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
12345678
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
12345678123456781234567812345678
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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