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Scandinavian Journal of Gastroenterology, 2010; 45: 160–167
ORIGINAL ARTICLE
Helicobacter species DNA in liver and gastric tissues in children and
adolescents with chronic liver disease
THOMAS H. CASSWALL
1
, ANTAL NÉMETH
1
, INGRID NILSSON
2
,
TORKEL WADSTRÖM
2
& HANS-OLOF NILSSON
2
1
Department of Pediatric Gastroenterology, Hepatology and Nutrition, Astrid Lindgren’s Children’s Hospital at
Karolinska University Hospital, CLINTEC, Karolinska Institutet, Stockholm, Sweden, and
2
Department of Medical
Microbiology, University of Lund, Lund, Sweden
Abstract
Objective. Enterohepatic Helicobacter species (EHS) have previously been found in adults with hepatobiliary diseases. Here,
we report the prevalence of Helicobacter pylori and EHS in liver and gastric tissue in children and adolescents with chronic liver
disease (CLD). Material and methods. Seventy-seven consecutive children and adolescents with CLD with or without
ulcerative colitis or Crohn’s disease (UC/CD) were investigated. Tissue samples were analysed using a Helicobacter genus-
specific 16S rDNA polymerase chain reaction (PCR) assay and DNA-sequence analysis. Sera from 61 subjects were also
analysed using enzyme immunoassay and immunoblotting. Results. The Helicobacter PCR was positive in 3/23 (13%) livers
from patients with primary sclerosing cholangitis and UC, and in 1/2 livers from patients with autoimmune hepatitis (AIH)
and UC. Sequenced PCR products matched the 16S rDNA of H. hepaticus,H. muridarum,H. canis, and H. pylori, respectively.
H. ganmani and H. bilis were detected in gastric tissues from two AIH patients. H. hepaticus and H. pullorum were found in livers
from two patients with acute liver failure and intrahepatic cholestasis. Antibody reactivity to Helicobacter cell-surface proteins
was negative. Conclusions. H. pylori and EHS can be detected in the livers of some patients with UC and concomitant liver
disease, as well as in other children with liver diseases. Multicentre studies from different locations are needed to find out
whether these bacteria play a pathogenetic role or whether their presence is an epiphenomenon.
Key Words: Autoimmune hepatitis, children, enterohepatic Helicobacter species, Helicobacter pylori, inflammatory bowel
disease, liver tissue, polymerase chain reaction, primary sclerosing cholangitis, ulcerative colitis
Introduction
Since the discovery of Helicobacter pylori by Warren
and Marshall 27 years ago, at least 28 different
Helicobacter species have been characterized. It seems
that most mammals, but also many other animals and
birds, are colonized with various gastric and enteric
Helicobacter species in the gut [1]. During the last
decade, many non-H. pylori Helicobacters have been
isolated from samples of the human stomach and
intestine, as well as from blood and faeces [2]. These
so-called enterohepatic Helicobacter species (EHS)
may cause chronic inflammation in the distal bowel
and hepatobiliary tract in rodents, dogs, primates and
birds [3]. A number of recent studies also found an
association between EHS and human hepatobiliary
tract disease [2]. Since most Helicobacter species are
difficult to culture, polymerase chain reaction (PCR),
serology and immunohistochemistry have been used
for diagnosis [4–7], but successful culture from dis-
eased livers has also been reported [8]. Patients suf-
fering from hepatobiliary diseases such as
hepatocellular carcinoma, primary sclerosing cholan-
gitis (PSC) and primary biliary cirrhosis (PBC) [6,9],
as well as chronic biliary tract disease and cancer, have
been investigated for EHS [10,11].
In the industrialized world, PSC and autoimmune
hepatitis (AIH) belong to the more common causes of
Correspondence: Thomas H. Casswall, MD, PhD, Division of Pediatric Gastroent erology, Hepatology and Nutrition, Astrid Lindgren’s Children’s Hospital B57,
Karolinska University Hospital, SE-141 86 Stockholm, Sweden. Tel: +468 585 81464. Fax: +468 585 814 10. E-mail: thomas.casswall@ki.se
(Received 17 July 2009; accepted 19 October 2009)
ISSN 0036-5521 print/ISSN 1502-7708 online 2010 Informa UK Ltd.
DOI: 10.3109/00365520903426915
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paediatric liver disease, mostly occurring during the
second decade of life. Such progressive liver conditions
may lead to end-state liver disease (ESLD). Their basic
aetiology is largely unknown. Recently, it has been
suggested that the primary triggering factor might be
auto- as well as allo-immunogenic, thereby dividing the
disease into autoimmune and immune-modulated
inflammatory disorders, respectively [12]. In the latter
case, bacterial or viral agents could serve as triggering
factors. Thus, in the intestinal mucosa of patients with
inflammatory bowel disease (IBD), cytomegalovirus
has been found to an extent that makesthe pathogenetic
role of this virus highly probable [13]. The pathogenesis
also varies and PSC especially can have both autoim-
mune(AI)-likeandnon-AI forms. Thenaturalcourseof
juvenile AIH without treatment is generally poor [14],
while the outcome of juvenile PSC varies and is partly
unknown. However, data so far show that a high pro-
portionofpatientswithjuvenile PSC willdevelopESLD
[15]. More than 80% of children with PSC also suffer
from IBD, mostly ulcerative colitis (UC) [16].
Whether EHS are involved in the pathogenesis of
chronic liver diseases (CLDs) in children and ado-
lescents is unknown. However, we have previously
identified EHS (H. ganmani) from livers of children
with various CLDs [17]. The previous study was
performed on archived paraffin-embedded tissue
samples using PCR and DNA sequencing. In the
present study, we investigated fresh liver and stomach
samples, as well as serum samples, from children and
adolescents undergoing routine evaluation for CLDs,
with special emphasis on PSC and AIH.
Material and methods
Patients
Seventy-seven consecutive children and adolescents
(34 males) below the age of 19 years were included,
with either AIH (Group 1; n=24; 10 males; mean age
15.6 years), PSC (Group 2; n=27; 14 males; mean age
14.4 years) or IBD and liver damage (Group 3; 1 male;
14 years old). The diagnosis of AIH was based on the
following criteria: clinical signs, biochemical evidence
of non-cholestatic liver damage, evidence of autoanti-
bodies, elevated IgG and histological features accord-
ing to the International Autoimmune Hepatitis Score
(IAIHS) [18]. The diagnosis of PSC was establis hed by
means of liver function profile, endoscopic retrograde
cholangiopancreatography/magnetic resonance cho-
langiopancreatography (MRCP) and liver biopsy.
The diagnosis of IBD was based on macro- and
microscopic findings obtained by colonoscopy and
upper endoscopy. Colonoscopy was only performed
in patients with symptoms and signs indicating IBD.
Three of the 27 patients with PSC had no evidence of
IBD. Although 19 of these 27 patients had either
antinuclear and/or anti-smooth muscle antibodies,
only two of them fulfilled the criteria for “possible
autoimmune disease”according to the IAIHS.
Patients in Groups 1 and 2 were treated according to
the established guidelines, depending on the diagnosis,
and treatment included immunosuppression and urso-
deoxycholicacid if needed. Furthermore,children with
various liver diseases were included as disease cont rols:
Group 4 (n=25; nine males; mean age 8.6 years).
Progressive familial intrahepatic cholestasis (PFIC)
was the most common individual disease in these chil-
dren (n=10). The various diagnoses and demographic
data for all the patients are presented in Table I. Liver
biopsies were collected from all 77 patients, whereas
tissuesamples of the gastric mucosa were available in 23
of them (29.9%). Serum samples from 61 patients
(79%) were available for analyses of antibodies against
H. pylori and three EHS by means of enzyme immu-
noassay (EIA) and immunoblotting (IB). In Groups 1
and 2, 18 children (n=7 and n=11, respectively) were
immunosuppressive treatment naive patients, one
Group 1 patient received steroids 6 weeks before diag-
nosis and in 33 patients the samples were taken as
protocol biopsies during routine follow-up after a
medianof2.8 years(range0.5–10 years)after diagnosis.
All samples were collected during 2002–06. Informed
consent was obtained from the patient, parent or both.
Thestudy wasapproved bythe RegionalEthical Review
Board in Stockholm (No. 48-02).
Tissue biopsies and serum samples
Transcutaneous liver biopsies were obtained using a
17 G (1.2 mm) or 18 G (1.4 mm) Hepafix
liver
biopsy needle (Braun Melsungen AG, Germany).
A1–2-mm piece of the liver sample was wrapped
in aluminium foil, put into a small glass tube with a
screw-cap lid, snap-frozen in liquid nitrogen and
stored at –80C. From the patients who underwent
routine upper endoscopy, one biopsy was taken from
both the antrum and corpus areas of the gastric
mucosa, aseptically wrapped in aluminium foil and
treated the same way as the liver biopsies. For the
serological analyses, when applicable, 2 ml of the
patient serum was stored at –20C. All the samples
were later transported on dry ice to Lund University,
where they were kept at –20C until analysed.
Helicobacter genus-specific PCR and DNA sequencing
DNA was extracted from liver and gastric tissue
samples using the QIAamp DNA Mini-Kit (Qiagen,
Helicobacter species DNA in liver tissues 161
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Hilden, Germany) according to the manufacturer’s
instructions. Approximately 5 mg of each tissue sam-
ple was used. DNA extracts were amplified in a
GeneAmp 2700 Thermocycler (Applied Biosystems,
Foster City, CA) using a semi-nested PCR
assay specific for Helicobacter species 16S rDNA, as
previously described [17], using primers 1F (5-
CTATGACGGGTATCCGGC-3), 1R (5-CTCAC-
GACACGAGCTGAC-3) and 2R (5-TCGCCT-
TCGCAATGAGTATT-3), as constructed by Goto
et al. [19]. Primers 1F and 1R were used in the first
step, whereas primers 1F and 2R were used in the
second step. PCR products were visualized as previ-
ously described [15]. DNA sequencing, sequence
alignment and determination of closest known partial
16S rDNA relatives were performed as previously
described [17,20].
Serum H. pylori EIA
Sera were tested for IgG antibodies against H. pylori
cell-surface proteins (CSPs) using a standardized EIA
[21]. The results were recorded as relative antibody
activities (RAAs). Corrected mean absorbance values
as percentages of the absorbance measured with a
human gamma globulin (Pharmacia & UpJohn,
Stockholm, Sweden) were used as reference stan-
dards. Samples giving RAAs of >35 and <25 were
considered seropositive and seronegative, respec-
tively. The cut-off values were based on the results
of EIA and IB analyses in which sera from Swedish
subjects (H. pylori culture-positive or -negative
patients, as well as healthy blood donors and children)
were tested.
Serum enteric Helicobacter EIA
Sera were tested for IgG antibodies to CSPs of
H. pullorum (strain CCUG 33838), H. bilis (strain
CCUG 38995) and H. hepaticus (strain
CCUG33637) using EIA methods, as previously
described [22]. To minimize cross-reactivity between
H. pylori and the other Helicobacter species, serum
samples were absorbed prior to testing. For the
enteric Helicobacter species, cut-off values (corre-
sponding to the upper limit for the 95th percentile
Table I. Diagnostic groups, demographic data of the patients and the results of Helicobacter genus-specific 16S rDNA PCR from liver and
gastric biopsies.
Diagnosis
No. of
patients
No. of
boys
Age (years);
median (range)
No. of
PCR-positive
liver biopsies
No. of
PCR-positive
gastric biopsies
Serum available
from total
no. of patients
Naïve
patients
f
Group 1 (AIH) 24 10/24 15.6 (2.2–18.6) 1/24 3/8 23/24 7/23
No IBD 22
IBD 2
a
1/2
Group 2 (PSC) 27 14/27 14 (8–18.9) 3/27 1/8 20/27 6/18
No IBD 3
IBD 23
b
3/23
Coeliac disease 1
Group 3 (IBD + liver damage) 1
c
1/1 14 1/1
Group 4 (disease control patients) 25 9/25 8.5 (1.4–17) 2/25 1/7 17/25
Intrahepatic cholestasis 10
d
1/10
AATD 3
Recurrent episodes of transitory
extrahepatic cholestasis
3
Other 9
e
1/9
Total, all patients (range) 77 34/77 13.5 (1.4–18.9) 6/77 5/23 61/77
a
UC.
b
UC, n=18; Crohn’s disease, n=4; indeterminate colitis, n=1.
c
UC.
d
Progressive familial intrahepatic cholestasis, n=8; Alagille’s syndrome, n=1; Aagenaes syndrome, n=1.
e
One each of biliary atresia, celiac disease, liver steatosis, non-specific mitochondriopathy, acute liver failure, OTC deficiency, portal vein
thrombosis, toxic epidermal necrolysis with liver damage and mild non-progressive liver disease.
f
Patients who had not received immunosuppressive treatment before first investigation.
AATD =a-1-antitrypsin deficiency.
162 T. H. Casswall et al.
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level) were established based on EIA results obtained
using sera from 90 healthy blood donors. Cut-off
values were ‡25 for H. pullorum and H. hepaticus
and ‡35 for H. bilis.
Serum IB assays
Sera giving positive or borderline results in the EIAs
were further analysed by IB for diagnosis of H. pylori
[23], as well as for H. pullorum,H. bilis and
H. hepaticus [24]. The same Helicobacter extracts used
as antigens in the EIAs were used in the IB analyses.
In the subsequent IB, the intensities of the bands
obtained with each test serum were compared with
those obtained using a reference strip incubated with a
polyclonal antiserum against each of the four Helico-
bacter species. Criteria for Helicobacter-positive immu-
noblots were followed as previously described [23].
In total, serum was available from 59/77 patients
(79%) [AIH with or without IBD, n=23; PSC with or
without IBD, n=18; IBD+non-specific liver disease,
n=1; PFIC, n=10; other liver diseases, n=7
(Table I)]. Sera were screened using EIA and fol-
lowed up with IB when borderline or positive EIA
results were obtained.
Results
The results of the Helicobacter genus-specific 16S
rDNA from the diagnostic groups are presented
in Table I. The Helicobacter genus-specific PCR assay
was positive in liver biopsies from 6/77 patients
(7.8%): 1/24 (4.2%) in Group 1, 3/27 (11.1%) in
Group 2 and 2/25 (8%) in Group 4. The Helicobacter
PCR-positive patients with PSC and AIH all
presented with concomitant UC. Four of 20 patients
(20%) with PSC or AIH and concomitant UC were
Helicobacter-positive in the liver, compared with 2/57
liver disease patients without UC (3.5%) (p=0.06;
Fischer’s exact test). DNA sequencing and Basic
Local Alignment Search Tool nucleotide (BLASTn)
analysis of PCR products from liver samples of
patients with PSC–UC (n=3) and AIH–UC
(n=1) revealed H. hepaticus,H. muridarum,H. pylori
and H. canis (Table II).
Gastric biopsies were available in 23 patients, of
which five (22%) were PCR-positive for Helicobacter
species. Four of these patients were from Groups 1 and
2. DNA sequencing of gastric Helicobacter genus-
specific PCR products in two AIH patients with a Heli-
cobacter-negative liver sample demonstrated H. pylori
and H. ganmani, respectively. A sequence similarity
search identified H. bilis inthe gastric mucosaof apatient
with AIH and concomitant UC. One patient with PSC
and concomitant UC was positive in both liver tissue
(H.muridarum)andgastricmucosa(H.pylori)(TableII).
The results from the disease control group are also
presented in Tables I and II. All Helicobacter species
found in different disease categories and the type of
specimen tested are presented in Table III.
Twelve of the 61 available sera showed increased or
borderline increased IgG antibody levels by EIA
and four of these (6.6%) were IB-positive for H. pylori.
The IB-positive sera were from two AIH patients
with a PCR-negative liver sample and for whom
no gastric biopsies were available. One patient with
ornithine transcarbamylase (OTC) deficiency was
PCR-negative in the liver but H. pylori-positive in
the stomach and IB-positive for H. pylori. Another
patient with AIH–UC and a PCR-positive liver sam-
ple (H. canis), but a negative gastric biopsy, was H.
pylori IB-positive.
Table II. Helicobacter DNA sequence analysis of positive PCR-products from liver and gastric tissues as well as results of IB analysis.
Patient No. Diagnosis Age (years) Sequence similarity: liver Sequence similarity: stomach IB EIA
1 AIH 12.6 Negative H. pylori Negative Negative
2 AIH 15 Negative H. ganmani Negative Negative
3 AIH+UC 15.6 Negative H. bilis Negative Negative
4 AIH+UC 16 H. canis Negative Positive Negative
5 PSC+UC 15 H. pylori Negative Negative Negative
6 PSC+UC 16.8 H. muridarum H. pylori nd Negative
7 PSC+UC 18.8 H. hepaticus nd nd Negative
Disease controls
8 Acute liver failure 2.8 H. hepaticus Negative Negative Negative
9 OTC deficiency 12.6 Negative H. pylori Positive Negative
10 PFIC 1.4 H. pullorum nd Negative Negative
nd =not done.
Helicobacter species DNA in liver tissues 163
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None of the sera showed any antibody reactivity to
antigens of the three EHS analysed in this study.
H. pylori infection could not be confirmed by his-
tology, rapid urease test or culture in any of the three
children who were PCR-positive for H. pylori in
gastric samples. Serum from one of these patients
was positive for H. pylori by EIA and IB.
Discussion
There is growing awareness of the role of enteric
Helicobacter in human liver disease. Several studies
of adult patients have analysed the prevalence of
Helicobacter species in different CLDs. H. pylori has
been detected in liver tissues from humans with CLDs
such as chronic hepatitis C virus [5], liver cancer [25]
and gallbladder disease [26]. Although H. pylori may
be able to survive in low concentrations of human bile
[27], other Helicobacter species, such as H. hepaticus,
H. bilis and H. pullorum, are much more bile-tolerant
[28], which may in part explain why these EHS have
been detected in human livers [9,29].
In a previous study, we have detected Helicobacter
species with PCR and DNA sequencing in livers from
adults suffering from PSC and PBC [9]. However,
only H. pylori and no EHS could be found.
There are limited data available on EHS in paedi-
atric patients. We previously showed, by species-
specific PCR analysis and DNA sequencing, that
40/61 samples (65.6%) from children with various
liver diseases were positive for H. pylori and H. gan-
mani [17]. These results were obtained in children
from Detroit, MI and only 33% of them were Cau-
casian, as compared to the present study in which 72/
77 were native Swedes. The prevalence of H. pylori
in native Swedish-born schoolchildren is only 2%
[30]. The findings in the study of Tolia et al. [17]
might thus have been influenced by geographical
localization, patient ethnicity [30] or disease pattern.
In an Australian study, EHS have been detected in
children with IBD and irritable bowel syndrome by
fluorescent rRNA in situ hybridization [31]. An
altered gut flora in an inflamed intestine, as in IBD
and especially UC, may predispose to gut transloca-
tion and transmission of H. pylori and EHS to the
liver.
In this study we detected DNA of Helicobacter
species in livers from six children and EHS from
the stomach mucosa from another two children. In
the disease groups (AIH, PSC and IBD+liver dam-
age), all positive livers were from patients with AIH or
PSC in combination with UC. On the other hand,
23/24 livers from children with AIH were negative; a
liver biopsy from a child with AIH and concomitant
UC was positive for H. canis. EHS were found in 5/25
livers (25%) from IBD patients. Consequently, there
was a trend (p=0.06) that coexistence of UC with
liver disease appeared to increase the probability of
finding Helicobacter in liver biopsies. EHS DNA was
also found in one child with acute liver failure (H.
hepaticus) and one with PFIC (H. pullorum) (Table II).
Sixty-one of the paediatric and adolescent patients
were screened for IgG serum antibody levels to H.
pylori and three EHS with two different immunoas-
says (EIA and IB). Increased H. pylori antibody levels
were measured in 6.6% of tested sera. The seroprev-
alence in asymptomatic native Swedish children varies
between 4% and 10% depending on age [32]. Two of
these 61 patients were found positive by PCR for EHS
in liver tissue; however, no seroconversion was
detected. H. canis was detected by PCR in the liver
of one patient with AIH and UC but showed antibody
reactivity to H. pylori, which might be due to cross-
reactivity between similar antigens of these two gastric
species.
We detected Helicobacter species in livers from 3/27
patients with PSC (11.1%), but only if associated
with UC. Thus, Helicobacter species, along with pos-
sibly other microbes, may only partly be involved in
the pathogenesis of PSC.
PSC is a progressive obliterative inflammation and
fibrosis of the intra- and extrahepatic bile ducts, which
may progress to liver fibrosis and cirrhosis. The out-
come of childhood disease is less well known, but
seems to be as severe as for adults [15]. In the
Table III. All Helicobacter species found in serum, liver and stomach according to diagnostic group. Values shown represent numbers of
patients.
Disease category Positive/total Serum only Serum and liver Stomach only Stomach and liver Liver only
AIH 4/24 0/24 1/24 3/24 0/24 0/24
PSC (all categories) 3/27 0/27 0/27 0/27 1/27 2/27
IBD (all categories)
a
5/25 0/25 0/25 1/25 1/25 4/25
Disease controls 3/25 0/25 0/25 1/25
b
0/25 2/25
a
IBD+AIH or PSC.
b
Stomach+serum.
164 T. H. Casswall et al.
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paediatric population, approximately 80% of patients
have concomitant IBD of the colon, where UC is by
far the most common type (»60%) [15]. The aetiol-
ogy of PSC is unknown, but one suggested pathoge-
netic mechanism might be the translocation of
bacterial antigens through the intestinal epithelium
of the inflamed colon into the biliary tree via the portal
circulation. Finally, these antigens are presented in
the liver to T lymphocytes, which activate auto-
reactive T or B cells [33]. Evidence of translocation
into bile is supported by positive bacterial cultures
from bile of PSC patients as compared to PBC
patients [34]. The isolation of H. cinaedi from the
colon, liver and mesenteric lymph nodes of a
Rhesus monkey further supports the translocation
hypothesis [35], and H. pylori erythrocyte binding
has been suggested as a plausible mechanism for
disseminated disease [36]. H. pylori has recently
been shown in vitro to adhere to and invade human
hepatocytes and to persist within the liver cells during
subcultures. This internalization was also shown
to be more efficient in hepatocytes than in gastric
epithelial cells [37].
The pathogenesis of AIH is different from that of
PSC [38], which may explain why Helicobacters were
not found in these patients except when UC was
present. Whether Helicobacter species only reflect a
disturbed gut flora, or if they are to some extent
involved in the pathogenic process, has to be further
evaluated.
It is unclear why EHS could be detected in human
livers. The bacteria naturally colonize domestic ani-
mals, as well as laboratory rodents such as mice,
gerbils and rats. A zoonotic potential for some
Helicobacter species has been proposed [39]. The
natural animal hosts of these EHS, together with
the human diseases in which they have been detected,
are summarized in Table IV.
For a paediatric cohort, our sample is rather large.
However, EHS were found in livers from only six
children, and thus caution should be used when
interpreting the findings. Another limitation of the
study is the lack of healthy children as controls. It is
obvious that this is ethically unacceptable unless post-
mortem samples are used. Thus, the prevalence of
Helicobacter in the livers of aged-matched healthy and
diseased control subjects is presently unknown. Stud-
ies published so far are conflicting, and only one
Helicobacter strain has been cultured from a liver
with Wilson’s disease [40].
The exact role of EHS and H. pylori in the path-
ogenesis of chronic inflammatory liver and bowel
disease remains unclear. Due to the small numbers
of EHS found, caution has to be exercised regarding
their relevance in the pathogenesis of liver and gut
diseases. Worldwide epidemiological data for H.
pylori also indicate large variations in the figures
for EHS.
In order to clarify the pathogenetic importance of
EHS, future studies should be conducted in a multi-
centre manner including a larger number of patients
(with AIH and PSC, both with and without IBD)
from different parts of the world.
In conclusion, this study demonstrated that Heli-
cobacter spp DNA can be detected in liver and
stomach tissue specimens using PCR. Using our
data, we could not decide whether this is only an
epiphenomenon or whether the EHS play a patho-
genetic role. Only considerably larger multicentre
studies from different geographical locations can
do this.
Acknowledgement
This study was supported by the Karolinska Univer-
sity Hospital, Stockholm, Lund University Hospital
Table IV. Helicobacter species other than H. pylori found in livers or gastric samples from patients with liver disease with or without IBD, their
proven animal hosts and their potential role in human disease. Findings from the present study and elsewhere.
Helicobacter species Animal host and localization [45,46] Human disease
H. bilis Mice: intestine and liver; gerbils: intestine AIH+UC (patient No. 3), chronic cholangitis,
gallbladder cancer [10,41]
H. canis Dogs and cats: intestine and liver [42,43] AIH+UC (patient No. 4), diarrhoeal disease [44]
H. ganmani Mice: intestine AIH (patient No. 2), BA, HCV, PSC, steatohepatitis [17]
H. hepaticus Mice: intestine and liver PSC+UC (patient No. 7), chronic cholangitis, AIH [41]
H. muridarum Rats: intestine PSC+UC (patient No. 6)
H. pullorum Poultry: broiler chicken carcasses and intestines
from laying hens
PFIC (patient No. 10), gastroenteritis and healthy subjects
BA =biliary atresia; HCV =hepatitis C virus.
Helicobacter species DNA in liver tissues 165
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(ALF grant), The Royal Physiographic Society in
Lund and the Swedish order of Freemasons.
Declaration of interest: The authors declare no
conflict of interest.
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