaired behavioral outcome, Brain Behav Immun source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Increased IL6 has been seen in clinical cohorts of ASD.

    “Furthermore, increased IL6 has been seen in clinical cohorts of ASD.13, 14, 17, 60”
    Quote in the paperNot a substantial claim0 of 4 sources checked
    • cites [13] Croonenberghs, 2002, Activation of the inflammatory response system in autism, Neuropsychobiology source

      Not checked in the free run.

    • cites [14] Jyonouchi, 2001, Proinflammatory and regulatory cytokine production associated with innate and adaptive immune responses in children with autism spectrum disorders and developmental regression, J Neuroimmunol source

      Not checked in the free run.

    • cites [17] Chez, 2007, Elevation of tumor necrosis factor-alpha in cerebrospinal fluid of autistic children, Pediatr Neurol source

      Not checked in the free run.

    • cites [60] Hsiao, 2011, Activation of the maternal immune system induces endocrine changes in the placenta via IL-6, Brain Behav Immun source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    A single tissue-based study in pediatric ASD has been published with a total sample size of 22 subjects (ASD, n = 15; NT, n = 7) and a limited age range of 3-5 years.

    “A single tissue-based study in pediatric ASD has been published with a total sample size of 22 subjects (ASD, n = 15; NT, n = 7) and a limited age range of 3–5 years.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [23] Williams, 2011, Impaired carbohydrate digestion and transport and mucosal dysbiosis in the intestines of children with autism and gastrointestinal disturbances, PLoS One source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Interpretationcitation

    The identification of clostridial species in this study aligns with previous autism studies that have identified microbiome alterations.

    “Overall, our identification of clostridial species aligns with those previous autism studies that have identified microbiome alterations.”
    Quote in the paperNot a substantial claim

  • Classified as probably not a substantial claim. May combine two claims.

    Premisecitation

    IL6 and IL17A are both implicated in maternal immune activation models that result in autism-like and schizophrenia-like phenotypes in rodents.

    “IL6 and IL17A are both implicated in the maternal immune activation models that result in autism-like and schizophrenia-like phenotypes in rodents.58, 59”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites [58] Hsiao, 2012, Modeling an autism risk factor in mice leads to permanent immune dysregulation, Proc Natl Acad Sci U S A source

      Not checked in the free run.

    • cites [59] Wu, 2015, The interaction between maternal immune activation and alpha 7 nicotinic acetylcholine receptor in regulating behaviors in the offspring, Brain Behav Immun source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Real-time polymerase chain reaction analysis targeting C bolteae confirmed the increased abundance in ASD.

    “Real-time polymerase chain reaction analysis targeting C bolteae confirmed the increased abundance in ASD,49”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [49] Song, 2004, Real-time PCR quantitation of clostridia in feces of autistic children, Appl Environ Microbiol source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Long-standing findings have indicated decreased levels of the serotonin metabolite melatonin in a substantial subpopulation of children with ASD.

    “Long-standing findings have indicated increased whole-blood 5-HT levels in a subset of approximately 30% of children with autism spectrum disorder,44, 45, 46, 47 as well as decreased levels of the serotonin metabolite, melatonin, in a substantial subpopulation.62”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [62] Pagan, 2014, The serotonin-N-acetylserotonin-melatonin pathway as a biomarker for autism spectrum disorders, Transl Psychiatry source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Long-standing findings have indicated increased whole-blood 5-HT levels in a subset of approximately 30% of children with autism spectrum disorder.

    “Long-standing findings have indicated increased whole-blood 5-HT levels in a subset of approximately 30% of children with autism spectrum disorder,44, 45, 46, 47 as well as decreased levels of the serotonin metabolite, melatonin, in a substantial subpopulation.62”
    Quote in the paperNot a substantial claim0 of 4 sources checked
    • cites [44] Boccuto, 2013, Decreased tryptophan metabolism in patients with autism spectrum disorders, Mol Autism source

      Not checked in the free run.

    • cites [45] Muller, 2016, The serotonin system in autism spectrum disorder: from biomarker to animal models, Neuroscience source

      Not checked in the free run.

    • cites [46] Masi, 2015, Cytokine aberrations in autism spectrum disorder: a systematic review and meta-analysis, Mol Psychiatry source

      Not checked in the free run.

    • cites [47] Gabriele, 2014, Blood serotonin levels in autism spectrum disorder: a systematic review and meta-analysis, Eur Neuropsychopharmacol source

      Not checked in the free run.

  • Classified as probably not a substantial claim.

    Interpretationnone given

    GI phenotypes in ASD are highly variable.

    “In summary, GI phenotypes in ASD are highly variable, and heterogeneity of the autism population can mask significant signals related to GI symptomology.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Whole-blood 5-HT levels correlate with lower GI symptoms in ASD.

    “Interestingly, whole-blood 5-HT levels correlate with lower GI symptoms in ASD.63”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [63] Marler, 2016, Brief report: whole blood serotonin levels and gastrointestinal symptoms in autism spectrum disorder, J Autism Dev Disord source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Interpretationnone given

    Heterogeneity of the autism population can mask significant signals related to GI symptomology.

    “In summary, GI phenotypes in ASD are highly variable, and heterogeneity of the autism population can mask significant signals related to GI symptomology.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Interpretationown data

    These data suggest the presence of a specific microbiome profile in ASD with the identification of organisms previously strongly associated within similar cohorts, albeit in stool-based studies.

    “These data suggest the presence of a specific microbiome profile in ASD with the identification of organisms previously strongly associated within similar cohorts, albeit in stool-based studies.”
    Quote in the paperSubstantial claim
    • Rests on an assumption: The rectal mucosal microbiome and cytokine measurements reflect the gut mucosal environment relevant to functional abdominal pain in ASD.

  • May combine two claims.

    Interpretationown data

    These ASD-related organisms interact with the intestinal mucosa and are associated with altered neuroimmune signaling.

    “We advance this concept by showing that these ASD-related organisms interact with the intestinal mucosa and are associated with altered neuroimmune signaling.”
    Quote in the paperSubstantial claim
    • Rests on an assumption: The rectal mucosal microbiome and cytokine measurements reflect the gut mucosal environment relevant to functional abdominal pain in ASD.

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    A recent study showed that an ASD-associated serotonin transporter variant leads to altered gut development and function.

    “Furthermore, a recent study showed that an ASD-associated serotonin transporter variant leads to altered gut development and function.64”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [64] Margolis, 2016, Serotonin transporter variant drives preventable gastrointestinal abnormalities in development and function, J Clin Invest source

      Not checked in the free run.

51 of 51

  • The paper relies on this without support.

    It is not known whether gut microbial communities in children with ASD induce pathogenic neuroimmune signals.

    “it is not known whether these bacteria induce pathogenic neuroimmune signals.”

  • The paper relies on this without support.

    The etiology of FGIDs is poorly understood in children with neurotypical development and in ASD.

    “The etiology of FGIDs is poorly understood in children with neurotypical development, let alone in ASD.”

  • The paper relies on this without support.

    It is unethical to perform an endoscopy on children with ASD without a clinical indication.

    “Because it is unethical to perform an endoscopy on children with ASD without a clinical indication, the only cohort that can be captured is generally ASD with FGID.”

  • The paper relies on this without support.

    Most studies have focused on stool specimens because it is difficult to obtain adequate numbers of GI mucosal biopsy specimens from children with ASD.

    “Most studies have focused on stool specimens because it is difficult to obtain adequate numbers of GI mucosal biopsy specimens from children with ASD.”

  • The paper relies on this without support.

    The rectal mucosal microbiome and cytokine measurements reflect the gut mucosal environment relevant to functional abdominal pain in ASD.

    “Here, we present distinct microbiome and cytokine measures in the rectal mucosa of children with ASD and confirmed FGIDs compared with neurotypical children both with and without FGIDs.”

  • The paper relies on this without support.

    The NT-FGID and NT comparison groups are adequate to isolate ASD-FGID-specific microbiome and cytokine signatures.

    “compared with neurotypical children both with and without FGIDs.”

  • The paper relies on this without support.

    The presence of abdominal pain can be validly classified by separating the cohort based on abdominal pain.

    “Upon separation of the cohort (all 3 groups) based on abdominal pain, increases in MCP-1 (P = .033) and eotaxin (P = .028) were associated significantly with the presence of abdominal pain.”
7 of 7

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

Each contradiction between two passages listed here was confirmed by an independent check.

  • Conflicting descriptions of the correlation test

    Minor

    The statistical methods say Pearson correlations were calculated for microbial OTUs, cytokines, and serotonin. The results instead say the cytokine and serotonergic correlations with microbial OTUs were performed using a Wilcoxon rank-sum test.

1 of 1

Cites

Flagged first, then by citations

Cited by

Sorted by citations

Related

Authors
Luna, R. A., Oezguen, N., Balderas, M., Venkatachalam, A., Runge, J. K., Versalovic, J., Veenstra-VanderWeele, J., Anderson, G. M., Savidge, T., & Williams, K. C.
Affiliations
  • Baylor College of Medicine (US)
  • Columbia University (US)
  • Yale University (US)
  • Nationwide Children's Hospital (US)
Corresponding author
Luna, Ruth Ann
ORCID
Venue
Cellular and Molecular Gastroenterology and Hepatology
Publisher
Elsevier BV
Published
2017-03
Volume
3
Issue
2
Pages
218-230
ISSN
2352-345X (ISSN-L)
DOI
10.1016/j.jcmgh.2016.11.008
PMC
PMC5331780
OpenAlex
W2559803799
Semantic Scholar
2e6891b214761db0ae748fe816fb08c592130921
Cited by
295
References
65
Citations by year
2026: 12 · 2025: 21 · 2024: 33 · 2023: 30 · 2022: 48 · 2021: 41 · 2020: 39 · 2019: 40 · 2018: 22 · 2017: 8
FWCI
12.12
Type
journal article
Language
en
Open access
Licence
Locations
  • Cellular and Molecular Gastroenterology and Hepatology (publishedVersion, cc-by-nc-nd)
  • Europe PMC (PubMed Central) (publishedVersion, cc-by-nc-nd)
  • PubMed Central (publishedVersion, cc-by-nc-nd)
  • PubMed (publishedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Funders
  • Health Resources and Services Administration
  • Government of South Australia
  • U.S. Department of Health and Human Services (UA3 MC11054)
  • National Institute of Allergy and Infectious Diseases (RO1AI100914, U01 AI124290-01)
  • Autism Speaks (9455)
  • National Institute of Diabetes and Digestive and Kidney Diseases (DK56338, UH2 DK093990, UH3 DK083990)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Gut microbiota and health
  • Health Sciences › Medicine › Gastroenterology › Gastrointestinal motility and disorders
  • Life Sciences › Neuroscience › Cognitive Neuroscience › Autism Spectrum Disorder Research
Keywords
autism spectrum disorder, functional abdominal pain, functional gastrointestinal disorders, gut microbiota, neuroimmune signaling, inflammatory cytokines, interleukin-6, tryptophan metabolism, serotonin, Clostridiales, intestinal mucosa, children

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Claims read by deepseek-v4.1-flash under definition version 3 (extract-v3) on 2026-10-09.

Classified by jev-1.13-20260917 (gold-guideline-v0.2) on 2026-10-09.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review