ORIgINAL PAPERS - Advances in Clinical and Experimental Medicine

ORIgINAL PAPERS - Advances in Clinical and Experimental Medicine

original papers
Adv Clin Exp Med 2013, 22, 3, 347–353
ISSN 1899–5276
© Copyright by Wroclaw Medical University
Iwona Radziejewska1, A–D, Małgorzata Borzym-Kluczyk2, B, C,
Katarzyna Leszczyńska3, B, C
Are Lewis b and H Type 1 on Helicobacter pylori
Involved in Binding of Bacteria to MUC1 Mucin?*
Czy struktury Lewis b i H typ 1 na Helicobacter pylori
uczestniczą w wiązaniu bakterii do mucyny MUC1?
Department of Medical Chemistry, Medical University of Białystok, Poland
Department of Pharmaceutical Biochemistry, Medical University of Białystok, Poland
3
Department of Microbiological Diagnostics, Medical University of Białystok, Poland
1
2
A – research concept and design; B – collection and/or assembly of data; C – data analysis and interpretation;
D – writing the article; E – critical revision of the article; F – final approval of article; G – other
Abstract
Background. Helicobacter pylori is responsible for gastroduodenal diseases such as chronic gastritis, gastric and
duodenal ulcers and also gastric malignances. During an infection, gastric mucins, especially secretory MUC5AC,
are said to participate in interactions with bacterial adhesins. Recently, epithelial MUC1 mucin has also been proposed to be engaged in infection development. H. pylori surface possesses lipopolisaccharides with Lewis antigens
whose role in interactions with gastric mucins has not been elucidated thoroughly so far.
Objectives. To check the involvement of Lewis b and H type 1 structures of H. pylori in adhesion of bacteria to
MUC1 mucin.
Material and Methods. The study was performed on gastric juices taken from 10 clinical patients and 4 H. pylori
strains. Bacteria were assessed for the presence of Lewis b and H type 1 structures by ELISA test. Interactions
of H. pylori with MUC1 were analyzed by sandwich ELISA method with bacteria pretreated or not with Lewis
b – HSA or H type 1 – HSA glycoconjugates.
Results. In the majority of the patients, the slight increase in adhesion of H. pylori to MUC1, after pretreatment of bacteria with H type 1 glycoconjugate was observed. A statistically significant difference was revealed in one strain with
a dose of conjugate 50 µg/mL (P = 0.05). The influence of Lewis b on adhesion is considered to be contradictory.
Conclusions. H type 1 antigens are suggested to be involved in carbohydrate – carbohydrate interactions of
H. pylori with MUC1 mucin (Adv Clin Exp Med 2013, 22, 3, 347–353).
Key words: Helicobacter pylori, H type 1, Lewis b, MUC1.
Streszczenie
Wprowadzenie. Helicobacter pylori jest bakterią odpowiedzialną za wiele chorób, w tym stany zapalne, wrzody
żołądka i dwunastnicy, a także nowotwory żołądka. W czasie zakażenia mucyny żołądkowe, a szczególnie mucyna
wydzielnicza MUC5AC, uczestniczą w oddziaływaniach z adhezynami bakterii. Ostatnio proponuje się także udział
mucyny błonowej MUC1 w rozwoju zakażenia. Na powierzchni H. pylori znajdują się lipopolisacharydy ze strukturami Lewis, których rola w oddziaływaniach z mucynami żołądkowymi nie została dotychczas wyjaśniona.
Cel pracy. Próba wyjaśnienia udziału struktur Lewis b i H typ 1 na H. pylori w adhezji bakterii do mucyny
MUC1.
Materiał i metody. Praca została przeprowadzona na sokach żołądkowych pobranych od 10 pacjentów i na 4 szczepach H. pylori. Obecność struktur Lewis b i H typ 1 na bakteriach oznaczono testem ELISA. Oddziaływania H. pylori z mucyną MUC1 analizowano testem sandwich ELISA z bakteriami traktowanymi lub nietraktowanymi strukturami Lewis b – HSA i H typ 1 – HSA.
*This work was supported by Medical University of Białystok grant 3-03504F.
I. Radziejewska, M. Borzym-Kluczyk, K. Leszczyńska
348
Wyniki. U większości pacjentów zaobserwowano nieznaczny wzrost adhezji H. pylori do mucyny MUC1 po wstępnym traktowaniu bakterii glikokonjugatami H typ 1 – HSA. Zmianę istotną statystycznie wykazano w przypadku
jednego badanego szczepu przy zastosowanej dawce konjugatu 50 µg/mL (P = 0.05). Wpływ struktury Lewis b na
adhezję wydaje się niejednoznaczny.
Wnioski. Antygen H typ 1 może uczestniczyć w oddziaływaniach typu węglowodan–węglowodan zachodzących
między H. pylori i mucyną MUC1 (Adv Clin Exp Med 2013, 22, 3, 347–353).
Słowa kluczowe: Helicobacter pylori, H typ 1, Lewis b, MUC1.
Helicobacter pylori colonizes the gastric mucosa of about half of the world’s population and
is responsible for gastroduodenal diseases such as
chronic gastritis, gastric and duodenal ulcers and
also gastric malignances [1–3]. It is interesting that
most infected individuals do not reveal any clinical
symptoms [4]. It seems to be clear that both bacterial virulence factors and host susceptibility features play a role in the development of infection.
An intricate relationship between Helicobacter
pylori and the stomach mucosa at the cellular
and molecular level is proposed [5, 6]. Two secretory mucins MUC5AC and MUC6 and epithelial
MUC1 are major mucins of gastric mucus. The secreted MUC5AC is limited to the foveolar epithelium, whereas the expression of secreted MUC6 is
restricted to the glands. The membrane associated
MUC1 is expressed mainly in foveolar cells and, to
a much lesser extent, in mucus glands [4, 6–9].
Helicobacter pylori colonizes the gastric mucosa by adhering to the mucous epithelial cells and
the mucous layer lining the epithelium [4, 10]. The
bacterium has adhesins responsible for recognizing specific carbohydrate structures. The best defined adhesins are the blood group binding adhesin (BabA) with an affinity to Lewis b and H type 1
antigens (Fig. 1) and sialic acid binding adhesin
(SabA) that binds sialyl dimeric Lewis x [11, 12].
Human Lewis antigens represent terminal modifications on mucins [4, 10]. MUC5AC mucin with
Lewis b structure seems to be the best proved receptor for Helicobacter pylori adhesins [6, 13, 14].
Recently, the involvement of MUC1 mucin in
the interaction with the bacterium has been proposed [8, 9].
Lewis blood antigens are also expressed on
the O-specific chains of the lipopolisaccharides
(LPS) of Helicobacter pylori. This phenomenon
can be understood as a kind of molecular mimicry between bacteria and host. This, in turn, may
result in immune tolerance against antigens of
the pathogen and can facilitate colonization [6, 8,
15]. Moreover, it is known that Helicobacter pylori strains are able to adapt their outer membrane
expression profile according to alterations in the
host environment, including changes in mucosal
glycosylation patterns, by switching on and off
specific gene expression [6, 8, 15]. However, the
importance of Lewis blood antigens on H. pylori
surface has not been elucidated thoroughly so far.
According to this the authors decided to check if
the mentioned structures could be involved in interactions with carbohydrates of MUC1. The main
aim of this study was to assess the effect of addition of Lewis b – HSA and H type 1 – HSA glycoconjugates to H. pylori on binding of bacterium to
MUC1 mucin. Many aspects of this subject should
be explored. The authors believe that their results
will contribute to an explanation of the problem.
Material and Methods
Patients and Specimens
Ten Helicobacter pylori infected patients with
duodenal ulcers (7 males and 3 females; aged 24–52;
mean age 40) hospitalized in the Department of
Medicine and Gastroenterology of Regional Hospital
of Białystok, Poland, were included in the study. The
patients were treated for 2 weeks with oral administration of omeprazole (2×20 mg per day), amoxiciline (2×100 mg) and tynidazole (2×500 mg). All the
subjects were on a standard hospital diet served to
peptic ulcer patients. The tested gastric juices were
taken on 11–13 day of the successful treatment. The
presence of the bacterium was examined histopatologically and by urease test with gastric cells scraped
under endoscopic examination.
To obtain high molecular mass material, the
juices were chromatographed on a Sepharose 4B
column as described before [18]. Concentrated
material of the void volume was subjected to further analysis. The protein content was measured
using bicinchoninic acid [19]. Samples of juices
were diluted to the same protein concentration
prior to ELISA tests.
Bacterial Strains
and Culture Conditions
Helicobacter pylori strains were isolated from
gastric epithelial cells scraped from four individuals suffering from gastritis. The scrapings were
collected before the beginning of the treatment,
under endoscopic examination, from the prepyloric area and the body of the stomach. The scrap-
Binding of Helicobacter pylori to MUC1
349
Fig. 1. H type 1 and Lewis b carbohydrate structures.
Abbreviations: Fuc, fucose; Gal, galactose; GlcNAc,
N-acetylglucosamine
Ryc. 1. Struktury cukrowe H typ 1 i Lewis b.
Skróty: Fuc, fukoza; Gal, galaktoza; GlcNAc,
N-acetyloglukozoamina
ings were immediately carried into the transport
medium Portagerm pylori (bioMerieux, France).
After homogenization, the bacteria were cultured
on Pylori Agar and Columbia Agar supplemented
with 5% sheep blood (bioMerieux, France) for
7 days at 37oC under microaerophilic conditions
using Genbag microaer (bioMerieux, France).
Microorganisms were identified upon the colony
morphology, by the Gram method; the activity of
the bacterial urease, catalase and oxidase were also
determined. To prove H. pylori species, ELISA test
(HpAg48; EQUIPAR, Spain) was used. Then the
bacteria were subcultured in the same conditions
and suspended at 1.2×109 bacteria/mL in PBS.
Determination of Lewis b,
H Type 1 Antigens and Adhesins
Directed to Lewis b and
H Type 1 Glycoconjugates
on H. pylori Strains
Lewis b and H type 1 expression on four H. pylori strains were measured by enzyme-linked immunosorbent assay (ELISA). H. pylori isolates were
diluted in phosphate-buffered saline (PBS) to a final
concentration ∼ 2.4 × 107 bacteria/mL. Then, 100 µL
of the bacterial suspension was coated onto microtiter
plates (NUNC F96; Maxisorp, Roskilde, Denmark)
and incubated at room temperature (RT) overnight.
The plates were washed 3 times (100 µL) with PBS,
pH 7.4, 0.05% Tween (PBS-T; washing buffer) between all ensuing steps. Unbound sites were blocked
with 100 µL of blocking reagent for ELISA (Roche
Diagnostics, Mannheim, Germany) at RT for 1h. In
order to check the presence of proper adhesins to
Le b and H type 1 structures, to the part of the wells
Fig. 2. Assessment of Lewis b and H type 1 structures
on Helicobacter pylori. Strains (1, 2, 3, 4) were not
affected (white columns) or were allowed to react with
Lewis b – HSA (grey columns) (A) or H type 1 – HSA
(black columns) (B) before the reaction with proper
antibody in ELISA test
Ryc. 2. Ocena struktur Lewis b i H typ 1 na
Helicobacter pylori. Szczepy (1, 2, 3, 4) nietraktowane żadnym czynnikiem (białe kolumny), traktowane
Lewis b – HSA (szare kolumny) (A) lub H typ 1 – HSA
(czarne kolumny) (B) przed reakcją z odpowiednim
przeciwciałem w teście ELISA
100 µL of Le b (Le b – HSA; 22 mol oligosaccharide/
mol HSA; 10 µg/mL of blocking reagent) or H type 1
(H type 1 – HSA; 25 mol oligosaccharide/mol HSA;
15 µg/mL) (IsoSep AB, Tullinge, Sweden) were added. The rest of the wells were left in blocking buffer.
All the plates were incubated 2h at RT and then they
were treated with primary antibodies, anti – Le b and
anti – H type 1 diluted in PBS-T-BSA (1%) (specifications of antibodies employed in this study are listed
in Table 1) (BSA – bovine serum albumin; Sigma, St
Luis, MO, USA) at RT for 1 h. Next, the plates were
incubated with a secondary antibody, biotin conjugated rabbit anti-mouse IgG, diluted in the above
buffer, at RT for 1h. Then the plates were incubated
with horseradish peroxidase (HRP) avidin D (Vector, Burlingame, CA, USA) at RT for 1 h. After being washed with PBS (4 times), the coloured reaction
was developed by incubation with 2,2’-azino-bis(3ethylbenzthiazoline-6-sulfonic acid) (ABTS) – liquid
substrate for horseradish peroxidase (Sigma, St Luis,
MO, USA). The absorbance was recorded at 405 nm
after 15–30 min.
I. Radziejewska, M. Borzym-Kluczyk, K. Leszczyńska
350
Binding of H. pylori
to MUC1 Mucin
The anti-MUC1 monoclonal antibody E 29
(diluted in 0.1M bicarbonate buffer, pH 9.5 to final concentration 0.5 µg/mL) was coated (100 µL/
well) onto microtiter plates overnight at 4oC. The
plates were washed 3 times (100 µL) in washing
buffer after each ensuing step. Unbound sites were
blocked as described above. The plates were incubated for 2 h with aliquots (50 µL; 5 µg of protein/
mL) of gastric juices (concentrated void volumes
after gel filtration) at RT. The bacteria were diluted
50 times with PBS-T-HSA (20 µg/mL) (HSA – albumin from human serum; Sigma, St Luis, MO,
USA) and incubated for 15 min in this solution.
Two structures: Le b – HSA or H type 1 – HSA
(5 or 50 µg/mL) were added to the part of bacterial
suspension 1h before the transfer to microtiter
plates. The plates were incubated with H. pylori,
H. pylori/Le b and H. pylori/H type 1 overnight
at 37oC. Then each well was treated with anti – H. pylori polyclonal, biotin-conjugated antibody
at RT for 1 h. After being incubated with HRP
avidin D, HRP activity was determined as described above.
Statistics
The results obtained from determination
of H. pylori (pretreated or not with Lewis b and
H type 1 structures) binding to MUC1 mucin were
subjected to statistical analysis (by STATISTICA
7.1 StatSoft program). To determine statistical
significance and type of distribution the authors
used ANOVA test and Kolmogorov-Smirnov test
for normality. To assess equality of variances in
examined groups, the Levene’s test was used. Post
hoc comparisons were based on NIR test. P values ≤ 0.05 were considered significant.
This study was approved by the Institutional
Ethical Committee with the principles of the Declaration of Helsinki and informed consent was obtained from all patients.
Results
Two of four examined H. pylori strains (strain
1 and 3) revealed relatively a high level of Le b antigens on their surface in comparison with strain
2 and 4 (Fig. 2A). The authors assumed that binding of Le b – HSA structures to bacteria means the
presence of proper adhesins on H. pylori surface.
So the increased level of examined antigens should
be observed. However the authors noticed a rather
slight increase of the expression of Le b structures
assessed after addition of Le b – HSA to the bacterial suspension. So they suppose that examined
strains possess a very low level of adhesins directed
to Le b receptors. Expression of H type 1 structure
was low in all examined H. pylori strains (Fig. 2
B). Small amount of adhesins directed to H type 1
antigen could be suggested only on strain 1.
Fig. 3. Lewis b addition effect on Helicobacter pylori binding to MUC1 mucin (n = 10) selectively captured by anti
MUC1 antibody. H. pylori was allowed to react with Lewis b – HSA prior to addition to mucin. White columns – H. pylori without pretreatment; grey columns – H. pylori treated with 5µg/mL of Le b – HSA; black columns – H. pylori treated with 50 µg/mL of Le b – HSA. Binding of H. pylori without pretreatment with Lewis b – HSA is
stated as 100%. The bars represent mean ± SD
Fig. 3. Rezultat dodania struktury Lewis b na wiązanie Helicobacter pylori do mucyny MUC1 (n = 10) selektywnie
związanej przez przeciwciało anty MUC1. H. pylori reagowała z Lewis b – HSA przed dodaniem do mucyny. Białe
kolumny – H. pylori nietraktowana żadnym czynnikiem; szare kolumny – H. pylori traktowana Le b – HSA w stężeniu
5 µg/mL; czarne kolumny – H. pylori traktowana Le b – HSA w stężeniu 50 µg/mL. Wiązanie H. pylori do mucyny bez
dodawania Lewis b – HSA ustalono jako 100%. Kolumny przedstawiają średnie ± SD
Binding of Helicobacter pylori to MUC1
351
Fig. 4. H type 1 addition effect on Helicobacter pylori binding to MUC1 mucin (n = 10) selectively captured by anti
MUC 1 antibody. H. pylori was allowed to react with H type 1 – HSA prior to addition to mucin. White columns
– H. pylori without pretreatment; grey columns – H. pylori treated with 5 µg/mL of H type 1 – HSA; black columns
– H. pylori treated with 50 µg/ml of H type 1 – HSA. Binding of H. pylori without pretreatment with H type 1 – HSA
is stated as 100%. The bars represent mean ± SD; p ≤ 0.05
Fig. 4. Rezultat dodania struktury H typ 1 na wiązanie Helicobacter pylori do mucyny MUC1 (n = 10) selektywnie
związanej przez przeciwciało anty MUC1. H. pylori reagowała z H typ 1 – HSA przed dodaniem do mucyny. Białe
kolumny – H. pylori nie traktowana żadnym czynnikiem; szare kolumny – H. pylori traktowana H typ 1 – HSA w stężeniu 5 µg/mL; czarne kolumny – H. pylori traktowana H typ 1 – HSA w stężeniu 50 µg/mL. Wiązanie H. pylori do
mucyny bez dodawania H typ 1 – HSA ustalono jako 100%. Kolumny przedstawiają średnie ± SD; p ≤ 0.05
The addition of a higher dose (50 µg/mL) of Le
b – HSA glycoconjugate to H. pylori caused a slight
increase of adhesion to MUC1 mucin in three of examined strains (strain 1 by 2%; strain 2 by 4.5% and
strain 4 by 4%) (Fig. 3). With a smaller dose (5 µg/
mL) such an increase was observed only in two
strains (strain 2 by 3%; strain 4 by 0.2%). In strain
1 and 3, a dose of 5 µg/mL revealed an inhibition effect on binding. In strain 3 also a higher dose of Le
b – HSA caused the same effect but smaller than the
effect observed with a dose of 5 µg/mL. However all
these changes were not statistically significant.
In the case of H type 1 structure, slight inhibition effect was observed only in strain 3 with
a dose of 5 µg/mL (Fig 4). In the rest of the strains,
addition of H type 1 – HSA structures caused dose
dependent increase of binding of the bacterium to
MUC1 mucin. However, these increases were very
subtle (strain 1 with a dose of 5 µg/mL by 4% and
with a dose of 50 µg/mL by 10%; strain 2 by 6 and
14%; strain 4 by 5 and 6% respectively). For strain
3 an increase of binding was revealed only with
higher dose of H type 1 – HSA by 5%. Statistically
significant increase was revealed only in strain
2 with a dose of 50 µg/mL (P = 0.05).
Discussion
To colonize the human stomach, Helicobacter
pylori interacts with glycan structures of the host
glycocalyx [20, 21]. Mucins as the main compo-
nents of gastric mucus are likely to participate in
these interactions. It is accepted that MUC5AC
possesses sugar antigens which act as receptors for
Helicobacter pylori adhesins [6, 11]. Recently also
glycoforms of MUC1 mucin are taken into account
as potential receptors for the bacterium [4, 6, 13].
Many models of interactions between mucins and
the bacterium have been proposed [14]. One of
them involves H. pylori lipopolisaccharides (LPS)
which share structural similarity to Lewis blood
group antigens in gastric mucosa and are said to
participate in colonization. Such a bacterial molecular mimicry could provide an escape for H.
pylori from the host humoral response by preventing the formation of antibodies shared by the host
and bacterium [8, 10, 15, 22, 23]. However, the role
of these antigens of H. pylori surface is not thoroughly understood.
In this study the authors decided to check what
will be the effect of addition of Le b and H type 1
glycoforms to H. pylori on binding with MUC1
mucin. They assumed that the mentioned structures would be bound to specific adhesins on the
bacterium surface and in this way “block” the potential sites which could be involved in the interactions with carbohydrates of mucins. Surprisingly
present results showed slight increase of binding,
especially for H type 1 antigen. In case of Lewis b,
the results seem to be rather contradictory.
Upon present observations the authors can
hypothesize that carbohydrate – carbohydrate in-
I. Radziejewska, M. Borzym-Kluczyk, K. Leszczyńska
352
Table 1. Source of antibodies used in this study
Tabela 1. Źródło przeciwciał stosowanych w badaniu
Antibody (Przeciwciało)
Clone (Klon)
Source (Źródło)
Final dilution
(Ostateczne
rozcieńczenie)
Anti-MUC1
E 29
Thermo Scientific
1:400
Anti-Lewis b
LWB01
Abcam
1:800
Anti-H type 1
17-206
Abcam
1:500
Anti-H. pylori (polyclonal, biotin-conjugated)
Abcam
1:2000
Biotin-conjugated rabbit mouse-IgG (whole molecule)
Sigma
1:5000
teractions may be involved in mucin – bacteria relations. Their results can suggest involvement of
H type 1 glycoforms of H. pylori in such interplays
with MUC1 mucin. Carbohydrate – carbohydrate
interactions are known as a novel and highly versatile mechanism for cell adhesion and recognition. Carbohydrate self recognition can take place
through surfaces determined by the carbohydrate
epitopes and is based on noncovalent bonds: van
der Waals contacts, hydrogen bonds, hydrophilic
and hydrophobic interactions [24]. It is likely that,
in the present study, in the majority of samples,
H type 1 glycoconjugate added could enhance
bacterial aggregation by carbohydrate – carbohydrate interactions and facilitate adhesion to mucin
receptors. It was proposed that bacterial aggregation can be promoted by, for example, anti – Le
x monoclonal antibodies which can mediate Le
x – Le x interactions [15]. However, direct carbohydrate – carbohydrate interactions cannot be
excluded.
Present results indicate that, in some cases,
small doses of Le b or H type 1 structures could
inhibit binding of H. pylori to mucin, probably by
blocking of proper adhesins on bacteria. However,
the authors revealed that there is really a very low
expression of examined adhesins on the bacterial
surface. So it can explain rather small inhibition
effect involving adhesins.
It is rather difficult to explain why in some
cases no effect on adhesion of H. pylori to mucin after pretreatment of bacteria with Le b or
H type 1 structures has been observed. It is known
that many other kinds of interplays are possible to
be involved simultaneously, different structures
on both bacteria and the host can be implicated.
Some carbohydrate structures of H. pylori can be
involved in mutual interactions occurring between
bacteria.
Summing up, present results can suggest the
existence of carbohydrate-carbohydrate interactions between H. pylori lipopolisaccharides and
carbohydrates of MUC1. The authors propose
that H type 1 glycoform can be involved in such
interplays. However, they want to emphasize that
this conclusion should be treated very carefully.
This study is preliminary and more detailed investigations are needed to prove or disprove the proposed idea. Finding structures, which are involved
in adhesion of bacteria to mucin, may contribute
to looking for new therapeutic strategies of drugresistance H. pylori infections.
Acknowledgments. The authors thank Professor Z. Namiot from Department of Medicine and Gastroenterology,
Regional Hospital of Białystok, Poland for kindly providing them with the gastric juices.
References
[1] Eslick GD: Helicobacter pylori infection causes gastric cancer? A review of the epidemiological, meta-analytic, and
experimental evidence. World J Gastroenterol 2006, 12, 2991–2999.
[2] Uemura N, Okamoto S, Yamamoto S, et al.: Helicobacter pylori infection and the development of gastric cancer.
N Engl J Med 2001, 345, 784–789.
[3] Wang C, Yuan Y, Hunt RH: The association between Helicobacter pylori and early gastric cancer: a meta-analysis.
Am J Gastroenterol 2007, 102, 1789–1798.
[4] Magalhaes A, Reis CA: Helicobacter pylori adhesion to gastric epithelial cells is mediated by glycan receptors. Braz
J Med Biol Res 2010, 43, 611–618.
[5] Amieva MR, El-Omar EM: Host-bacterial interactions in Helicobacter pylori infection. Gastroenterology 2008,
134, 306–323.
[6] Linden S, Mahdavi J, Hedenbro J, et al.: Effects of pH on Helicobacter pylori binding to human gastric mucins;
identification of binding to non-MUC5AC mucins. Biochem J 2004, 384, 263–270.
Binding of Helicobacter pylori to MUC1
353
[7] Gendler SJ: MUC1, the renaissance molecule. J Mammary Gland Biol Neoplasia 2001, 6, 339–353.
[8] Kobayashi M, Lee H, Nakayama J, et al.: Roles of gastric mucin-type O-glycans in the pathogenesis of Helicobacter
pylori infection. Glycobiology 2009, 19, 453–461.
[9] Nordman H, Davies JR, Lindell G, et al.: Gastric MUC5AC and MUC6 are large oligomeric mucins that differ in
size, glycosylation and tissue distribution. Biochem J 2002, 364, 191–200.
[10] Moran AP, Gupta A, Joshi L: Sweet-talk: role of host glycosylation in bacterial pathogenesis of the gastrointestinal
tract. Gut 2011, 60, 1412–1425.
[11] Ilver D, Arnqvist A, Őrgen J, et al.: Helicobacter pylori adhesin binding fucosylated histo-blood group antigens
revealed by retagging. Science 1988, 279, 373–377.
[12] Mahdavi J, Sonden B, Hurtig M, et al.: Helicobacter pylori SabA adhesin in persistent infection and chronic
inflammation. Science 2002, 297, 573–578.
[13] Linden SK, Sheng YH, Every AL, et al.: MUC1 limits Helicobacter pylori infection both by steric hindrance and
by acting as a releasable decoy. PLoS Pathog 2009, 5 e1000617. doi:10.1371/journal.ppat.1000617.
[14] Linden SK, Wickstrom C, Lindell G, et al.: Four models of adhesion are used during Helicobacter pylori binding
to human mucins in the oral and gastric niches. Helicobacter 2008, 13, 81–93.
[15] Sheu SM, Sheu BS, Yang HB, et al.: Anti-Lewis X antibody promotes Helicobacter pylori adhesion to gastric epithelial cells. Infect Immun 2007, 75, 2661–2667.
[16] Backstrom A, Lundberg C, Kersulyte D, et al.: Metastability of Helicobacter pylori bab adhesin genes and dynamics in Lewis b antigen binding. Proc Natl Acad Sci USA 2004, 101, 16923–16928.
[17] Solnick JV, Hansen LM, Salama NR, et al.: Modifications of Helicobacter pylori outer membrane protein expression during experimental infection of Resus macaques. Proc Natl Acad Sci USA 2004, 101, 2106–2111.
[18] Radziejewska I, Borzym-Kluczyk M, Kisiel DG, et al.: The effect of Helicobacter pylori eradication treatment
on the MUC1 and Lewis antigens level in human gastric juice: a preliminary study. Dig Dis Sci 2008, 53, 2641–
2645.
[19] Smith PK, Krohn RJ, Hermanson GT, et al.: Measurement of protein using bicinchoninic acid. Anal Biochem
1985, 150, 76–85.
[20] Corfield AP, Carrol D, Myersough N, et al.: Mucins in the gastrointestinal tract in health and disease. Front
Biosci 2001, 6, D1321–57.
[21] Hooper LV, Gordon JL: Glycans as legislators of host-microbial interactions: spanning the spectrum from symbiosis to pathogenicity. Glycobiology 2001, 11, 1R–10R.
[22] Appelmelk BJ, Monteiro MA, Martin SL, et al.: Why Helicobacter pylori has Lewis antigens. Trends Microbiol
2000, 12, 565–570.
[23] Moran AP, Sturegard E, Sjunneson H, et al.: The relationship between O-chain expression and colonization
ability of Helicobacter pylori in a mouse model. FEMS Immunol Med Microbiol 2000, 29, 263–270.
[24] Bucior I, Burger MM: Carbohydrate–carbohydrate interactions in cell recognition. Curr Opin Struct Biol 2004,
14, 631–637.
Address for correspondence:
Iwona Radziejewska
Medical University of Białystok
Department of Medical Chemistry
Mickiewicza 2a
15-222 Białystok 8
Poland
E-mail: [email protected]
Conflict of interest: None declared
Received: 27.02.2012
Revised: 17.04.2013
Accepted: 13.06.2013