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- Jemima Sanders
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1 doi:1.138/nture1331 A helthy volunteer (Jpnese, mle) +hu Fig. 1-c Fig. 1e Fig. 1e feces +x1 4 -re +x1 4 -re-re Chlorofrom tretment +huchlo Cecl contents +x1 4 x1 dilution (recoloniztion) x1 dilution (recoloniztion) cohoused x1 4 dilution Fig. 1e Cecl contents x1 4 dilution Fig. 1e +(x1 4 ) Fig. 1d In vitro Culture of cecl contents 31 cteril strins selection 3 cteril strins selection Fig. d,e fig. S6 +3-mix individul strin 3 strins 17 cteril strins ll 17 strins strins strins strins +-mix-a +-mix-b +-mix-c +3-mix Fig. 3,,c Fig. 3c Fig. 3c Fig. 3c Fig. 3c fig S1 Schemtic representtion of the strtegy to isolte Treg-inducing strins from the humn gut microiot. The ornge rectngle highlights ex mice where full induction of Treg cells ws oserved. 1
2 fig. S RESEARCH SUPPLEMENTARY INFORMATION +hu +huchlo cell numer hu +huchlo IL IFN-γ fig. S Treg, Th1 nd Th17 cells ccumulted in ex mice inoculted with humn microiot. IQI mice were inoculted with humn feces treted without or with chloroform (+hu nd +huchlo, respectively). (), IL-17 nd IFN-γ () expression y CD4 + cells in the colon LP were nlysed 4 weeks lter y flow cytometry. Representtive histogrms nd dot plots within the CD4 + cell popultion of individul mice re shown. Comined dt of ll mice re shown in FIg. 1-c.
3 RESEARCH Reltive undnces of OTUs (%) Bcteroidetes Others Clostridi SPF +hu +huchlo fig. S3 Assessment of microiot composition in SPF, +hu nd +huchlo mice. Pyrosequencing of 16S rrna genes ws performed on cecl contents from SPF, +hu nd +huchlo mice using 44 sequencer. Reltive undnce of OTUs (%) in the cecl cteril community in ech mouse is shown. 3
4 RESEARCH SUPPLEMENTARY INFORMATION 4 Strin strin 1184 strin3 strin strin strin7 1 strin strin9 13 strin1 143 strin strin strin13 17 strin14 1 strin strin strin strin18 14 strin strin Sequenced length (p) strin strin strin1 141 strin 143 strin3 149 strin4 146 strin 491 strin strin7 14 strin strin9 137 strin3 149 strin Close reltives Clostridium scchrogumi (T) SDG-Mt8-3D Clostridium sp Clostridium spiroforme DSM 1 Clostridium rmosum JCM 34 Clostridium rmosum DSM 14 Lchnospircee cterium 7 1 8FAA Flvonifrctor plutii ATCC 9863 Clostridium hthewy Clostridium hthewyi WAL 1868 Clostridium hthewyi DSM Clostridium scindens; VPI 178 Lchnospircee cterium 1 7FAA Bluti coccoides 8F Bluti product JCM 1471 Lchnospircee cterium 6_1_63FAA Clostridium oltee ATCC BAA 613 Clostridium oltee (T) type strin 1631 Clostridicee cterium JC13 Erysipelotrichcee cterium 44A Clostridium indolis CM971 Anerostipes ccce DSM 1466 Clostridium oltee (T); type strin: 1631 Clostridium oltee ATCC BAA 613 Clostridium hthewyi; RCGLD1 Clostridium hthewyi WAL 1868 Lchnospircee cterium 3_1_7FAA_CT1 Anerotruncus colihominis DSM 1741 Anerotruncus colihominis HKU19 Ruminococcus sp. ID8 Coprococcus comes ATCC 778 Clostridium sprgiforme DSM 1981 Clostridium lvlense CCRI-999 Clostridium sp FAA Clostridium symiosum WAL Eucterium contortum; type strin: DSM 398 Clostridium sp. D Eucterium fissicten Clostridium rmosum DSM 14 Clostridium rmosum JCM 34 Clostridium hthewy Clostridium hthewyi DSM Clostridium hthewyi RCGLD1 Clostridium hthewy Clostridium hthewyi WAL 1868 Eucterium contortum type strin DSM 398 Eucterium fissicten Clostridium D Clostridium rmosum DSM 14 Clostridium rmosum JCM 34 Oscillospircee cterium NML 6148 Oscillicter vlericigenes Clostridium sp. D Eucterium contortum; type strin: DSM 398 Clostridium scindens VPI 178 Clostridium scindens ATCC 374 Lchnospircee cterium 3 1 7FAA CT1 Clostridiles cterium 1_7_47FAA Clostridium ldenense (T) RMA 9741 Lchnospircee cterium 3 1 7FAA CT1 Clostridium hthewyi RCGLD1 Clostridium hthewyi Clostridium hthewyi WAL 1868 Lchnospircee cterium 3 1 7FAA CT1 Dtse Similrity (%) Lchnospircee cterium 3 1 7FAA CT Mtch length Similrlity with other strin strin 18 (>99%) strin 4 (>99%) strin 1 (>99%) strin 4 (>99%) strin 4 (>99%) strin 18 (>99%) strin 4 (>99%) strin 9 (>99%) 3-mix 17-mix -mixa -mixb -mixc 3-mix fig. S4 16S rrna gene nlysis for ech of the isolted 31 strins. The full-length or prt of the 16S rrna gene ws mplified y colony-pcr using 16S rrna gene-specific primer pirs. Ech mplified 16S rrna gene ws sequenced nd compred to the Riosoml Dtse Project () dtse nd genome dtse () constructed from puliclly ville genome sequences in NCBI nd HMP dtses. Close reltive species/strins, % similrity, nd the similrity to other strins re shown. Strins tht were included in the 3-mix, 17-mix, -mix-a, -mix-b, -mix-c, nd 3-mix re mrked in green. 4
5 RESEARCH +3-mix mix c 1 4 (Il1 Venus ) (Il1 Venus ) CD CD cell numer Helios IL-1 (Venus) CTLA fig. S ex mice colonised with 3-mix or 17-mix., Representtive photogrph of intestines of (left) nd +3-mix (right) mice. Note the shrunken cecum in the +3-mix mouse., Lymphocytes from colonic LP of nd +3-mix mice were nlysed for the expression of CD4, nd Helios y flow cytometry. Representtive histogrms show Helios expression y CD4 + + lymphocytes. (See lso Fig.e.) c, Colon LP lymphocytes from - or 17-mix-colonised-Il1 Venus mice were nlysed for expression of CD4,, Venus nd CTLA4 y flow cytometry. Representtive dot-plots for Venus nd CTLA4 expression y CD4 + + lymphocytes re shown.
6 RESEARCH SUPPLEMENTARY INFORMATION % + in CD4 6 4 % IL17 + in CD4 1 1 St.8 St.1 St.3 St.13 St.18 St.16 St.1 St.9 St.14 St.6 St.7 St.6 St.1 St.9 St.8 St.7 St.4 All 17 St.8 St.1 St.3 St.13 St.18 St.16 St.1 St.9 St.14 St.6 St.7 St.6 St.1 St.9 St.8 St.7 St.4 All 17 fig. S6 Treg induction cpility of ech strin. Percentges of + cells () nd IL-17 + cells () in the CD4 + cell popultion in colonic lmin propri of IQI ex mice monocolonised with ech of the 17 strins. Circles represent smples from individul nimls. Error rs, s.d. P <.1; P <. vs., s clculted y Student s t-test. 6
7 RESEARCH T-β1 (pg/ml) 1 1 Cco T84 HT9 Colo d T-β1 (pg/ml) T-β1 (pg/ml) CMT93 (mouse EC) IL-6 (pg/ml) c Tβ1 (pg/ml) 1 1 TNFα (pg/ml) HCT8 NS NS humn primry intestinl epithelil cells +ProK +Nuclese fig. S7 T-β production y epithelil cells following stimultion with cecl extrcts from mice. The cecl contents from mice nd mice were collected nd suspended in wter. The cecl wter extrcts were dded to the culture of humn intestinl epithelil cell lines ( nd c), mouse epithelil cell line (), nd humn primry intestinl epithelil cells (d). The production of T-β1 (ctive-form), IL-6 nd TNFα ws ssessed y ELISA. In pnel c, the cecl extrcts were pre-tretmed with proteinse K or nuclese. Circles represent cecl extrct from individul nimls. Error rs, s.d. P <.1; ns, not significnt, s clculted y Student s t-test. 7
8 RESEARCH SUPPLEMENTARY INFORMATION μmol/g content μmol/g content Acette..1 St.3 St.1 St.8 +-mix-c St.3 St.1 St.8 +-mix-c Propionte.3 Lctte..4 Succinte.3..1 St.3 St.1 St.8 +-mix-c St.3 St.1 St.8 +-mix-c.1.1. St.3 T-β1 (pg/ml) Isoutyrte St.1 St mix-c 3 1 St.3 Butyrte St.1 Sodium cette Sodium utyrte Sodium propionte Sodium isoutyrte SCFA mix St.8 +-mix-c fig. S8 SCFAs stimulte intestinl epithelil cells to produce T-β., The concentrtions of orgnic cids (cette, propionte, iso-utyrte, utyrte, succinte nd lctte) in the cecl contents from single strin-colonised, +-mix-c or mice (n=3 per group) were mesured y gs chromtogrphy. The concentrtions of cette, propionte, iso-utyrte nd utyrte in cecl contents from mice were higher thn those in +-mix-c mice, fr higher thn those in single strin colonized mice, nd were correlted with the cpcity to induce T-β in epithelil cell lines., The epithelil cell line HT9 ws stimulted either with. mm sodium cette, sodium propionte, sodium utyrte, sodium isoutyrte, or their mixture (SCFA mix) for 4 h. Dt re men ± SD of triplicte mesurements representtive of two independent experiments. The concentrtion of ctive-form T-β1 in the culture superntnts ws mesured y ELISA. SCFA mix induced T-β1 production t level equivlent to tht y cecl extrcts from mice. Error rs, S.D. P <.1, s clculted y the Student s t-test. 8
9 RESEARCH OT-I Teff Treg OT-II Treg or LP Treg c 1 OT-I pep OT-I cell numer (x1 4 ) OT-I pep lone Teff : Treg 1: 1 splenic CD11c+ 1:.1 1:. 1:.37 1:. OT-I pep + cecl 1 OT-I pep cecl OT-II pep or Cecl contents ( or ) or 17st in vitro Tregs ntigens OT-II Treg + OT-I pep lone OT-II Treg + OT-I pep + OT-II pep LP Treg + OT-I pep lone LP Treg + OT-I pep + OT-II pep LP Treg + OT-I pep + cecl 1 OT-I pep +17st in vitro LP Tregs CD CD OT-I T (Teff) fig. S9 Recognition of clostridil ntigens y Treg cells in the LP of mice., Schemtic representtion of cognte ntigen-driven suppression ssy., CD11c + cells were isolted from spleens of SPF C7BL/6 mice, pulsed with. μm SIINFEKL OT-I peptide lone or in comintion with either of μm ISQAVHAAHAEINEAGR OT-II peptide or utoclved cecl contents from mice, nd plted t x 1 4 /well. CD8 OT-I T cells (Teff) were dded to the CD11c + cell-seeded pltes t x 1 4 /well. Then, CD4 + CD + T cells sorted from colonic LP of mice (LP Treg) or from spleens of SPF OT-II mice (OT-II Treg) were dded to the culture t the indicted rtio of Treg to Teff cells. After 3 dys, ll cells were hrvested, stined with nti-cd4 nd nti-cd8 ntiodies, nd nlysed y flow cytometry to enumerte the numer of CD8 OT-I T cells. Depicted dt represent verge of duplictes. c, CD8 T cells from OT-I mice (Teff) nd colon LP CD4 + CD + cells from mice (Treg) were incuted with CD11c + cells pulsed with OT-I peptide lone or in comintion with either of utoclved cecl contents from mice (+ cecl) or mice ( cecl), or utoclved in vitro cultured 17 strins (+17st in vitro). Representtive dot plots t 1:.4 Teff:Treg rtio re shown here, nd summrized dt re shown in Fig. 3e. 9
10 RESEARCH SUPPLEMENTARY INFORMATION fig. S1 fig. S1 Colon Colon w w w w 3w w w 3w Colon Colon w 1. w w w 3w w w w c % + + in in CD d 6 d colon LP colon LP colon LP colon LP weeks fter inocultion weeks fter inocultion weeks weeks fter fter inocultion inocultion fig. fig. S1 S1 Frequencies Frequencies of of cells nd + cells fter colonistion with 17-mix. IQI mice were colonised with 17-mix. The + cells nd percentges of + cells + fter colonistion with cells within the CD mix. IQI mice were colonised cell popultion ( nd c), nd with 17-mix. The percentges cells within the CD4 of + cells within the CD4 + cell popultion ( nd c), nd + cells within the CD4 + + cell cell popultion popultion ( ( nd nd d) d) in in the the colon colon nd nd spleen spleen of of individul mice were nlysed y y flow flow cytometry cytometry t t the the indicted indicted weeks weeks fter fter colonistion. colonistion. Representtive Representtive histogrms nd dot plots re re shown shown in in nd nd,, nd nd summrized summrized dt dt re re shown shown in in c c nd nd d. indi- d. % + + in in CD4 CD
11 RESEARCH fig. S11 fig. S11 Colon w 1w w 3w w 1w w 4 3w Colon CD13 CD13 CD13 CD CD CD CD CD Colon Colon CD CD13 CD CD CD CD13 CD13 CD13 CD13 w 1w w 3w CD w 1w w 3w c % CD13 + in CD integrin β7 integrin β7 d c d colon 7 8 colon LP LP 7 colon LP 8 colon LP weeks weeks fter fter inocultion weeks fter inocultion % CD13 + in CD4 + + % Integrin Integrin β7 β7 + in in CD4 CD fig. S11 fig. S11 Frequencies Frequencies of CD13 of CD13 + cells + cells nd nd integrin-β7 integrin-β7 + + cells cells fter colonistion with with 17-mix. 17-mix. IQI mice were colonised with 17-mix. The percentges of CD13 IQI mice were colonised with 17-mix. The percentges of CD13 + cells ( nd c) nd Integrin-β7 cells ( nd c) nd Integrin-β7 + + cells cells ( nd d) within the CD4 ( nd d) within the CD cell + cell popultion in the colon nd spleen of individul mice were nlysed y flow cytometry t the indicted weeks fter colonistion. Representtive dot plots re shown in popultion in the colon nd spleen of individul mice were nlysed y flow cytometry t the indicted weeks fter colonistion. Representtive dot plots re shown in nd, nd summrized dt re shown in c nd d. nd, nd summrized dt re shown in c nd d. 11
12 RESEARCH SUPPLEMENTARY INFORMATION Strin Totl ses Scffolds in ssemly Totl contig length (M) Numer of predicted genes Strin_1 3,6, ,171 Strin_3 86,81, ,897 Strin_4 197,86, ,717 Strin_6 174,789, ,66 Strin_7 11,3, ,968 Strin_8 17,87, ,31 Strin_9 41,164, ,619 Strin_13 36,38, ,66 Strin_14 13,867, ,1 Strin_1 19,78, ,76 Strin_16 17,976, ,7 Strin_18 3,67, ,69 Strin_1 3,917,96 8.4,139 Strin_6 341,876, ,93 Strin_7 179,8, ,74 Strin_8 18,91, ,736 Strin_9,66, ,91 Similr genome (size) Clostridium spiroforme DSM 1 (.1 M) Flvonifrctor plutii ATCC 9863 (3.81 M) Clostridium hthewyi DSM (6.63 M) Lchnospircee cterium 6_1_63FAA (.7 M) Clostridium oltee ATCC BAA-613 (6.6 M) Erysipelotrichcee cterium 44A (4.97 M) Anerostipes ccce DSM 1466 (3.61 M) Anerotruncus colihominis DSM 1741 (3.7 M) Coprococcus comes ATCC 778 (3.4 M) Clostridium sprgiforme DSM 1981 (6. M) Clostridium symiosum WAL (.3 M) Clostridium rmosum DSM 14 (3.3 M) Clostridium sp. D (.3 M) Clostridium scindens ATCC 374 (3.6 M) Lchnospircee cterium 3_1_7FAA_CT1 (7.69 M) Clostridiles cterium 1_7_47FAA (6.1 M) Lchnospircee cterium 3_1_7FAA_CT1 (7.69 M) DDBJ BioProject ID PRJDB1 PRJDB PRJDB3 PRJDB PRJDB6 PRJDB7 PRJDB8 PRJDB3 PRJDB33 PRJDB34 PRJDB3 PRJDB36 PRJDB37 PRJDB4 PRJDB41 PRJDB4 PRJDB43 fig. S1 Genome sequencing nlysis of the 17 strins. Sequencing ws conducted using 44GS FLX Ti nd Ion PGM sequencers. Conctented sequences consisted of 6 riosoml protein genes predicted in ech genome were used for serch of similr genomes in the Genome dtse. 1
13 RESEARCH Lchnospircee_cterium 1_7FAA 1 Clostridium_scindens_ATCC_ St6 1 Dore_formicigenerns_ATCC_77 73 Clostridium_hylemone_DSM_13 Ruminococcus_lctris_ATCC_9176 Ruminococcus_gnvus_ATCC_ St1 99 Clostridium_sp_D Clostridium_nexile_DSM_ St14 3 Coprococcus_comes_ATCC_778 Ruminococcus_oeum_ATCC_ St Lchnospircee_cterium_6_1_63FAA 1 Bluti_hnsenii_DSM_83 Bryntell_formtexigens_DSM_14469 St St9 1 Lchnospircee_cterium_3_1_7FAA_CT Eucterium_rectle_ATCC_3366 Roseuri_intestinlis_L1-8 Butyrivirio_crossotus_DSM_876 Eucterium_ventriosum_ATCC_76 1 St16 Clostridium_symiosum_WAL_ St4 1 Clostridium_hthewyi_DSM_ Clostridium_scchrolyticum_WM1 1 1 St1 97 Clostridium_sprgiforme_DSM_ St8 Clostridiles_cterium_1_7_47FAA 99 Clostridium_clostridioforme 1_49FAA 99 St7 98 Clostridium_oltee_ATCC_BAA-613 Eucterium_hllii_DSM_ St9 1 Anerostipes_ccce_DSM_ St1 1 Clostridium_spiroforme_DSM_1 1 St18 99 Clostridium_rmosum_DSM_ St8 Erysipelotrichcee_cterium 44A Clostridium_thermocellum_ATCC_74 Clostridium_cellulolyticum_H1 Clostridium_hirnonis_DSM_137 Clostridium_difficile_ATCC_43 Clostridium_novyi_NT Clostridium_otulinum_D_str_1873 Clostridium_utyricum_1 Clostridium_eijerinckii_NCIMB_8 Clostridium_perfringens_ATCC_1314 Clostridium_cetoutylicum_ATCC_84 Clostridium_tetni_E88 Clostridium_kluyveri_DSM_ Selenomons_sputigen_ATCC_318 Mitsuokell_multcid_DSM_44 Acidminococcus_fermentns_DSM_731 Clostridium_leptum_DSM_ St13 Anerotruncus_colihominis_DSM_1741 Sudoligrnulum_vriile_DSM_1176 Feclicterium_prusnitzii_A-16 Lchnospircee_cterium_7_1_8FAA 1 St3 9 Flvonifrctor_plutii_ATCC_9863 Clostridi cluster XIV cluster XVIII (Erysipelotrichi) cluster III cluster XI cluster I cluster IX cluster IV fig. S13 Phylogenetic distriution of the 17 strins. The phylogenetic tree ws constructed using the MEGA v. pckge nd the neighour-joining method with ootstrp of 1 replictes. 13
14 RESEARCH SUPPLEMENTARY INFORMATION fig. S14 Helthy UC Strin1 Strin3 Strin4 Strin6 Strin7 Strin8 Strin9 Strin13 Strin14 Strin1 Strin16 Strin18 Strin1 Strin6 Strin7 Strin8 Strin9 B. frgilis NCTC 9343 F. prusnitzii A-16 F. prusnitzii M1 MH MH3 MH6 MH9 MH11 MH1 MH14 MH16 MH MH1 MH4 MH MH6 MH8 MH3 O.UC-1 O.UC-11 O.UC-1 O.UC-13 O.UC-14 O.UC-16 O.UC-17 O.UC-18 O.UC-19 O.UC- O.UC-1 O.UC- O.UC-3 O.UC-4 O.UC-4 V1.UC-1 V1.UC-13 V1.UC-14 V1.UC-1 V1.UC No. of mpped reds Strin Strin Strin Strin Strin7 1 1 Strin Strin Strin Strin14 No. of mpped reds Strin Strin Strin Strin Strin Strin Strin Strin9 fig. S14 Reltive undnce of the 17 strins in the humn fecl microiot of UC nd helthy sujects. The pulicly ville reds of 1 helthy nd UC sujects in the MetHIT dtse (only dt sets constructed with reds over 7 p length were used) were ligned to the genome of ech of the 17 strins with the cut-off vlue of the identity of >9% nd the length coverge of >9%. The numer of mpped reds ws normlised y totl red numer nd the genome size (per 1M reds per 1M of genome sequence). The het mp ws constructed using z-score normlistion of the mpping results, in which the reltive undnce of ech strin ws represented s the colour intensity of ech grid (green, low undnce; red, high undnce). The men numers of mpped reds in the helthy nd UC groups for ech of the 17 strin genomes re shown s r grphs. Error rs, SEM. P <., s clculted y the Student s t-test. 14
15 RESEARCH Dirrhe score (n=7) +PC61 (n=7) (n=7) -1 OVA+CFA (sc) 1 3 (W) OVA (orl) PC61 (ip) PC61 (ip) fig. S1 Attenution of the protective effect of 17-mix y Treg depletion in n llergen-induced dirrhoe model. BALB/c SPF dult mice were primed y sucutneous injection with 1 mg of OVA in 1 μl of CFA. One week fter priming, mice were given mg of OVA dissolved in μl of PBS y intr-gstric dministrtion three times per week. 17-mix or control PBS ws orlly dministered to mice every or 3 dys for the entire period of the experiments. SPF+17- mix group of mice were split into two sugroups, nd ech one (n=7) ws treted with or without.36 mg of nti-cd Treg-depleting ntiody (PC61) y intrperitonel injection efore the first nd fourth OVA chllenge. Dirrhoe ws monitored nd scored visully 1 h fter ech orl OVA chllenge. Error rs, s.e.m. P <.1; P <. vs.+pc61, s clculted y the Student s t-test 1
16 RESEARCH SUPPLEMENTARY INFORMATION mrna T-β1 mrna IL-1β mrna..1 IL-6 mrna.1 fig. S16 Effect of 17-mix on TNBS colitis. SPF+17mix nd SPF+cont mice were treted with TNBS., Gross morphology of the colons on dy 4 fter TNBS dministrtion., The distl intestines were collected from mice in ech group on dy 4 fter TNBS dministrtion nd exmined for expression of, T-β1, IL-1β nd IL-6 mrna y quntittive PCR. Error rs, s.d. P <.1; P <., s clculted y the Student s t-test. 16
17 RESEARCH ody weight (%) CD4 + + cell numer (x 1 4 ) week fter T cell trnsfer CD4RB hi cell trnsfer orl inocultion with SPF feces +/- 17-mix c T-β1 mrna.3..1 T-β mrna mrna IL-6 mrna TNFα mrna..1 fig. S17 Effect of 17-mix on n doptive T cell trnsfer model of colitis. To colonize 17-mix together with SPF microiot, C.B-17 SCID mice were orlly inoculted with feces from SPF mice together with or without 17-mix [ (n=) or (PBS, n=4), respectively]. One week lter, ex SCID mice were trnsferred with 4x1 nïve CD4 + CD4RB hi T cells isolted from spleens of wild-type SPF BALB/c mice., Supplementtion with 17-mix significntly suppressed wsting disese-ssocited weight loss. Grph shows weight curves of SCID recipients. Ech curve represents the verge weight of mice +/- s.e.m., The numers of CD4 + + cells in the colon LP of SCID nd SCID mice on dy 14 fter T cell trnsfer re shown. Trnsfer of nïve CD4 T cells into SCID recipients induces intestinl inflmmtion, nd this is ccompnied y the induction of smll popultion of + Treg cells. Although not sttisticlly significnt, there ws tendency towrds n increse in + Treg cells numers in SCID mice. c, The distl intestines were collected from mice in ech group on dy 14 fter the T cell trnsfer nd exmined for expression of, T-β1, T-β, IL-6 nd TNFα mrna y quntittive PCR. There ws significntly incresed expression of T-β mrna in SPF mice compred with control mice, s well s tendency towrds n increse of nd T-β1 nd reduction of inflmmtory cytokine trnscripts. Dt re representtive of two independent experiments. Error rs, s.d. P <.1; P <., s clculted y the Student s t-test. 17
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doi:10.1038/nture09663 Scrmle shnlrp3 shcsp1 IL-1β (p17) IL-1β (pg/ml) 2000 1500 1000 500 Wt Nlrp3-/- Ipf-/- 0 APDC IL-1β (p17) Supplementl Figure 1. Mitochondril ROS cn trigger NLRP3 inflmmsome ctivtion,