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Growth and host interaction of mouse segmented filamentous bacteria in vitro


The gut microbiota plays a crucial role in the maturation of the intestinal mucosal immune system of its host1,2. Within the thousand bacterial species present in the intestine, the symbiont segmented filamentous bacterium (SFB) is unique in its ability to potently stimulate the post-natal maturation of the B- and T-cell compartments and induce a striking increase in the small-intestinal Th17 responses3,4,5. Unlike other commensals, SFB intimately attaches to absorptive epithelial cells in the ileum and cells overlying Peyer’s patches6,7. This colonization does not result in pathology; rather, it protects the host from pathogens4. Yet, little is known about the SFB–host interaction that underlies the important immunostimulatory properties of SFB, because SFB have resisted in vitro culturing for more than 50 years. Here we grow mouse SFB outside their host in an SFB–host cell co-culturing system. Single-celled SFB isolated from monocolonized mice undergo filamentation, segmentation, and differentiation to release viable infectious particles, the intracellular offspring, which can colonize mice to induce signature immune responses. In vitro, intracellular offspring can attach to mouse and human host cells and recruit actin. In addition, SFB can potently stimulate the upregulation of host innate defence genes, inflammatory cytokines, and chemokines. In vitro culturing thereby mimics the in vivo niche, provides new insights into SFB growth requirements and their immunostimulatory potential, and makes possible the investigation of the complex developmental stages of SFB and the detailed dissection of the unique SFB–host interaction at the cellular and molecular levels.

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Figure 1: Growth and growth requirements of SFB in vitro.
Figure 2: Differentiation of SFB from filaments to intracellular offspring during in vitro growth.
Figure 3: Viability, colonization, and immunostimulatory potential of in vitro-grown SFB.
Figure 4: SFB–host cell interaction and host response.


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We are grateful to B. Marteyn, F.-X. Campbell-Valois, and C. Parsot for discussions, M. Picard and S. Rakotobe for help with in vivo experiments, and T. Angelique for animal care. This work was supported by INSERM, Institut Pasteur, College de France, and INRA, and grants TORNADO-FP7-KBBE-2007-2A-222720, ANR-2010-BLAN1317, ERC-2009-AG-232798-HOMEOPITH, ERC-2013-AdG-339579-DECRYPT, and ERC-2013-AdG-339407-IMMUNOBIOTA and the Investissement d’Avenir ANR-10-IAHU-01 and LabEX IBEID. P.J.S. is a Howard Hughes Medical Institute Foreign Scholar.

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Authors and Affiliations



N.C.-B., V.G.-R., P.J.S., and P.S. conceived the project and discussed experiments. P.S. designed and performed all in vitro experiments. V.G.-R. and M.G. performed the in vivo challenge experiments, V.G.-R. maintained SFB mice and M.G. analysed the TC7 and R.F. the mICl2 host response. M.M.-N. processed SEM samples and took images with P.S. G.N. assisted in vitro experiments. P.S. wrote the paper and P.J.S., N.C.B. and V.G.-R. edited the manuscript.

Corresponding authors

Correspondence to Nadine Cerf-Bensussan or Philippe J. Sansonetti.

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The authors declare no competing financial interests.

Extended data figures and tables

Extended Data Figure 1 Intestinal colonization of in vitro-grown SFB and host response.

a, b, qPCR quantification (a) and SEM (b) of in vitro-grown SFB used for gavage. c, qPCR quantification of SFB in faecal samples. d, SEM of SFB attachment in vivo at 25 days after gavage of C57BL/6 mice with SFBVivo or SFBVitro. e, SEM of 21-day SFB-colonized germ-free C3H/HeN mouse ileum showing vacant attachment sites. f, Host gene expression in the ileal lamina propria in conventional or germ-free mice colonized with either SFB or E. coli for 21 days. Images and values are representatives from one of two experiments (ae) or are cumulative values from two experiments performed with a total of seven germ-free, four SFB, five E. coli, and four conventional mice (f), showing box plots of 25–75% centiles with median and minimum/maximum whiskers. c, f, Two-tailed t-test statistical analysis (P < 0.05, P < 0.01).

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Schnupf, P., Gaboriau-Routhiau, V., Gros, M. et al. Growth and host interaction of mouse segmented filamentous bacteria in vitro. Nature 520, 99–103 (2015).

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