Unbalance of intestinal microbiota in atopic children

Marco Candela, Simone Rampelli, Silvia Turroni, Marco Severgnini, Clarissa Consolandi, Gianluca De Bellis, Riccardo Masetti, Giampaolo Ricci, Andrea Pession, Patrizia Brigidi, Marco Candela, Simone Rampelli, Silvia Turroni, Marco Severgnini, Clarissa Consolandi, Gianluca De Bellis, Riccardo Masetti, Giampaolo Ricci, Andrea Pession, Patrizia Brigidi

Abstract

Background: Playing a strategic role in the host immune function, the intestinal microbiota has been recently hypothesized to be involved in the etiology of atopy. In order to investigate the gastrointestinal microbial ecology of atopic disease, here we performed a pilot comparative molecular analysis of the faecal microbiota in atopic children and healthy controls.

Results: Nineteen atopic children and 12 healthy controls aged 4-14 years were enrolled. Stools were collected and the faecal microbiota was characterized by means of the already developed phylogenetic microarray platform, HTF-Microbi.Array, and quantitative PCR. The intestinal microbiota of atopic children showed a significant depletion in members of the Clostridium cluster IV, Faecalibacterium prausnitzii, Akkermansia muciniphila and a corresponding increase of the relative abundance of Enterobacteriaceae.

Conclusion: Depleted in key immunomodulatory symbionts, the atopy-associated microbiota can represent an inflammogenic microbial consortium which can contribute to the severity of the disease. Our data open the way to the therapeutic manipulation of the intestinal microbiota in the treatment of atopy by means of pharmaceutical probiotics.

Figures

Figure 1
Figure 1
Analysis of the HTF-Microbi.Array fluorescence signals. A: PCA of the HTF-Microbi.Array fluorescence signals. Atopy or health status were considered as dummy environmental variables (green triangles) and indicated as atopic and control, respectively. Atopic subjects and healthy controls are indicated by gray circles and red squares, respectively. Second and third ordination axes are plotted showing 6.4% and 3.3% of the total variability in the dataset, respectively. B: Comparison of the HTF-Microbi.Array probe fluorescence signals between atopics and controls. Only probes showing a different trend between the two groups (P < 0.3) are shown.
Figure 2
Figure 2
Relative contribution of the principal intestinal microbial groups in the faecal microbiota of atopics and controls. For each HTF-Microbi.Array probe, the relative fluorescence contribution was calculated as percentage of the total fluorescence. Sub-probes were excluded. Data represent the mean of the probe relative fluorescence contribution in atopics (n = 19) and controls (n = 12). P values derive from a two-sided t-test.
Figure 3
Figure 3
Spearman rank correlation between total IgE level and the abundance ofL. casei et rel.andClostridiumcluster IX in the stools from a subset of 10 atopic children.

References

    1. Romagnani S. Regulatory T cells: which role in the pathogenesis and treatment of allergic disorders? Allergy. 2006;61:3–14.
    1. Ngoc PL, Gold DR, Tzianabos AO, Weiss ST, Celedón JC. Cytokines, allergy, and asthma. Curr Opin Allergy Clin Immunol. 2005;5:161–166. doi: 10.1097/01.all.0000162309.97480.45.
    1. Penders J, Stobberingh EE, van den Brandt PA, Thijs C. The role of the intestinal microbiota in the development of atopic disorders. Allergy. 2007;62:1223–1236. doi: 10.1111/j.1398-9995.2007.01462.x.
    1. Ehlers S, Kaufmann SH. Participants of the 99(th) Dahlem Conference. Infection, inflammation, and chronic diseases: consequences of a modern lifestyle. Trends Immunol. 2010;31:184–190. doi: 10.1016/j.it.2010.02.003.
    1. Rautava S, Ruuskanen O, Ouwehand A, Salminen S, Isolauri E. The hygiene hypothesis of atopic disease–an extended version. J Pediatr Gastroenterol Nutr. 2004;38:378–388. doi: 10.1097/00005176-200404000-00004.
    1. De Filippo C, Cavalieri D, Di Paola M, Ramazzotti M, Poullet JB, Massart S, Collini S, Pieraccini G, Lionetti P. Impact of diet in shaping gut microbiota revealed by a comparative study in children from Europe and rural Africa. Proc Natl Acad Sci U S A. 2010;107:14691–14696. doi: 10.1073/pnas.1005963107.
    1. Kau AL, Ahern PP, Griffin NW, Goodman AL, Gordon JI. Human nutrition, the gut microbiome and the immune system. Nature. 2011;474:327–336. doi: 10.1038/nature10213.
    1. Lee YK, Mazmanian SK. Has the microbiota played a critical role in the evolution of the adaptive immune system? Science. 2010;330:1768–1773. doi: 10.1126/science.1195568.
    1. Egert M, de Graaf AA, Smidt H, de Vos WM, Venema K. Beyond diversity: functional microbiomics of the human colon. Trends Microbiol. 2006;14:86–91. doi: 10.1016/j.tim.2005.12.007.
    1. Mazmanian SK, Round JL, Kasper DL. A microbial symbiosis factor prevents intestinal inflammatory disease. Nature. 2008;453:620–625. doi: 10.1038/nature07008.
    1. Gaboriau-Routhiau V, Rakotobe S, Lécuyer E, Mulder I, Lan A, Bridonneau C, Rochet V, Pisi A, De Paepe M, Brandi G, Eberl G, Snel J, Kelly D, Cerf-Bensussan N. The key role of segmented filamentous bacteria in the coordinated maturation of gut helper T cell responses. Immunity. 2009;31:677–689. doi: 10.1016/j.immuni.2009.08.020.
    1. Neish AS. Microbes in gastrointestinal health and disease. Gastroenterology. 2009;136:65–80. doi: 10.1053/j.gastro.2008.10.080.
    1. Maslowski KM, Mackay CR. Diet, gut microbiota and immune responses. Nat Immunol. 2011;12:5–9.
    1. Penders J, Thijs C, van den Brandt PA, Kummeling I, Snijders B, Stelma F, Adams H, van Ree R, Stobberingh EE. Gut microbiota composition and development of atopic manifestations in infancy: the KOALA Birth Cohort Study. Gut. 2007;56:661–667. doi: 10.1136/gut.2006.100164.
    1. Kalliomäki M, Kirjavainen P, Eerola E, Kero P, Salminen S, Isolauri E. Distinct patterns of neonatal gut microflora in infants in whom atopy was and was not developing. J Allergy Clin Immunol. 2001;107:129–134. doi: 10.1067/mai.2001.111237.
    1. Vael C, Desager K. The importance of the development of the intestinal microbiota in infancy. Curr Opin Pediatr. 2009;21:794–800. doi: 10.1097/MOP.0b013e328332351b.
    1. Murray CS, Tannock GW, Simon MA, Harmsen HJ, Welling GW, Custovic A, Woodcock A. Fecal microbiota in sensitized wheezy and non-sensitized non-wheezy children: a nested case–control study. Clin Exp Allergy. 2005;35:741–745. doi: 10.1111/j.1365-2222.2005.02259.x.
    1. Penders J, Stobberingh EE, Thijs C, Adams H, Vink C, van Ree R, van den Brandt PA. Molecular fingerprinting of the intestinal microbiota of infants in whom atopic eczema was or was not developing. Clin Exp Allergy. 2006;36:1602–1608. doi: 10.1111/j.1365-2222.2006.02599.x.
    1. Bisgaard H, Li N, Bonnelykke K, Chawes BL, Skov T, Paludan-Müller G, Stokholm J, Smith B, Krogfelt KA. Reduced diversity of the intestinal microbiota during infancy is associated with increased risk of allergic disease at school age. J Allergy Clin Immunol. 2011;128:646–652. doi: 10.1016/j.jaci.2011.04.060.
    1. Abrahamsson TR, Jakobsson HE, Andersson AF, Björkstén B, Engstrand L, Jenmalm MC. Low diversity of the gut microbiota in infants with atopic eczema. J Allergy Clin Immunol. 2012;129:434–440. doi: 10.1016/j.jaci.2011.10.025.
    1. Blaser MJ, Falkow S. What are the consequences of the disappearing human microbiota? Nat Rev Microbiol. 2009;7:887–894. doi: 10.1038/nrmicro2245.
    1. Dominguez-Bello MG, Blaser MJ, Ley RE, Knight R. Development of the human gastrointestinal microbiota and insights from high-throughput sequencing. Gastroenterology. 2011;140:1713–1719. doi: 10.1053/j.gastro.2011.02.011.
    1. Palmer C, Bik EM, DiGiulio DB, Relman DA, Brown PO. Development of the human infant intestinal microbiota. PLoS Biol. 2007;5:e177. doi: 10.1371/journal.pbio.0050177.
    1. Candela M, Consolandi C, Severgnini M, Biagi E, Castiglioni B, Vitali B, De Bellis G, Brigidi P. High taxonomic level fingerprint of the human intestinal microbiota by ligase detection reaction–universal array approach. BMC Microbiol. 2010;19:116.
    1. Rajilić-Stojanović M, Smidt H, de Vos WM. Diversity of the human gastrointestinal tract microbiota revisited. Environ Microbiol. 2007;9:2125–2136. doi: 10.1111/j.1462-2920.2007.01369.x.
    1. Dreborg S, Frew A. Position Paper EAACI: allergen standardization and skin tests. Allergy. 1993;48:49–82.
    1. Cavagni G, Capra T, Suzzani L, Nori G, Derin FM, Dusi R, Bernasconi S. Valori di IgE in bambini sani, non atopici. Folia Allergol Immunol Clin. 1980;27:273.
    1. Castiglioni B, Rizzi E, Frosini A, Sivonen K, Rajaniemi P, Rantala A, Mugnai MA, Ventura S, Wilmotte A, Boutte C, Grubisic S, Balthasart P, Consolandi C, Bordoni R, Mezzelani A, Battaglia C, De Bellis G. Development of a universal microarray based on the ligation detection reaction and 16 S rrna gene polymorphism to target diversity of cyanobacteria. Appl Environ Microbiol. 2004;70:7161–7172. doi: 10.1128/AEM.70.12.7161-7172.2004.
    1. Consolandi C, Severgnini M, Castiglioni B, Bordoni R, Frosini A, Battaglia C, Rossi Bernardi L, De Bellis G. A structured chitosan-based platform for biomolecule attachment to solid surfaces: application to DNA microarray preparation. Bioconjug Chem. 2006;17:371–377. doi: 10.1021/bc050285a.
    1. Leps J, Smilauer P. Multivariate analysis of ecological data using CANOCO. Cambridge University Press, Cambridge; 2003.
    1. Collado MC, Derrien M, Isolauri E, de Vos WM, Salminem S. Intestinal integrity and Akkermansia muciniphila, a mucin-degrading member of the intestinal microbiota present in infants, adults and the elderly. Appl Environ Microbiol. 2007;23:7767–7770.
    1. Bartosch S, Fite A, Macfarlane GT, McMurdo MET. Characterization of bacterial communities in feces from healthy elderly volunteers and hospitalized elderly patients by using Real-Time PCR and effects of antibiotic treatment on the fecal microbiota. Appl Environ Microbiol. 2004;6:3575–3581.
    1. Van Dyke MI, McCarthy AJ. Molecular biological detection and characterization of Clostridium populations in municipal landfill sites. Appl Environ Microbiol. 2002;4:2049–2053.
    1. Kok RG, de Waal A, Schut F, Welling GW, Weenk G, Hellingwerf KJ. Specific detection and analysis of a probiotic Bifidobacterium strain in infant feces. Appl Environ Microbiol. 1996;62:3668–3672.
    1. Walter J, Hertel C, Tannock GW, Lis CM, Munro K, Hammes WP. Detection of Lactobacillus, Pediococcus, Leuconostoc and Weissella species in human feces by using group-specific PCR primers and denaturing gradient gel electrophoresis. Appl Environ Microbiol. 2001;67:2578–2585. doi: 10.1128/AEM.67.6.2578-2585.2001.
    1. Stsepetova J, Sepp E, Julge K, Vaughan E, Mikelsaar M, de Vos WM. Molecularly assessed shifts of Bifidobacterium ssp. and less diverse microbial communities are characteristic of 5-year-old allergic children. FEMS Immunol Med Microbiol. 2007;51:260–269. doi: 10.1111/j.1574-695X.2007.00306.x.
    1. Hong PY, Lee BW, Aw M, Shek LP, Yap GC, Chua KY, Liu WT. Comparative analysis of fecal microbiota in infants with and without eczema. PLoS One. 2010;5:e9964. doi: 10.1371/journal.pone.0009964.
    1. Penders J, Thijs C, Mommers M, Stobberingh EE, Dompeling E, Reijmerink NE, van den Brandt PA, Kerkhof M, Koppelman GH, Postma DS. Intestinal lactobacilli and the DC-SIGN gene for their recognition by dendritic cells play a role in the aetiology of allergic manifestations. Microbiology. 2010;156:3298–3305. doi: 10.1099/mic.0.042069-0.
    1. Ogawa T, Hashikawa S, Asai Y, Sakamoto H, Yasuda K, Makimura Y. A new synbiotic, Lactobacillus casei subsp. casei together with dextran, reduces murine and human allergic reaction. FEMS Immunol Med Microbiol. 2006;46:400–409. doi: 10.1111/j.1574-695X.2006.00046.x.
    1. Schiffer C, Lalanne AI, Cassard L, Mancardi DA, Malbec O, Bruhns P, Dif F, Daëron M. A strain of Lactobacillus casei inhibits the effector phase of immune inflammation. J Immunol. 2011;187:2646–2655. doi: 10.4049/jimmunol.1002415.
    1. Chow J, Mazmanian SK. A pathobiont of the microbiota balances host colonization and intestinal inflammation. Cell Host Microbe. 2010;7:265–276. doi: 10.1016/j.chom.2010.03.004.
    1. Atarashi K, Tanoue T, Shima T, Imaoka A, Kuwahara T, Momose Y, Cheng G, Yamasaki S, Saito T, Ohba Y, Taniguchi T, Takeda K, Hori S, Ivanov II, Umesaki Y, Itoh K, Honda K. Induction of colonic regulatory T cells by indigenous Clostridium species. Science. 2011;331:337–341. doi: 10.1126/science.1198469.
    1. Sokol H, Pigneur B, Watterlot L, Lakhdari O, Bermúdez-Humarán LG, Gratadoux JJ, Blugeon S, Bridonneau C, Furet JP, Corthier G, Grangette C, Vasquez N, Pochart P, Trugnan G, Thomas G, Blottière HM, Doré J, Marteau P, Seksik P, Langella P. Faecalibacterium prausnitzii is an anti-inflammatory commensal bacterium identified by gut microbiota analysis of Crohn disease patients. Proc Natl Acad Sci U S A. 2008;105:16731–16736. doi: 10.1073/pnas.0804812105.
    1. Png CW, Lindén SK, Gilshenan KS, Zoetendal EG, McSweeney CS, Sly LI, McGuckin MA, Florin TH. Mucolytic bacteria with increased prevalence in IBD mucosa augment in vitro utilization of mucin by other bacteria. Am J Gastroenterol. 2010;105:2420–2428. doi: 10.1038/ajg.2010.281.
    1. Lupp C, Robertson ML, Wickham ME, Sekirov I, Champion OL, Gaynor EC, Finlay BB. Host-mediated inflammation disrupts the intestinal microbiota and promotes the overgrowth of Enterobacteriaceae. Cell Host Microbe. 2007;2:204. doi: 10.1016/j.chom.2007.08.002.
    1. Frank DN, St Amand AL, Feldman RA, Boedeker EC, Harpaz N, Pace NR. Molecular-phylogenetic characterization of microbial community imbalances in human inflammatory bowel diseases. Proc Natl Acad Sci U S A. 2007;104:13780–13785. doi: 10.1073/pnas.0706625104.
    1. Sokol H, Seksik P, Furet JP, Firmesse O, Nion-Larmurier I, Beaugerie L, Cosnes J, Corthier G, Marteau P, Doré J. Low counts of Faecalibacterium prausnitzii in colitis microbiota. Inflamm Bowel Dis. 2009;15:1183–1189. doi: 10.1002/ibd.20903.
    1. Schwiertz A, Jacobi M, Frick JS, Richter M, Rusch K, Köhler H. Microbiota in pediatric inflammatory bowel disease. J Pediatr. 2010;157:240–244. doi: 10.1016/j.jpeds.2010.02.046.

Source: PubMed

Спонсоры и соавторы

Заболевания

Медикаментозные вмешательства

Подписаться