The human body supports a thriving diversity of microbes which comprise a dynamic, ancillary, functional system that synergistically develops in lock-step with physiological development of its host. The human microbiome field has transitioned from cataloging this rich diversity to dissecting molecular mechanisms by which microbiomes influence human health. Early life microbiome development trains immune function. Thus, vertically, horizontally, and environmentally acquired microbes and their metabolites have the potential to shape developmental trajectories with life-long implications for health.
Our recent understanding that the human body supports a thriving diversity of microbial life has led to a greater appreciation of the expanded functional gene capacity of the human superorganism. Over the past several years, the field of human microbiome research has transitioned from cataloging the diversity of these microorganisms to the view that they comprise a dynamic ancillary functional system that synergistically develops across spatial and temporal gradients in parallel with physiological development and decline1,2,3,4,5. For centuries, we have observed how human health is influenced by microbes and that interactions between microbial and host cells govern infectious diseases. What is becoming more apparent is that a wide array of conditions ranging from chronic inflammatory6,7 and metabolic8,9,10 diseases to neurological disorders11,12 and cancer13 have now been associated with microbiome functional perturbations. These may occur either locally at the site of disease manifestation or at remote mucosal sites or organ systems, which stimulate metabolic and immunologic changes in the host.
Insights from germ-free mice
In 1885 Louis Pasteur speculated that gnotobiotic or germ-free (GF) animals would not be able to survive due to the extensive co-evolutionary history with microbes14. While GF animals can indeed be reared, studies using such animals have been instrumental in illustrating the crucial role of microbes in mammalian development and health. GF mice exhibit shorter lifespans, heavily enlarged caeca15, lack natural antibodies16,17, and are severely deficient in vitamin K and B1215. GF mice generated in genetically susceptible backgrounds enable investigation of relationships between host genetics and the microbiome. For example, conventional IL-10 deficient mice develop spontaneous colitis, but their GF counterparts neither develop colitis nor immune system activation18 pointing to the microbiome as a disease trigger in a system poised for inflammation. However, conventionalization of GF mice does not always rescue the observed defects. Upon experimental conventionalization in adults, previously GF mice display increased susceptibility to pathogens19. Supporting these findings, GF mice exhibit aberrant invariant natural killer T-cell function and increased morbidity in models of IBD and allergic asthma, which can only be rescued by conventionalization in the neonatal period but not in adulthood20. Thus, age-sensitive microbial programming appears critical to the establishment and training of immune function and subsequent health outcomes. Evidence that microbiomes and their products are responsible for human disease has also been provided using GF mice, in which transfer of patient microbiomes confers features of the disease phenotype on the recipient animal21,22,23,24,25.
Microbial metabolites influence physiology
A large number of extrinsic and intrinsic factors, including diet, antimicrobials and immunity influence human microbiomes, in particular the gut microbiome which houses the largest number and diversity of microbes. In turn, the bioactive products of the microbiome shape human cell function locally26,27 and at remote sites. In studies of mono-colonized GF mice, members of the intestinal microbiome were shown to strongly influence mammalian energy harvest and metabolism28,29 and produce a suite of microbial-specific metabolites in physiologically relevant concentrations, a large number of which enter the circulation30. Thus, microbial activities at one body habitat may influence physiological conditions and cell function at a remote site. For example, increased concentrations of trimethylamine N-oxide (TMAO) associate with arthrosclerosis and is dependent on gut bacterial metabolism of dietary phosphatiylcholine31,32, providing the first evidence for a diet by gut microbial interaction governing cardiovascular disease.
Development of the human microbiome
Microbiomes develop across body sites in early life, a process that adheres to the principals of ecological succession33 and shapes physiological and immunological function18. Indeed, bacteria detected at 4 days of age in human neonates correlate with community structure at 120 days of age34, indicating the importance of founder organisms in microbiological successional trajectories. Early-life microbiomes therefore may offer insights into the origins of disease and the capacity to both identify those at risk and intervene early to prevent disease development. For example, gut microbiome and metabolic dysfunction in infancy is characteristic of higher-risk for atopy and asthma development in childhood7. Moreover, specific microbial-derived metabolites found in elevated concentrations in the feces of high-risk for asthma infants promote key features of immune dysfunction characteristic of established disease7,35. For example, the oxylipin 12,13-DiHOME, elevated in the feces of high-risk babies, induces allergic inflammation in both primary immune cell and murine models35. Genes encoding bacterial epoxide hydrolases catalyzing the production of 12,13-DiHOME were enriched in the feces of high-risk babies. Introduction of these bacterial genes to the gut microbiome of mice was sufficient to increase circulating concentrations of 12,13-DiHOME and exacerbate airway allergic inflammation35. Moreover, in one month infant stool samples increasing concentrations of this oxylipin or the copy number of bacterial epoxide hydrolases capable of its production, significantly increased the risk for atopy and/or asthma development in childhood35, underscoring a role for early-life microbes and their metabolic products in disease development. More recently, both direct (electron microscopy) and indirect (molecular) evidence for the presence of a sparse, but viable, bacteria in the human fetal intestine by mid-gestation was reported in humans, and fetal bacterial strains isolated only in the presence of pregnancy hormones exhibited the ability to modulate fetal T-cell inflammatory ability36. Independently, development of antigen-experienced immune cell populations with the capacity to respond to microbial stimuli has been demonstrated as early as the second trimester of pregnancy in humans37,38,39,40 and continues post-natally. Thus vertically, horizontally, and environmentally acquired early-life microbes and their metabolites influence immune function and physiological development in a manner that shapes trajectories with life-long implications for health and an improved understanding of interactions that govern this process is critical.
Ecologically, low-abundance species and strain populations are essential reservoirs of genetic and functional diversity41. Broader understanding of primary and ancillary microbial functions amassed during the microbiome assembly process in early life and reassembly following perturbation in mature microbiomes is key to understanding features of microbiome function, stability and resilience. Critical also to this understanding is the role of microbe–microbe and microbe–host interactions which govern competitive colonization and niche specificity42,43 in the context of nutritional substrate availability. Immunological immaturity is observed in germ-free and laboratory mice, as compared to wild mice44,45,46 and humans residing on farms exhibit greater microbial functional diversity and a lower susceptibility to chronic inflammatory disease47,48. Traditional hunter-gatherer populations possess dynamic lineages of microbes which exhibit seasonal changes and include microbial species largely extinct in urban dwellers49, while lifestyle changes including settled habitation lead to depletion of disease protective microbes in nomadic populations50. Thus, progressive loss of the most flexible and responsive microbes to environmental exposures may occur at the expense of lifestyle changes and modernization. Microbial ecosystem management, involving precision nutrition and rational microbial supplementation to promote or reinstate microbial functional networks eroded by Western lifestyles and urban exposures may become increasingly important.
As we move into the next decade of human microbiome research, forward momentum in the field requires an understanding of microbial function, productivity, and interaction with the human host across spatial, temporal, and environmental gradients. Integrative analyses of parallel high-resolution cellular profiling approaches applied to longitudinally collected samples capturing microbiome function, productivity, host response and anthropologic measurements will lead to a broader appreciation of our co-evolution with microbes and the forces that shape microbe–microbe–host interactions. In addition to breadth, such studies also require depth; interrogation across scales from the ecosystem level to cellular and molecular networks that shape human biology are necessary to facilitate mechanistic insights necessary to leverage this field for precision diagnostics and interventions. Determining how to successfully re-populate depleted microbial functions and rationally and sustainably re-engineer microbiomes across a range of developmental stages, host genetic backgrounds and environmental exposures represents the next frontier in human microbiome research.
Stewart, C. J. et al. Temporal development of the gut microbiome in early childhood from the TEDDY study. Nature 562, 583–588 (2018).
Langille, M. G. et al. Microbial shifts in the aging mouse gut. Microbiome 2, 50 (2014).
Claesson, M. J. et al. Gut microbiota composition correlates with diet and health in the elderly. Nature 488, 178–184 (2012).
Cheng, J. et al. Discordant temporal development of bacterial phyla and the emergence of core in the fecal microbiota of young children. ISME J. 10, 1002–1014 (2016).
Hollister, E. B. et al. Structure and function of the healthy pre-adolescent pediatric gut microbiome. Microbiome 3, 36 (2015).
Arrieta, M.-C., Stiemsma, L. T., Amenyogbe, N., Brown, E. M. & Finlay, B. The intestinal microbiome in early life: health and disease. Front. Immunol. 5, 427 (2014).
Fujimura, K. E. et al. Neonatal gut microbiota associates with childhood multisensitized atopy and T cell differentiation. Nat. Med. 22, 1187–1191 (2016).
Turnbaugh, P. J. et al. A core gut microbiome in obese and lean twins. Nature 457, 480–484 (2009).
Le Chatelier, E. et al. Richness of human gut microbiome correlates with metabolic markers. Nature 500, 541–546 (2013).
Wang, J. et al. A metagenome-wide association study of gut microbiota in type 2 diabetes. Nature 490, 55–60 (2012).
Cryan, J. F., O’Riordan, K. J., Sandhu, K., Peterson, V. & Dinan, T. G. The gut microbiome in neurological disorders. Lancet Neurol. 19, 179–194 (2020).
Hsiao, E. Y. et al. Microbiota modulate behavioral and physiological abnormalities associated with neurodevelopmental disorders. Cell 155, 1451–1463 (2013).
Nejman, D. et al. The human tumor microbiome is composed of tumor type-specific intracellular bacteria. Science 368, 973–980 (2020).
Pasteur, L. Observations relatives à la note précédente de M. Duclaux. CR Acad. Sci. 100, 68 (1885).
Pleasants, J. R. Rearing germfree cesarean-born rats, mice, and rabbits through weaning. Ann. N. Y. Acad. Sci. 78, 116–126 (1959).
Ikari, N. S. Bactericidal antibody to Escherichia coli in germ-free mice. Nature 202, 879–881 (1964).
Bos, N. A., Meeuwsen, C. G., Wostmann, B. S., Pleasants, J. R. & Benner, R. The influence of exogenous antigenic stimulation on the specificity repertoire of background immunoglobulin-secreting cells of different isotypes. Cell. Immunol. 112, 371–380 (1988).
Sellon, R. K. et al. Resident enteric bacteria are necessary for development of spontaneous colitis and immune system activation in interleukin-10-deficient mice. Infect. Immun. 66, 5224–5231 (1998).
Hansen, A. K., Hansen, C. H. F., Krych, L. & Nielsen, D. S. Impact of the gut microbiota on rodent models of human disease. World J. Gastroenterol. 20, 17727–17736 (2014).
Olszak, T. et al. Microbial exposure during early life has persistent effects on natural killer T cell function. Science 336, 489–493 (2012).
Turnbaugh, P. J. et al. An obesity-associated gut microbiome with increased capacity for energy harvest. Nature 444, 1027–1031 (2006).
Ley, R. E., Turnbaugh, P. J., Klein, S. & Gordon, J. I. Microbial ecology: human gut microbes associated with obesity. Nature 444, 1022–1023 (2006).
Turnbaugh, P. J. et al. The effect of diet on the human gut microbiome: a metagenomic analysis in humanized gnotobiotic mice. Sci. Transl. Med. 1, 6ra14–6ra14 (2009).
Turnbaugh, P. J., Bäckhed, F., Fulton, L. & Gordon, J. I. Diet-induced obesity is linked to marked but reversible alterations in the mouse distal gut microbiome. Cell Host Microbe 3, 213–223 (2008).
Sharon, G. et al. Human gut microbiota from autism spectrum disorder promote behavioral symptoms in mice. Cell 177, 1600–1618.e17 (2019).
Scharschmidt, T. C. et al. A wave of regulatory T Cells into neonatal skin mediates tolerance to commensal microbes. Immunity 43, 1011–1021 (2015).
Hepworth, M. R. et al. Innate lymphoid cells regulate CD4+ T-cell responses to intestinal commensal bacteria. Nature 498, 113–117 (2013).
Bäckhed, F. et al. The gut microbiota as an environmental factor that regulates fat storage. Proc. Natl Acad. Sci. USA 101, 15718–15723 (2004).
Sonnenburg, J. L. et al. Glycan foraging in vivo by an intestine-adapted bacterial symbiont. Science (80-.) 307, 1955–1959 (2005).
Uchimura, Y. et al. Antibodies set boundaries limiting microbial metabolite penetration and the resultant mammalian host response. Immunity 545–559, https://doi.org/10.1016/j.immuni.2018.08.004 (2018).
Wang, Z. et al. Gut flora metabolism of phosphatidylcholine promotes cardiovascular disease. Nature 472, 57–63 (2011).
Tang, W. H. W. et al. Intestinal microbial metabolism of phosphatidylcholine and cardiovascular risk. N. Engl. J. Med 368, 1575–1584 (2013).
Gilbert, J. A. & Lynch, S. V. Community ecology as a framework for human microbiome research. Nat. Med. https://doi.org/10.1038/s41591-019-0464-9 (2019).
Eggesbø, M. et al. Development of gut microbiota in infants not exposed to medical interventions. APMIS 119, 17–35 (2011).
Levan, S. R. et al. Elevated faecal 12,13-diHOME concentration in neonates at high risk for asthma is produced by gut bacteria and impedes immune tolerance. Nat. Microbiol. https://doi.org/10.1038/s41564-019-0498-2 (2019).
Rackaityte, E. et al. Viable bacterial colonization is highly limited in the human intestine in utero. Nat. Med. 26, 599–607 (2020).
Stras, S. F. et al. Maturation of the human intestinal immune system occurs early in fetal development. Dev. Cell 51, 357–373.e5 (2019).
Halkias, J. et al. CD161 contributes to prenatal immune suppression of IFNγ-producing PLZF+ T cells. J. Clin. Invest. https://doi.org/10.1172/JCI125957 (2019).
Schreurs, R. R. C. E. et al. Human fetal TNF-α-cytokine-producing CD4+ effector memory T cells promote intestinal development and mediate inflammation early in life. Immunity 0, 1–15 (2019).
Li, N. et al. Memory CD4+ T cells are generated in the human fetal intestine. Nat. Immunol. https://doi.org/10.1038/s41590-018-0294-9 (2019).
Lynch, M. D. J. & Neufeld, J. D. Ecology and exploration of the rare biosphere. Nat. Rev. Microbiol. 13, 217–229 (2015).
Suez, J. et al. Post-antibiotic gut mucosal microbiome reconstitution is impaired by probiotics and improved by autologous FMT. Cell 174, 1406–1423.e16 (2018).
Zmora, N. et al. Personalized gut mucosal colonization resistance to empiric probiotics is associated with unique host and microbiome features. Cell 174, 1388–1405.e21 (2018).
Rosshart, S. P. et al. Wild mouse gut microbiota promotes host fitness and improves disease resistance. Cell 171, 1015–1021.e13 (2017).
Rosshart, S. P. et al. Laboratory mice born to wild mice have natural microbiota and model human immune responses. Science (80-.) 365, eaaw4361 (2019).
Beura, L. K. et al. Normalizing the environment recapitulates adult human immune traits in laboratory mice. Nature 532, 512–516 (2016).
Stein, M. M. et al. Innate immunity and asthma risk in amish and hutterite farm children. N. Engl. J. Med. 375, 411–421 (2016).
Dhakal, S. et al. Amish (rural) vs. Non-amish (urban) infant fecal microbiotas are highly diverse and their transplantation lead to differences in mucosal immune maturation in a humanized germfree piglet model. Front. Immunol. 10, 1509 (2019).
Smits, S. A. et al. Seasonal cycling in the gut microbiome of the Hadza hunter-gatherers of Tanzania. Science (80-.) 357, 802–806 (2017).
Keohane, D. M. et al. Microbiome and health implications for ethnic minorities after enforced lifestyle changes. Nat. Med. 26, 1089–1095 (2020).
E.R. was supported by NSF Graduate Research Fellowship grant no. 1650113 and by the National Institute of Allergy and Infectious Diseases (NIAID) of the NIH F31AI136336. S.V.L. was supported by NIH NIAID grant no. AI114271 and UG30D023282. The content is solely the responsibility of the authors and does not necessarily represent the official views of the NSF or the NIH.
S.V.L. is co-founder of Siolta Therapeutics Inc. and serves as both a consultant and a member of its Board of Directors.
Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
About this article
Cite this article
Rackaityte, E., Lynch, S.V. The human microbiome in the 21st century. Nat Commun 11, 5256 (2020). https://doi.org/10.1038/s41467-020-18983-8