Science - USA (2021-11-05)

Huet al.,Science 374 , eabe6723 (2021) 5 November 2021 6 of 13

B

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Candidatus Arthromitus

Lactobacillus

Turicibacter

Clostridiales-2

Coriobacteriaceae

Allobaculum

S24-7

Sutterella

Desulfovibrio

Ruminococcus

Akkermansia

TM7

Bifidobacterium

WT

Ileum lumen

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WT

Colon lumen

-2 -1 0 1 2

D

Lumen Mucus-associated

C

16 SrDNA DAPI

(universal probe)

WT

ileum

colon

** *** ** ** * ***

ns ns ns ns ns ns

WT WT WT WT WT WT

Lactobacillus SFB Bacteroidetes Lactobacillus SFB Bacteroidetes

103

104

105

106

102

103

104

105

106

102

1.0

1.5

2.0

2.5

0.5

101

102

103

104

100

105

103

104

105

106

102

103

104

105

106

102

102

103

104

105

101

102

103

104

105

101

102

103

104

105

101

(x10

5 )

Taxon-specific 16

S

rDNA

(relative units)

A

Bacilli

Clostridia

Erysipelotrichi

Actinobacteria

Coriobacteriia

Bacteroidia

Verrucomicrobiae

Betaproteobacteria

Deltaproteobacteria

Deferribacteres

Others

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0

20

40

60

80

100

Relati

ve abundance (%)

Ileum lumen Ileum mucus Colon lumen Colon mucus

WT WT WT WT z-score

Small intestine

Sprr2a/ Sprr2a/ Sprr2a/ Sprr2a/

Sprr2a/ Sprr2a/

Sprr2

a

/

Sprr2a

/

Sprr

2a

/

Sprr2a

/

Sprr2

a

/

Sprr

2a

/

Sprr2a

/

103

104

105

106

102

103

104

105

106

102

101

102

103

104

100

105

EF

Mesenteric

lymph nodes

CFU/g tissue

Liver Spleen

** ** p=0.085

105

100

101

102

103

104

106

0246810

0

20

40

60

80

100

days

% Surv

iv

al

*

WT

(n=9)

Sprr2a/

(n=8)

L. monocytogenes infection

107

WT

Sprr2

a

/

WT

Sprr2

a

/

WT

Sprr2a

/

Fig. 3. Mice lacking SPRR2A have an altered intestinal microbiota and are
more susceptible toL.monocytogenesinfection.(A) Male WT andSprr2a−/−
littermates were separated at weaning and caged by genotype for 6 weeks.
Intestinal mucus–associated and luminal microbial communities were charac-
terized by 16SrRNA sequencing. Relative abundances of bacterial classes in WT
andSprr2a−/−mice are shown.n= 4 mice per group. (B) Relative abundances of
bacterial genera in WT andSprr2a−/−mice. (C) qPCR analysis of specific
bacterial groups in the intestinal mucus–associated and luminal microbial
communities of wild-type andSprr2a−/−littermates. Values for each bacterial
group are expressed relative to total 16SrDNA levels. P< 0.05; P< 0.01;
P< 0.001; ns, not significant by two-tailedttest. (D) Fluorescence in situ

hybridization (FISH) detection of bacteria in the small intestines of WT andSprr2a−/−

mice. Scale bars, 50mm. (E) 8- to 10-week-old WT (n= 12) andSprr2a−/−mice

(n= 14) were treated with antibiotics and orally infected with 1 × 10^9 CFU of

log-phaseL.monocytogenes. Liver, spleen, and mesenteric lymph nodes (MLN) were

collected after 24 hours, and bacterial counts were determined by dilution plating.

Results were pooled from three independent experiments. **P<0.01byMann-

WhitneyUtest. Dotted line indicates limit of detection. (F)8-to10-week-oldWT

(n= 9) andSprr2a−/−mice (n= 8) were orally infected with 2.5 × 10^9 CFU of log-

phaseL.monocytogenes. Survival rates were monitored over 10 days. Results were

pooled from two independent experiments. *P< 0.05 by the log-rank test. SFB,

segmented filamentous bacteria; CFU, colony-forming units.

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