RESEARCH ARTICLES ◥
CELIAC DISEASE
Reovirus infection triggers
inflammatory responses to dietary
antigens and development of
celiac disease
Romain Bouziat,1,2 Reinhard Hinterleitner,1,2 Judy J. Brown, 3, 4*
Jennifer E. Stencel-Baerenwald, 3, 4 Mine Ikizler, 4, 5 Toufic Mayassi,1,2 Marlies Meisel,1,2
Sangman M. Kim,1,2 Valentina Discepolo,1, 6 Andrea J. Pruijssers, 4, 5 Jordan D. Ernest,1,2
Jason A. Iskarpatyoti, 4, 5 Léa M. M. Costes,1, 7 Ian Lawrence,1,2 Brad A. Palanski, 8
Mukund Varma, 9, 10 Matthew A. Zurenski,1,2 Solomiia Khomandiak, 4, 5
Nicole McAllister, 3, 4 Pavithra Aravamudhan, 4, 5 Karl W. Boehme, 4, 5 Fengling Hu,1
Janneke N. Samsom, 7 Hans-Christian Reinecker, 9 Sonia S. Kupfer,1,11
Stefano Guandalini,11, 12 Carol E. Semrad,1,11 Valérie Abadie, 13 Chaitan Khosla, 8, 14, 15
Luis B. Barreiro, 16 Ramnik J. Xavier, 9, 10, 17 Aylwin Ng, 9, 10
Terence S. Dermody, 3, 4, 5, 18, 19† Bana Jabri1,2,11, 12, 20†
Viral infections have been proposed to elicit pathological processes leading to the
initiation of T helper 1 (TH1) immunity against dietary gluten and celiac disease (CeD). To
test this hypothesis and gain insights into mechanisms underlying virus-induced loss of
tolerance to dietary antigens, we developed a viral infection model that makes use of
two reovirus strains that infect the intestine but differ in their immunopathological
outcomes. Reovirus is an avirulent pathogen that elicits protective immunity, but we
discovered that it can nonetheless disrupt intestinal immune homeostasis at inductive and
effector sites of oral tolerance by suppressing peripheral regulatory T cell (pTreg)
conversion and promoting TH1 immunity to dietary antigen. Initiation of TH1 immunity to
dietary antigen was dependent on interferon regulatory factor 1 and dissociated from
suppression of p Treg conversion, which was mediated by type-1 interferon. Last, our study
in humans supports a role for infection with reovirus, a seemingly innocuous virus, in
triggering the development of CeD.
Celiac disease (CeD) is a complex immune disorder with an autoimmune component in which genetically susceptible individuals expressing the human leukocyte antigen (HLA) DQ2 or DQ8 molecules display an in-
flammatory T helper 1 (TH1) immune response
against dietary gluten present in wheat (1– 3). The
HLA-DQ2– or HLA-DQ8–restricted TH1 response
against gluten is central to CeD pathogenesis and
thought to precede development of villous atro-
phy ( 4). However, epidemiological and immuno-
logical observations support a role for additional
genetic and environmental factors in CeD patho-
genesis. Similar levels of wheat consumption and
expression of CeD-predisposing HLA molecules
can be accompanied by striking differences in CeD
prevalence (1). A remarkable example supporting
a role for environmental factors is the high frequen-
cy of CeD in Finnish Karelia (>2%), which con-
trasts with the low incidence of CeD in the adjacent
Russian republic of Karelia (0.2%), two neighbor-
ing regions harboring genetically similar popula-
tions. Furthermore, the incomplete digestion of
gluten by intestinal enzymes (2, 5) explains why
gluten would be conducive to inducing intestinal
T cell responses. However, it does not explain why
CeD patients develop a gluten-specific TH1 response
instead of a regulatory immune response, the de-
fault intestinal immune reaction to orally ingested
protein, characterized by the induction of periph-
eral regulatory T cells (p Tregs) expressing the tran-
scription factor forkhead box P3 (Foxp3) ( 6).
Viral infection experimental model using
genetically engineered reoviruses
Despite epidemiological evidence of associations
between viral infections and the initiation of CeD
(1), experimental evidence is lacking. Previous studies
have implicated adenovirus, enteroviruses, hepatitis C virus, and rotavirus as triggers of CeD ( 7, 8).
However, little is known about the mechanisms
by which viruses evoke the disease. Viruses in the
family Reoviridae are segmented, double-stranded
RNA (dsRNA) viruses that infect humans frequently throughout their lifetime ( 9). Mammalian
Orthoreovirus (reovirus) strains isolated from humans
can infect mice via the oral route and activate innate immune pathways similar to the related rotavirus ( 10, 11). Two human reovirus isolates, type
1 Lang (T1L) and type 3 Dearing (T3D), differ in
replication biology, apoptosis induction, innate immune response activation, cellular tropism, and
pathogenesis (11). Furthermore, T1L infects the
intestine and perturbs intestinal immune homeostasis (11, 12), whereas T3D is incapable of infecting the intestine (11). On the basis of the fundamental
differences between T1L and T3D, we hypothesized that engineering a T3D reassortant virus
capable of intestinal infection would yield two viruses with potentially different effects on tolerance to dietary antigen. Therefore, we recovered a
T3D reassortant virus called T3D-RV by introducing the S1 and L2 gene segments of T1L into a
T3D genetic background, thus allowing the virus
to infect the intestine (fig. S1A) ( 13). Such reassortants arise naturally ( 10, 11) and can be readily
recovered in the laboratory by using reverse genetics (11). We first established that the two viruses are similar in their capacity to replicate (fig.
S1B) and infect the intestine (fig. S1, C and D). Furthermore, both viruses are cleared (fig. S1E) without
inducing intestinal damage (fig. S1F). Although
both viruses induced high antireovirus antibody
titers, antibody levels observed after T1L infection
were significantly higher than those after T3D-RV
infection (fig. S1G). However, comparison of the
host T cell response in sham- and virus-infected
mice revealed that T1L and T3D-RV induced similar TH1 responses in Peyer’s patches (PP) (fig. S1H),
RESEARCH
44 7 APRIL 2017 • VOL 356 ISSUE 6333 sciencemag.org SCIENCE
1Department of Medicine, University of Chicago, Chicago, IL, USA. 2Committee on Immunology, University of Chicago, Chicago, IL, USA. 3Department of Pathology, Microbiology, and Immunology,
Vanderbilt University Medical Center, Nashville, TN, USA. 4Elizabeth B. Lamb Center for Pediatric Research, Vanderbilt University Medical Center, Nashville, TN, USA. 5Department of Pediatrics,
Vanderbilt University Medical Center, Nashville, TN, USA. 6Department of Translational Medical Sciences, Section of Pediatrics, University of Naples Federico II, and CeInGe–Biotecnologie
Avanzate, Naples, Italy. 7Laboratory of Pediatrics, Division of Gastroenterology and Nutrition, Erasmus University Medical Center Rotterdam-Sophia Children’s Hospital, Rotterdam, Netherlands.
8Department of Chemistry, Stanford University, Stanford, CA, USA. 9Division of Gastroenterology, Department of Medicine, Gastrointestinal Unit and Center for the Study of Inflammatory Bowel
Disease, Massachusetts General Hospital and Harvard Medical School, Boston, MA, USA. 10Broad Institute of MIT and Harvard University, Cambridge, MA, USA. 11University of Chicago Celiac
Disease Center, University of Chicago, Chicago, IL, USA. 12Section of Gastroenterology, Hepatology, and Nutrition, Department of Pediatrics, University of Chicago, Chicago, IL, USA. 13Department
of Microbiology, Infectiology, and Immunology, University of Montreal, and the Centre Hospitalier Universitaire (CHU) Sainte-Justine Research Center, Montreal, Quebec, Canada. 14Department of
Chemical Engineering, Stanford University, Stanford, CA, USA. 15Stanford ChEM-H, Stanford University, Stanford, California, USA. 16Department of Genetics, CHU Sainte-Justine Research Center,
Montreal, Quebec, Canada. 17Center for Computational and Integrative Biology, Massachusetts General Hospital and Harvard Medical School, Boston, MA, USA. 18Department of Pediatrics,
University of Pittsburgh School of Medicine, Pittsburgh, PA, USA. 19Department of Microbiology and Molecular Genetics, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA.
20Department of Pathology, University of Chicago, Chicago, IL, USA.
*These authors contributed equally to this work. †Corresponding author. Email: bjabri@bsd.uchicago.edu (B.J.); terence.dermody@chp.edu (T.S.D.)
