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Infection and Immunity, January 1999, p. 279-285, Vol. 67, No. 1
Third Department of Internal
Medicine1 and
Department of
Microbiology,
Received 28 May 1998/Returned for modification 29 July
1998/Accepted 22 October 1998
The immune responses to Helicobacter pylori infection
play important roles in gastroduodenal diseases. The contribution of gamma interferon (IFN- It is now generally accepted that
Helicobacter pylori infection is a major cause of chronic
gastritis and peptic ulcers. It has been suggested that the
histological damage due to the ammonia produced by H. pylori
urease (19), the vacuolization of gastric epithelium caused
by the cytotoxin derived from H. pylori (8), and
the mucosal injury caused by the host immune responses (1, 10, 15,
16) are important in the pathogenesis of these gastroduodenal diseases. Although it has been suspected that the activation of immune
cells is a pivotal factor and that cytokines contribute to their
activation, the mechanisms have not been clearly defined.
We previously investigated the cytokine expression patterns of human
gastric mucosal biopsy specimens by the reverse transcription-PCR method (20, 21). Since the results showed that the
expression of interleukin-8 (IL-8) mRNA was significantly higher in
H. pylori-positive gastritis than in H. pylori-negative controls and that there was a close correlation
between the expression of IL-8 mRNA and the severity of gastritis, we
suggested that IL-8 could play an important role(s) in mucosal inflammation.
In addition, gamma interferon (IFN- In an experiment with IFN- For investigations of the role of IFN- Animals.
IFN-
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Role of Gamma Interferon in Helicobacter
pylori-Induced Gastric Inflammatory Responses in a Mouse
Model
ABSTRACT
Top
Abstract
Introduction
Materials and methods
Results
Discussion
References
) to the immune responses, especially to the
induction of gastric inflammation and to protection from H. pylori infection, was investigated with IFN- gene knockout
(IFN-
/) mice. We first examined the colonizing
abilities of eight H. pylori strains with a short-term
infection test in order to select H. pylori strains which
could colonize the mouse stomach. Only three strains (ATCC 43504, CPY2052, and HPK127) colonized C57BL/6 wild-type mice, although all of
the strains except for ATCC 51110 could colonize
IFN-/ mice. The number of H. pylori
organisms colonizing the stomach in wild-type mice was lower than that
in IFN-/ mice. Oral immunization with the CPY2052
sonicate and cholera toxin protected against infection with strain
CPY2052 in both types of mouse. These findings suggested that IFN-
may play a protective role in H. pylori infection, although
the degree of its protective ability was estimated to be low. In
contrast, in a long-term infection test done to examine the
contribution of IFN- to gastric inflammation, CPY2052-infected
wild-type mice developed a severe infiltration of mononuclear cells in
the lamina propria and erosions in the gastric epithelium 15 months
after infection, whereas CPY2052-infected IFN-/ mice
showed no inflammatory symptoms. This result clearly demonstrated that
IFN- plays an important role in the induction of gastric inflammation caused by H. pylori infection.
INTRODUCTION
Top
Abstract
Introduction
Materials and methods
Results
Discussion
References
) and IL-1
mRNAs were detected
in the vast majority of the specimens examined in those studies,
whereas IL-2, IL-3, IL-4, IL-5, IL-9, IFN-, and tumor necrosis
factor alpha mRNAs were not detected in any specimens and IL-6 mRNA was
detected in only a few specimens. IFN- is thought to be important in
immune responses, because it induces the expression of the class II
major histocompatibility complex of antigen-presenting cells and
activates macrophages and natural killer cells.
gene knockout (IFN-/)
and wild-type mice, Dalton et al. showed that IFN- enhanced the
expression of the class II major histocompatibility complex on the
surface of peritoneal macrophages during Mycobacterium bovis
BCG infection and that IFN- contributed to protection from BCG
infection (2). Tagawa et al. reported that the development
of concanavalin A-induced hepatitis was reduced significantly in
IFN- gene knockout mice compared with that in wild-type mice
(18). Thus, IFN- appears to be involved in protection
from bacterial infection and in induction of inflammation. Although the
expression of IFN- mRNA was not specific to H. pylori-associated gastritis, the finding that the expression of
IFN- mRNA always took place in gastric mucosa (20) may
indicate that IFN- plays an important role(s) in the induction of
immune responses in gastroduodenal mucosa.
in H. pylori
infection, in vivo studies with IFN- gene knockout mice are an
effective means. Although colonization by H. pylori of the
mouse stomach had previously been considered difficult, Marchetti et
al. (12) and Lee et al. (11) recently identified
H. pylori strains with colonizing ability in the mouse
stomach. Marchetti et al. (12) showed, by examining the
colonizing ability of both VacA+ and VacA
H. pylori strains in CD1 mice, that only VacA+
H. pylori strains could provoke the infiltration
of inflammatory cells and proposed the VacA+ H. pylori SPM326-infected CD1 mouse model. Lee et al. (11) isolated a strain of H. pylori with high colonizing ability,
the Sydney strain, by screening fresh clinical H. pylori
isolates with BALB/c and SJL mice and suggested that the Sydney strain could become a standard strain of H. pylori for use in mouse
models. Marchetti et al. (12) and Lee et al. (11)
noted that their H. pylori-infected mouse models could be
used for the study of pathogenesis, the screening of novel therapeutic
agents, and the development of vaccines. However, in order to confirm
the contribution of IFN- to H. pylori-induced gastric
inflammation, it is necessary to develop new mouse models which can
conclusively define the contribution of this cytokine. In the present
study, we designed an H. pylori-infected
IFN-/ mouse model and investigated the role of
IFN- in protection from H. pylori infection and in the
development of gastric inflammation.
MATERIALS AND METHODS
Top
Abstract
Introduction
Materials and methods
Results
Discussion
References
/ and wild-type C57BL/6 mice
were used in this study. All of the mice were specific-pathogen-free
6-week-old males. The wild-type C57BL/6 mice were purchased from
Charles River Japan (Atsugi, Japan). IFN-/ mice were
generated by a targeted mutation of the IFN- gene as described
previously (18). Briefly, a nonfunctional IFN- gene was
created by inserting the -galactosidase gene and the neomycin
resistance gene at the translation initiation site in the first exon of
the IFN- gene. The targeting vector containing the disrupted IFN-
gene was transfected into A3-1 embryonic stem cells. G418-resistant
embryonic stem cells were selected and analyzed by PCR and Southern
blot hybridization to examine targeted disruption of the IFN- gene.
These cells were injected into blastocysts from superovulating C57BL/6
females, and then the embryos were transferred to the uteri of
pseudopregnant ICR recipients. The male chimeras were crossed with
C57BL/6 females, and then embryonic stem cell-derived pups were
backcrossed to C57BL/6 mice. The genotypes of the IFN- locus were
checked by Southern blot hybridization. Homozygous
IFN-/ mice were produced by intercrossing
heterozygous mice.
gene function of offspring mice was
verified by measuring IFN- mRNA in spleen cells from offspring mice
stimulated with phorbol myristate acetate and ionomycin and IFN-
protein in cultures of spleen cells from offspring mice treated in the
same manner. IFN- mRNA was detected by Northern blot hybridization
analysis. IFN- protein was measured by an enzyme-linked
immunosorbent assay (ELISA) (18).
/ mice and wild-type mice grew normally and were healthy.
Bacteria. The following H. pylori strains were used in this study. H. pylori ATCC 43504 and ATCC 51110 were obtained from the American Type Culture Collection (Rockville, Md.). H. pylori CPY2052, CPY3401, and HPK127 were kind gifts from M. Karita (Houfu Onsen Hospital, Houfu, Japan). H. pylori KP41a, KP48a, and KP64b were isolated from human gastric biopsy specimens at Kyoto Prefectural University of Medicine. All eight strains were gram-negative spiral rods and were positive for the H. pylori cagA and vacA genes.
Preparation of bacterial inocula.
Lyophilized ATCC 43504 and
ATCC 51110 were suspended in phosphate-buffered saline (PBS) (pH 7.4),
streaked on Helicobacter-selective brain heart infusion
(BHI) agar plates containing 7% (vol/vol) horse blood, vancomycin (10 µg/ml), polymyxin B (2.5 U/ml), trimethoprim (5 µg/ml), and
amphotericin B (2 µg/ml) (HSBHI agar plate) (Eiken Chemical Co.,
Tokyo, Japan), and incubated for 5 days at 37°C with an Aeropack
System charged with a gas mixture consisting of 80% N2,
15% CO2, and 5% O2 (Mitsubishi Gas Chemical
Co., Tokyo, Japan). One milliliter of PBS was added to each incubated
plate, and then the plate was rinsed. Two hundred microliters of the rinse solution was added to 5 ml of BHI broth containing 4% (vol/vol) fetal bovine serum (FBS) in a 20-ml test tube and cultivated for 24 h at 37°C under a gas mixture consisting of 80%
N2, 15% CO2, and 5% O2 on a
reciprocal shaker (200 rpm). One milliliter of FBS and 1 ml of glycerol
were added to this culture, and it was stored at 
80°C (stock culture).
Antigen preparation for oral immunization.
The inoculum
preparation of H. pylori CPY2052 described above was
centrifuged at 5,000 × g for 10 min. The pellet was washed three times with 20 ml of PBS (pH 7.4) by centrifugation, weighed as
wet matter, and suspended in PBS to obtain a 10% precipitate concentration (wet weight of precipitate/volume). The suspension was
sonicated twice for 10 min each time with an interval of 10 min by use
of an ultrasonic disrupter (Biorupter UCD 130; Tosho Denki Co.,
Yokohama, Japan) immersed in a melting ice bath. The protein content of
the bacterial sonicate was determined by the Lowry method, adjusted to
10 mg of protein/ml with PBS, and used as the antigen for oral
immunization and for an ELISA. The antigen preparations were stored at
80°C in aliquots until use.
Experimental protocol for H. pylori infection. All mice used in this study were challenged only once at the beginning of the experiment, after 8 h of fasting, with 0.5 ml of the inoculum preparation of H. pylori or PBS (pH 7.4) (as an uninfected control) by use of an oral stainless steel catheter. The mice were then maintained on the fast for 4 h and housed as described above. The colonization of gastric mucosa was assessed at specified times. Each experimental group comprised 6 to 10 mice.
Experimental protocol for oral immunization.
Wild-type and
IFN-/ mice were each divided into three groups and
administered one of the following three kinds of antigen solution:
group 1, 0.2 ml of PBS (pH 7.4) alone; group 2, 0.2 ml of PBS
containing 100 µl of the CPY2052 sonicate; and group 3, 0.2 ml of PBS
containing 100 µl of the CPY2052 sonicate plus 10 µg of cholera
toxin (Sigma Chemical Co., St. Louis, Mo.). Antigen administration was
carried out at days 0, 7, and 14, after 8 h of fasting, by use of
an oral stainless steel catheter. At day 21, all groups were challenged
with 0.5 ml of the inoculum preparation of CPY2052 after 8 h of
fasting. After the administration of the antigen solution or the
inoculum preparation, fasting was continued for 4 h. Colonization
of the gastric mucosa and the level of H. pylori-specific
serum immunoglobulin G (IgG) in each group were assessed at day 42. Each experimental group comprised 6 to 10 mice.
Microbiological evaluation. Mice were sacrificed at specified times, and their stomachs were resected. The resected stomachs were divided longitudinally into two halves and weighed. One half of each resected stomach was added to 1 ml of PBS and homogenized with a tissue grinder (Pellet Pestle Mixer; Kimble/Kontes). A 50-µl portion of the homogenized stomach was plated on an HSBHI agar plate and incubated for 5 days at 37°C with an Aeropack System as described above. The colonies on the plate were counted and expressed as CFU per gram of tissue.
For determination of whether the organisms of the colonies were H. pylori, a PCR was carried out with DNA prepared from the colonies. Five of the colonies on the plate were scratched and suspended in 1 ml of PBS. The suspension was centrifuged (10,000 × g for 5 min), and the precipitate was washed with 1 ml of PBS by centrifugation. The precipitate was resuspended in 50 µl of distilled water and boiled for 10 min. The supernatant obtained by centrifugation (10,000 × g for 5 min) was stored at20°C until use as a PCR template (bacterial DNA preparation).
Four oligonucleotide primers were designed based on the published
sequences of the H. pylori ureB and cagA genes
and synthesized with a DNA synthesizer (Gene Assembler Plus; Pharmacia
Biotech, Uppsala, Sweden) as described previously (21). The
primers for the ureB gene were 5'-TGGGATTAGCGAGTATGT-3'
(sense) and 5'-CCCATTTGACTCAATG-3' (antisense), and
the primers for the cagA gene were
5'-GATAACAGGCAAGCTTTTGAGG-3' (sense) and
5'-CTGCAAAAGATTGTTTGGCAGA-3' (antisense).
Thirty microliters of the bacterial DNA preparation was denatured by
heating at 95°C for 10 min and cooled on ice for 5 min. Five
microliters of this denatured DNA preparation was used in the PCR.
Fifty microliters of the reaction mixture consisted of 10 mM Tris-HCl
(pH 8.8), 0.1% Triton X-100, 50 mM KCl, 1.5 mM MgCl2, 20 µM 2'-deoxynucleotide 5'-phosphates (Pharmacia Biotech), 20 nM each
primer, 1.0 U of Taq DNA polymerase (Boehringer GmbH, Mannheim, Germany), and 5 µl of the denatured DNA preparation. Amplification was performed with an automated thermal cycler (TP-3000; Takara Shuzo Co., Kyoto, Japan) for 35 cycles, each of which consisted of 1 min at 95°C for denaturation, 1 min at 50°C for annealing, and
1 min at 72°C for extension. The final cycle included an extension step for 7 min at 72°C to ensure full extension of the product. Ten
microliters of each PCR product was analyzed by electrophoresis on a
1.5% agarose gel containing ethidium bromide, and the bands were
examined under UV light.
Histological evaluation. The other half of each resected stomach was used for the histological evaluation. Longitudinal sections from the gastroesophageal junction to the gastroduodenal junction were fixed in neutral buffered 10% formalin, embedded in paraffin, sectioned at 5 µm, and stained with hematoxylin and eosin. The degrees of inflammation were assessed by microscopic observation without knowledge of the experimental groups and was expressed as follows: none, normal appearance of scattered mononuclear cells on the lamina propria (the same degree as in uninfected control mice); mild, moderate infiltration of mononuclear cells in the lamina propria and the submucosa and no erosions in the epithelium; moderate, moderate infiltration of mononuclear cells in the lamina propria and the submucosa and erosions in some parts of the epithelium; and severe, severe infiltration of mononuclear cells in the lamina propria and the submucosa and erosions in many parts of the epithelium.
ELISA for H. pylori-specific serum IgG. Blood was obtained from mice before killing, and serum was collected. Each well of a 96-well microtiter plate was filled with 100 µl of 0.1 M carbonate buffer (pH 9.6) containing 0.1 µl of the antigen preparation of H. pylori CPY2052 described above, incubated overnight at 4°C, washed five times with PBS (pH 7.4) containing 0.05% (vol/vol) Tween 20 (PBS-Tween 20), and blocked with 100 µl of PBS-Tween 20 containing 2.5% (wt/vol) nonfat dry milk for 1 h at 37°C. The plate was then washed five times with PBS-Tween 20. One hundred microliters of serum samples diluted 1:10 with PBS-Tween 20 was added to each well. The plate was incubated for 1 h at 37°C and rinsed once with PBS-Tween 20. One hundred microliters of alkaline phosphatase-conjugated goat anti-mouse IgG (Cappel, Durham, N.C.) diluted 1:1,000 with PBS-Tween 20 was added to each well, and the plate was incubated for 1 h at 37°C. After the plate was rinsed again with PBS-Tween 20, 100 µl of 3.8 mM p-nitrophenyl phosphate as a substrate was added to each well and incubated for 30 min at room temperature. The optical density at 405 nm was measured with a microplate reader (Titertek Multiscan). Negative control serum was obtained from 13-week-old wild-type mice and included in every assay.
Spectrophotometric urease assay.
Urease activity in live
bacteria was measured spectrophotometrically by modifying the method of
Fauchère and Blaser (7). H. pylori strains
were cultivated for 30 h in BHI broth containing 4% (vol/vol) FBS
as described above and harvested by centrifugation at 5,000 × g for 10 min. The precipitate was washed three times with
PBS (pH 6.5) by centrifugation and suspended in PBS. The bacterial
concentration of the suspension was adjusted to 8 × 108 CFU/ml by measuring the optical density at 550 nm with
a Shimadzu UV-120-01 spectrophotometer. Fifty microliters of the
bacterial suspension was added to each well of a 96-well microtiter
plate, and the reaction was started with the addition of 50 µl of 300 mM urea in PBS containing 0.5% (wt/vol) phenol red. The reaction was
carried out at 37°C, and the optical density at 570 nm was measured
at 20 and 50 min after the start of the reaction with a microplate
reader (Titertek Multiscan). Urease activity was evaluated by
calculating the increment in the optical density and was expressed as
follows: , below 0.01; ±, from 0.01 to 0.03; +, from 0.03 to 0.30;
and ++, above 0.30.
Statistics. Colonization and serum IgG values were expressed as means ± standard errors. Sample means were compared by the Mann-Whitney U test. Infection rates were compared by the two-tailed Fisher exact probability test. A P value of <0.05 was considered significant.
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RESULTS |
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Top
Abstract
Introduction
Materials and methods
Results
Discussion
References
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Screening of H. pylori strains useful for mouse
experimentation.
To identify the H. pylori strains with
colonizing ability in the mouse stomach and to determine whether
IFN- is involved in protection from H. pylori infection,
we first examined the colonizing abilities of eight human clinical
H. pylori isolates by using C57BL/6 wild-type mice and
IFN-/ mice 4 weeks after infection (Table
1). Three strains (ATCC 43504, CPY2052,
and HPK127) colonized both wild-type and IFN-/ mice,
and the magnitudes of infection were considerably high. However, the
colonization values were somewhat lower in the wild-type mice than in
the IFN-/ mice (Table 1). Strain ATCC 51110 did not
colonize either mouse strain at all, and strain KP48a colonized
IFN-/ mice but not wild-type mice. The other three
strains (KP41a, KP64b, and CPY3401) colonized IFN-/
mice and partially colonized wild-type mice.
|
/ mice. The
gastric tissues from the wild-type mice infected with CPY2052 showed a
moderate infiltration of mononuclear cells in the submucosa and the
lamina propria, whereas the same tissues from the
IFN-/ mice infected with CPY2052 did not show any
infiltration. These findings indicate that only strain CPY2052 can
cause the infiltration of inflammatory cells in the presence of
IFN-.
|
/
mice, one of which has inflammatory ability (CPY2052) and the other of
which does not have inflammatory ability (ATCC 43504 and HPK127).
Long-term infection test for confirming gastric inflammation.
To examine the inflammatory features of the CPY2052-infected wild-type
mouse model, we carried out a long-term infection test to compare this
model with the CPY2052-infected IFN-/ mouse model
(Table 2 and Fig.
2).
|
|
/
mice, no inflammatory symptoms were observed, even 15 months after infection.
Protection from H. pylori infection by oral
immunization.
To determine whether IFN- is involved in
protection from H. pylori infection by vaccination, we
carried out an oral immunization protection test by using the
CPY2052-infected wild-type mouse model and the CPY2052-infected
IFN-/ mouse model (Table
3). The CPY2052 bacterial sonicate was
given orally to mice with and without cholera toxin, used as an
adjuvant. In a control study, PBS was given instead of the antigen.
Mice were then exposed to strain CPY2052 as described in Materials and
Methods. All mice that received PBS were infected. In mice treated with
the bacterial sonicate alone, protection from H. pylori
infection was incomplete in both mouse models. In contrast, complete
protection was achieved in wild-type mice immunized with the bacterial
sonicate and cholera toxin, and almost complete protection was obtained
even in IFN-/ mice immunized in the same manner.
|
on the activation of humoral immune
responses in the presence of oral immunization, we performed an
additional experiment. The levels of H. pylori-specific
serum IgG in both mouse models were determined as one of the indices of
the activation of humoral immune responses (Fig.
3). The bacterial sonicate alone induced
no increase in the levels of H. pylori-specific serum IgG,
whereas the addition of the adjuvant significantly increased the levels
of H. pylori-specific serum IgG in both models. However,
there was no significant difference in the levels of H. pylori-specific serum IgG between the wild-type mice and the IFN-/ mice.
|
), H. pylori
sonicate (
), or PBS (
). IgG values were assessed by
measuring the optical density at 405 nm. Results are expressed as
mean ± standard error of the mean for data obtained from 6 to 10 mice per group. a, significantly different (P < 0.05)
from values for the H. pylori sonicate group and the PBS
group.
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DISCUSSION |
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Top
Abstract
Introduction
Materials and methods
Results
Discussion
References
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It appears that there are differences in infective ability among
H. pylori strains, since Lee et al. obtained only one
H. pylori strain that could infect mice from among biopsy
specimens from 23 patients (11). It is not yet known what
factors regulate the infective ability of H. pylori. Eaton
et al. studied the urease-dependent colonization of gnotobiotic piglets
by H. pylori and reported that urease activity was essential
for the colonization of gnotobiotic piglets by H. pylori
(3, 4). It has therefore been suspected that the urease
activity of 8 H. pylori strains has a strong influence on
the infective ability of the bacteria. In the present study, strain
ATCC 51110 (which has no urease activity) was not able to colonize,
while strains ATCC 43504, CPY2052, and HPK127 (which have urease
activity) colonized both wild-type and IFN-/ mice.
However, strain KP48a, which has urease activity, was not able to
colonize wild-type mice. These results indicate that urease is a
necessary but not sufficient factor for the colonization of mice by
H. pylori. Other bacterial features, such as motility (5, 6) and adherence to epithelial surfaces (9, 14, 17), may influence infective ability.
There have been few reports regarding the protective effect of IFN-
against Helicobacter infection. Mohammadi et al. suggested that a protective role of IFN- against H. felis infection
was unlikely, because the neutralization of IFN- by anti-IFN-
antibody treatment had no effect on the magnitude of infection in their H. felis-infected mouse model (13).
On the contrary, in our study, the H. pylori strain (KP48a)
without the ability to colonize wild-type mice was able to colonize IFN-/ mice, clearly demonstrating the positive
contribution of IFN- to protection from H. pylori
infection. However, based on our finding that H. pylori ATCC
43504, CPY2052, and HPK127, all with strong infective ability, could
colonize wild-type mice despite the expression of IFN-, we consider
that the protection induced by IFN- is not so strong. The present
results showed that IFN- was not essential for protection by oral
immunization, because oral immunization with an H. pylori
sonicate and cholera toxin was similarly effective in
IFN-/ mice and wild-type mice. The protection
mechanisms induced by oral immunization are thought to be different
from that based on IFN-. After oral immunization with the H. pylori sonicate and cholera toxin, the H. pylori-specific serum IgG levels increased significantly. However,
the levels in the immunized wild-type mice were almost the same as
those in the immunized IFN-/ mice. These results
suggested that the protective effect induced by oral immunization is
based on humoral immunity. IFN- might exert a protective effect via
host defense systems, including cellular immunity and the activation of
macrophages and natural killer cells, rather than humoral immunity.
One of the most interesting current issues is how colonization by a noninvasive bacterium, H. pylori, causes gastritis. The vacuolization of epithelial cells by cytotoxin is suspected to be one of the causes. However, because no vacuoles were observed in the epithelium even 15 months after infection in the CPY2052-infected wild-type mouse model, although inflammatory responses were confirmed, it is not likely that vacuolization caused gastritis in this model.
We consider that IFN- is involved in the activation of mononuclear
cells, because the infiltration of mononuclear cells was observed in
CPY2052-infected wild-type mice but not CPY2052-infected IFN-/ mice. Mohammadi et al. reported that
anti-IFN- antibody treatment of mice caused a significant reduction
in in vitro IFN- production by splenic and gastric lymphocytes from
mice infected with H. felis and that this reduction in
IFN- production was accompanied by a reduction in the gastric
inflammation score (13); these results suggested that
IFN- was involved in the continuation of gastric inflammation. The
present study demonstrated the positive contribution of IFN- to
gastric inflammation caused by H. pylori infection.
Ye et al. reported that gastric epithelial cells constitutively
expressed the class II major histocompatibility complex and that the
constitutive expression of the class II major histocompatibility complex on gastric epithelial cells increased markedly during infection
with H. pylori. They also suggested that IFN- might enhance the antigen-presenting cell function of gastric epithelial cells through the up-regulation of the class II major
histocompatibility complex (22). The role of IFN-
indicated in the present study might involve intensification of the
antigen-presenting cell function of gastric epithelial cells.
Based on the present results, we conclude that IFN- may play an
important role in inflammation rather than protection from H. pylori infection, although IFN- is involved in both protection from H. pylori infection and the inflammation induced by
this infection. This study also shows that different H. pylori strains can cause various levels of infection or
inflammation in the presence or absence of IFN-.
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ACKNOWLEDGMENTS |
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We thank K. Kashima, Kyoto Prefectural University of Medicine, for kind advice and helpful discussions and T. Nishino, Kyoto Pharmaceutical University, for technical advice. We are also grateful to M. Karita for generously donating the H. pylori strains.
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FOOTNOTES |
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* Corresponding author. Mailing address: Third Department of Internal Medicine, Kyoto Prefectural University of Medicine, Kawaramachi, Hirokoji, Kamigyo-ku, Kyoto 602-0841, Japan. Phone: 81-75-251-5519. Fax: 81-75-251-0710. E-mail: sawai{at}basic.kpu-m.ac.jp.
Editor: R. N. Moore
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REFERENCES |
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Abstract
Introduction
Materials and methods
Results
Discussion
References
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| 1. | Bayerdörffer, E., N. Lehn, R. Hatz, G. A. Mannes, H. Oertel, T. Sauerbruch, and M. Stolte. 1992. Difference in expression of Helicobacter pylori gastritis in antrum and body. Gastroenterology 102:1575-1582[Medline]. |
| 2. |
Dalton, D. K.,
S. Pitts-Meek,
S. Keshav,
I. S. Figari,
A. Bradley, and T. A. Stewart.
1993.
Multiple defects of immune cell function in mice with disrupted interferon- genes.
Science
259:1739-1742 |
| 3. |
Eaton, K. A.,
C. L. Brooks,
D. R. Morgan, and S. Krakowka.
1991.
Essential role of urease in pathogenesis of gastritis induced by Helicobacter pylori in gnotobiotic piglets.
Infect. Immun.
59:2470-2475 |
| 4. |
Eaton, K. A., and S. Krakowka.
1994.
Effect of gastric pH on urease-dependent colonization of gnotobiotic piglets by Helicobacter pylori.
Infect. Immun.
62:3604-3607 |
| 5. |
Eaton, K. A.,
D. R. Morgan, and S. Krakowka.
1992.
Motility as a factor in the colonisation of gnotobiotic piglets by Helicobacter pylori.
J. Med. Microbiol.
37:123-127 |
| 6. | Eaton, K. A., S. Suerbaum, C. Josenhans, and S. Krakowka. 1996. Colonization of gnotobiotic piglets by Helicobacter pylori deficient in two flagellin genes. Infect. Immun. 64:2445-2448[Abstract]. |
| 7. | Fauchère, J.-L., and M. J. Blaser. 1990. Adherence of Helicobacter pylori cells and their surface components to HeLa cell membranes. Microb. Pathog. 9:427-439[Medline]. |
| 8. | Ghiara, P., M. Marchetti, M. J. Blaser, M. K. R. Tummuru, T. L. Cover, E. D. Segal, L. S. Tompkins, and R. Rappuoli. 1995. Role of the Helicobacter pylori virulence factors vacuolating cytotoxin, CagA, and urease in a mouse model of disease. Infect. Immun. 63:4154-4160[Abstract]. |
| 9. |
Goodwin, C. S.,
J. A. Armstrong, and B. J. Marshall.
1986.
Campylobacter pyloridis, gastritis and peptic ulceration.
J. Clin. Pathol.
39:353-365 |
| 10. | Kozol, R., A. Domanowsky, R. Jaszewsky, R. Czanko, B. McCurdy, M. Prasad, B. Fromm, and R. Calzada. 1991. Neutrophil chemotaxis in gastric mucosa, a signal-to-response comparison. Dig. Dis. Sci. 36:1277-1280[Medline]. |
| 11. | Lee, A., J. O'Rourke, M. Corazon de Ungria, B. Robertson, G. Daskalopoulos, and M. F. Dixon. 1997. A standardized mouse model of Helicobacter pylori infection: introducing the Sydney strain. Gastroenterology 112:1386-1397[Medline]. |
| 12. |
Marchetti, M.,
B. Aricò,
D. Burroni,
N. Figura,
R. Rappuoli, and P. Ghiara.
1995.
Development of a mouse model of Helicobacter pylori infection that mimics human disease.
Science
267:1655-1658 |
| 13. | Mohammadi, M., S. Czinn, R. Redline, and J. Nedrud. 1996. Helicobacter-specific cell-mediated immune responses display a predominant Th1 phenotype and promote a delayed-type hypersensitivity response in the stomachs of mice. J. Immunol. 156:4729-4738[Abstract]. |
| 14. |
Neman-Simha, V., and F. Mégraud.
1988.
In vitro model for Campylobacter pylori adherence properties.
Infect. Immun.
56:3329-3333 |
| 15. |
Nielsen, H., and L. P. Andersen.
1992.
Chemotactic activity of Helicobacter pylori sonicate for human polymorphonuclear leucocytes and monocytes.
Gut
33:738-742 |
| 16. | Nielsen, H., and L. P. Andersen. 1992. Activation of human phagocyte oxidative metabolism by Helicobacter pylori. Gastroenterology 103:1747-1753[Medline]. |
| 17. |
Segal, E. D.,
S. Falkow, and L. S. Tompkins.
1996.
Helicobacter pylori attachment to gastric cells induces cytoskeletal rearrangements and tyrosine phosphorylation of host cell proteins.
Proc. Natl. Acad. Sci. USA
93:1259-1264 |
| 18. |
Tagawa, Y.,
K. Sekikawa, and Y. Iwakura.
1997.
Suppression of concanavalin A-induced hepatitis in IFN-/ mice, but not in TNF- / mice: role for IFN- in activating apoptosis of hepatocytes.
J. Immunol.
159:1418-1428[Abstract].
|
| 19. | Tujii, M., S. Kawano, S. Tsuji, H. Fusamoto, T. Kamada, and N. Sato. 1992. Mechanism of gastric mucosal damage induced by ammonia. Gastroenterology 102:1881-1888[Medline]. |
| 20. | Yamaoka, Y., M. Kita, T. Kodama, N. Sawai, K. Kashima, and J. Imanishi. 1995. Expression of cytokine mRNA in gastric mucosa with Helicobacter pylori infection. Scand. J. Gastroenterol. 30:1153-1159[Medline]. |
| 21. | Yamaoka, Y., M. Kita, T. Kodama, N. Sawai, and J. Imanishi. 1996. Helicobacter pylori cagA gene and expression of cytokine messenger RNA in gastric mucosa. Gastroenterology 110:1744-1752[Medline]. |
| 22. | Ye, G., H. Gunasena, and V. E. Reyes. 1997. The role of the gastric epithelium in local T cell activation, p. 153-165. In P. B. Ernst, P. Michetti, and P. D. Smith (ed.), The immunobiology of H. pylori: from pathogenesis to prevention. Lippincott-Raven Publishers, Philadelphia, Pa. |
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