0022-1554/90/$3.30
Thejournal ©
Copyright
Vol.
of Histochemistry and Cytochemistry 1990 by The Histochemical Society,
Received
ofLegal for
Medicine,
publication
NISHI,
MITSURU
Nara Medical
December
18,
1989
University,
Kashihara
and in revised
form
the distribution ofblood group-related antian indirect immunoperoxidase method with monoclonal antibodies (MAb) directed to A, B, H, Lewis a (Lea), Lewis b (kb), x (Le’), and Lewis y (LeY) antigem and Type 1 precursor chain in human pancreas. Effects ofprior digestion with exoglycosidases on MAb stainings were simultaneously investigated. A, B, H, b, and LeY antigens were detected in acinar cells and interlobular duct cells but not in centroacinar cells, intercalated duct cells, and islet of Langerhans cells. The expression ofthese antigens in acinar cells was not dependent on Lewis type and secretor status of the tissue donors, whereas that in interlobular duct cells was strictly dependent on secretor status. The distribution pattern of these antigens in acinar cells was not homogeneous, i.e., cells producing H antigens expressed both Leb and LeY antigens but not A or B antigens, whereas those producing A or B antigens did not seaete Leb and LeY as well as H antigens. Digestion with a-N-acetylgalactosaminidare or a-galactosidase resulted in the appearance of Lb and Le antigens as well as H antigen in acinar cells producing A and/or B antigens. Type 1 precursor chain was not de-
Introduction group-related
using creas
monoclonal antigens
analysis several
have
with
recently
of carbohydrate histochemical
to localize blood (10,13,20,21,31,32,36-39),
concerned
antibodies
malignant
(MAb)
been
directed
developed
and
to blood used
April
4, 1990;
group-related antigens in human panmost ofthese studies were mainly and
effects
ofthe
Lewis
type
and
accepted
it is necessary
gens
in normal In a series
ing method procedures of blood
and
submandibular
studies
the
3-based control
against To whom
correspondence
should
be addressed.
to know
distribution
and
fucosylation of the
tam
oligosaccharide MAb
combined
developed and
analyze
(14-19).
Type
antigens
of the anti-
of the Type secretor
(Se)
2, and
exact
gene.
in human
results
of these
(8,11),
glycosidase
strucpancreas previous
O-glycosidically in pancreatic chain
Because have
staindiges-
chemical
chains
3 precursor
antigens
structures with
The
1, Type
the lectin
endoglycosidase
the
are secreted
group-related
with
exo-
carbohydrate
glands
that
we have
with
to localize
ABH
blood
the precise
studies,
group-related
suggested
understand
I
(9A1862).
tissues. ofprevious
tures
that
in malignant
1990
in combination
tion
and
glycosylation
6,
in pancreatic tissues from reactors but appeared in cells producing H antigen after a-L-fucosidase digestion, which also disclosed Lea but not Lea antigen in acinar cells expressing both jb and LeY. In some non-reactors, MAb against Type 1 precursor chain reacted with acinar cells without enzyme digestion. Although Lea antigen was not detected in acinar cells, it was found in centroacinar cells, intercalated duct cells, and interlobular duct cells from all individuals examined except two Lc(a- b-) reactors. After sialidase digestion, Le antigen appeared in centroacinar and intercalated duct cells from some individuals. Sialidase digestion also elicited reactivity with MAb against Type 1 precursor chain in islet ofLangerhans cells from some individuals. These results demonstrate the complexity in the pattern of expression and regulation of blood group-related antigens in different cell types ofhuman pancreas. Such complexity may largely be ascribed to differences in individual genotypes and in gene expression patterns ofdifferent cell types. (J Histochem Cytochem 38:1331-1340, 1990) KEY WORDS: Blood group antigen; Immunohistochemistry; Monoclonal antibody; Exoglycosidase; Pancreas.
Type
ofaberrant
April
tected
secretor status of individuals on the expression of these antigens have not yet been fully explored in normal pancreatic tissues. To the mechanism
OKAMURA,
acinar
for
chains in different tissues (11). studies have been carried out by
tissues,
YOSHIRO
Nara 634, Japan.
tissues,
well-defined
structural Although
Artide
NAKAJIMA,
We examined gem using
Many
1990
USA.
Localization and Analysis of Blood Antigens in Human Pancreas Using with Monoclonal Antibodies and Digestion
NOBUAKI ITO,1 KATSUJI and TADAOMI HIROTA Department
1331-1340,
Printedin
Original
Histochemical Group-related Immunostaining Exoglycosidase
No. 9, pp.
38,
Inc.
rigid
linked acinar
is in part
well-defined specificity
cells under MAb
for cer-
an immunostaining
method
digestion
may also
procedure
1331
Downloaded from jhc.sagepub.com at VIRGINIA COMMONWEALTH UNIV on March 13, 2015
1332
ITO,
provide
a useful
bohydrate
means
chains
in
In the present using
several
nation
with
definite
tion
and
and
10%
study,
MAb
blood
formalin
filter
paper,
tissue
in human
MAb-H MAbLea
autopsy
formalin,
cases.
They
embedded
were fixed
in paraffin,
in
sections
were
incubated
with
MAb
MAbLex
at 0C
for
MAb-LeY
Donor
group,
sex, Blood
no.
and
age
Group
of the
tissue
donors” Age (years) 27
0
Le(a-b+)
S
M
0
Le(a-b-)
S
F
0
46E
0
Le(a
) NS
M
15
Le(a+b-)
73
+b
-
73E
0
NS
M
821
A Le(a-b+)
S
M
875
A Le(a-b+)
S
M
876
A
S
F
881
A Le(a+b-)
NS
M
7E
A
S
M
46
A
S
M
50
S
M
1
S
M
0
Le(a-b+)
A2 Le(a-b+)
37 9 months
4 0
747F
B
772
B
S
F
843
B
Le(a+b-)
NS
M
944
B
Le(a+b+)
S
M
S
M
89
S
M
20
S S
F M
35
AB
703 848
AB
850
AB
853
AB 4
I
Gal-U3 1-4)-GlcNAcl3Gal-(1-3)-GlcNAc-
a Gal, D-galactose; N-acetyl-D-glucosamine.
Le(a-b+) Le(a-b-)
S. secretor;NS. non-secretor; M. male; F. female.
GalNAc,
N-acetyl-D-galactosamine;
Fuc,
GIcNAc.
L-fucose;
nex; Dublin, CA). Finally, the sections were counterstained with hematoxylin, dehydrated, and mounted in balsam. A mouse MAb (1gM class) against the Trypanosoma surface antigen (a gift from Dr. Y. Takahashi, Department ofParasitology, Nara Medical University)was used as a negative control. As thehistochemical positive and negative controls for MAbLea, .jb Le5 -Le, and -Type 1, reactivity of these antibodies with the following tissue sites were examined: stomach surface mucosac from secretors (positive with MAbLeb and negative with MAb-Le5) and non-secretors (positive with MAb-Le5 and negative with MAbLeb); Brunncr’s gland (positive with MAbLeX and Le and negative with MAb-Le5 and Leb) (27); and proximal tubule and Henle’s loop of kidney (positive with MAb-Le5 and -Type 1) (8). These tissues were obtained from autopsy cases. Staining of sections from B and 0 donors with MAb-A or from A and 0 donors with MAb-B was also carried out as negative built-in control for MAb-A and -B. a-Galactosidase(green coffee beans)was purchased from Sigma(St Louis, MO); a-N-acetylgalactosaminidase (Acremonium sp.) was from Seitetsu (Tokyo,
Japan);
a-L-fucosidase
(Indianapolis,
(bovine IN),
and
kidney)
was
neuraminidase
from
Boeh-
(Arthmbacter
was obtained from Marukinshoyu (Tokyo, Japan). Digestion with a-galactosidase was carried out by incubating tissue sections in a solution containing the enzyme at a concentration of 2-2.5 U/mI in 3.2 M (NFLg)2SO4 solution, pH 6.0, at 37’C for 20-24 hr (19). Digestion with a-N-acetylgalactosaminidase and a-L-fucosidase was similarly performed by incubating sections in a solution containing the enzyme at a concentration of 1 U/mI in 0.1 M citrate buffer, pH 4.5 (17), and in 0.1 M citrate phosphate buffer, pH 5.2 (18), respectively, for 20 hr at 37’C. Digestion with neuraminidase was carried out by immersing sections with enzyme buffer solution (0.25 U/mI ofthe enzyme in 0.1 M acetate buffer, pH 5.2) for 6-24 hr at 37C (14). Sequential digestions ofsections were carried out by using the same incubation conditions described above for each enzyme. After the first enzyme digestion, sections were washed with distilled water and then incubated for a second enzyme digestion. ureafaciens)
Sex
845
M3
I Gafl3-
Fuc-(al-3) / Gal-(31-3)-GlcNAc3-
1
ringer-Mannheim
832
80E
in
Gal-(131-4)-GlcNAcl3Fuc-(al-3) / Gal-(31-4)-GlcNAcl3-
Fuc-(al-2)-
MAb-Type
Kogyo 1 . Blood
used
MAb
Fuc-(al-4) / Gal-(31-3)-GlcNAc3-
Fuc-(al-2)-
and
4 hr. MAb were used at dilution of i:so-i:io with PBS-BSA. Tissue sites reactive with MAb were visualized with the streptavidin-biotin-peroxidase complex method (4), using a StreptAvidin Immunostaining Kit (BioGe-
Table
by
GalNAc-(a1-3)-Gal3Fuc-(al-2) Gal-(al-3)Fuc-(al-2) Fuc-(al-2)-
distribu-
pancreas.
defined
HIROTA
Fuc-(al-4) from
neutralized
tissue
structures
OKAMURA,
study’
MAb-B
more
Methods
the
Determinant
MAb-A
sectioned serially at 4 tm. Typing ofdonor blood groups for ABO and Lewis was performed by routine hemagglutination testing of the donor’s blood samples. Secretor status of the donors was determined by the Lewis blood type or was histochemically deduced from the presence (secretor) or absence (non-secretor) ofH antigen in serous cells ofcorresponding submandibular glands (16). The blood samples and submandibular glands were also obtained from autopsy cases. The blood group ABO, Lewis, secretor status, sex, and age of the tissue donor are summarized in Table 1. MAb directed to A, B, and H antigens (MAb-A, MAb-B, and MAb-H) were purchased from Dako (Santa Barbara, CA). MAb against Le and Leb antigens (MAbLea and MAbLeb) were from BioCarb Chemicals (Lund, Sweden) or Signet Laboratories (Cambridge, MA), and MAb against Lex, Le, and precursor Type 1 chain (MAbLex, MAb-Le, and MAb-Type 1) were also from Signet Laboratories. The antigenic structures defined by these MAb are illustrated in Table 2. Tissue sections were dc-waxed, hydrated, and incubated in 0.1 M phosphate buffer, pH 7.2, containing 0.2 M NaCI (PBS), three times (each for 3 mm) at room temperature, and then incubated in PBS containing 1 mg/mI of bovine serum albumin (PBS-BSA) for 20 ruin. After being blotted dry with
2.
MAbLeb
were obtained
or 10%
Table
NAKAJIMA,
in combi-
to provide
as to the
antigens
method
antigens
procedures
information
of these
and
of car-
an immunostaining
group-related
digestion
comprehensive
tissues
structure
thepresent
we developed
against
regulation
the chemical
sections.
exoglycosidase
Materials Pancreatic
for analyzing tissue
NISHI,
6 months 51 1
month
50
Results Distribution
ofABH,
Immunoperoxidase antigens
showed
in acinar
cells
staining that
Le”, with
and MAb
A, B, H, Leb,
but not in centroacinar
Downloaded from jhc.sagepub.com at VIRGINIA COMMONWEALTH UNIV on March 13, 2015
Ley Antigens against
and
blood
LeY antigens
cells,
intercalated
group-related are secreted duct
cells,
I
.
,
. .f. .
s
.
‘-.-..,
,.
-p
S
‘
,
.
.
.
:s;;-:i
. ,,
.
t, ..
-
-.
.
.
‘%‘:IIl
.
-
,,i
.
. .
.
...
1-I
-
‘
S;;.
;,4_
.
/-
.
.i
-
--: .
,
:.-
M
.
:‘
.
t,
..‘
4
,
-,
(-
,.,
..I
.
.
l’d.:a4”
lB 11 -t.
.
4’ .
I
‘-
,
.
w
‘-,
.,
‘e
..-
-.
.;4aT
.
.
.;,
.t.
: ‘
‘
,
.
.,.
t ..
.
.
‘..
.
‘,
-
-
-
--. -
.
. .
.
:.4.
.
‘.Y
:
4.
:
-
I
.
;
-
-
4
I
.c..
.-,.
%
,4’.
I
,:::;
t_
#{149}-;.‘‘
,
.,-
. - ..
#{149}b
..,
-.
S
..
.
.‘4’
. ,
.
.
-,
1
..-
I’
‘
.
f#{149}
C
-
‘
:..‘
‘4 p
‘
i.,).
3A
r
‘q
4
L3B
11
.b4.
4,,
Figure 1. Sections of pancreas from a blood group AG secretor individual (donor and MAb-A- or -B-unreactive cells are stained with both MAbLeb and MAb-LeY.
no. 848) stained with (A) MAb-H, Bars = 100 tm.
Y
(B) MAbLeb,
and (C) MAb-LeY.
MAb-H-reactive
Figure 2. Sections of pancreas from a blood group AG secretor (donor no. 703) stained with MAb.Lea. (A) Nearly all the centroacinar cells and intercalated duct cells are stained with MAb-Le8. (B) Strong surface and cytoplasmic staining is seen at higher magnification. Interlobular duct cells are also stained with MAb-Le5. Bars: A = 200 tm; B = 50 tm.
a
Figure not react Figure
Bar
4. =
Sections of pancreas from a blood group at all with the pancreatic tissue, whereas Section
100 tm.
of pancreas
from
a blood
group
0, Le(a-b-) secretor individual MAb.Leb reacts with the acinar A non-secretor
(donor
no. 881)
(donor no. 845) stained cells. Bars - 200 tm.
stained
with
MAb-Type
with
1. Some
Downloaded from jhc.sagepub.com at VIRGINIA COMMONWEALTH UNIV on March 13, 2015
(A) MAb.Lea
acinar
cells
and (B) MAbLeb.
are
slightly
MAb-Le8
or moderately
does
stained.
1334
ITO,
, .
.
-nt
OKAMURA,
HIROTA
1%’
=,.
.“-‘
,..
:-..j.
.4 .
..c
.
.,,
-‘
.
-
.
NAKAJIMA,
l.,,t_..
#{149}) ::
#{149}
NISHI,
.
;
‘-
,
: -
.
4H. ,-.
5A
-
.
2.-
f’
‘f ‘
-
_t
I
.
,J
,.
?
:
.
:-
.
a
,-
‘
-,
.
.
,‘4*
..
:
.
#{149}
r
.
...
.,.
,
.
...
.
0
d’
.
..-;,a-
‘6B
..:
.,,-,
-f,.
‘#{149}c4
L .1
#{149}‘
Figure 5. Sections of pancreas from a blood group B secretor individual (donor no. 772) stained with (A) MAbLeb and (B) MAbLeb after u-galactosidase digestion. MAbLeb reacts with acinar cells expressing H antigen but not with those expressing B antigen. as shown in Figure 1. After enzyme digestion, MAbLeb reacts with nearly all the acinar cells, including those expressing B antigen. Islet of Langerhans cells (arrowheads) are not stained by MAbLeb before or after enzyme digestion. Bars = 200 rim. 6.
Figure
Sections
tion. Without after enzyme
and
of pancreas
islet of Langerhans stained
by
cells
weakly
with
reacted
not
distribution
Figure
of
homogeneous.
observed
7. Sections
the
acinar
of pancreas
digestion. After Bars = 100 m.
a small
-B, -H,
cells
producing
pattern
of
from
a blood
digestion,
and
(donor
of cells
interlobular
expressing
enzyme
A secretor
number
only
MAb-A,
cells
group
Although
MAb.Leh,
A reverse
between
fucosidase digestion.
=
a blood
only a limited 200 rim.
cells.
ubiquitously
The
from
enzyme treatment, digestion. Bars
duct
antigens
distribution
AB
MAb-Type
and
secretor
with
H antigen
but
not
verse
was
a mosaic
(donor
Leb,
Nearly
and
antigens.
both
Leb
and
cell
u-N-acetylgalactosaminidase
and
their
clusters
cells
diges-
are reactive
Thus,
the
Le’
antigens
cell
distribution
producing
with
MAb-LeY
clusters
express-
(Figures
1A-1C)
was
exactly
the
A or B antigen.
re-
Such
pattern of antigen distribution was observed in all the AB individuals examined, although the number of cells
stained
expressing
of the
after
all the acinar
Le’
produce
of that
no. 848)
(B) MAb-Le
A or B antigen,
A, B, and
with a cell cluster
and
MAb-LeY.
H,
ing
was
those
individual
1 reacts
(A) MAb-LeY
expressing
these
-Le’.
A or B antigen
group
of
with
are reactive
cells were
number
these
antigen
no. 7E) stained
or cell clusters
with
(A) MAb-H,
H antigen.
(B) MAb-Type
No reactivity
with
1, and
MAb-Type
(C) MAb-Type
1 is seen
without
1 after
cz-L-
enzyme
Figure 8. Sections of pancreas from a blood group B secretor (donor no. 944) stained with (A) MAb-Le’ and (B) MAb-Le5 after a-L-fucosidase digestion. No reactivity is seen withoutthe enzyme digestion. After the enzyme digestion, reactivity with MAb-Le’ appears in the supranuclear region of some acinar cells. Bars = 50 tm. Figure
9.
intercalated
Section
duct
of pancreas
cells,
from
a blood
and interlobular
duct
group
cells
B secretor
are reactive
individual
with
Figure 10. Section of pancreas from a blood group A secretor (donor are stained with MAb-Type 1 after enzyme digestion. Bar = 100 tm.
(donor
MAbLex
no. 944) stained
after
enzyme
no. 821) stained
with
with
MAbLex
digestion.
Bar
MAb-Type
1 after
after =
100
neuraminidase
neuraminidase
Downloaded from jhc.sagepub.com at VIRGINIA COMMONWEALTH UNIV on March 13, 2015
digestion.
Centroacinar
cells,
m.
digestion.
Islet cells of Langerhans
BLOOD
GROUP
ANTIGENS
IN
HUMAN
1335
PANCREAS
--. .
.-,
-t
l
,
.
-.
t .;,
.
-
..-
-..t
7A
7B
e.a.
;_
-. a
I
b
,.
,,#{149}‘
,
.,
-
-w
st..
H-o..
.
‘II
tp,..o ,.
0 .
‘
#{149} .
-
.:-
c
... -
-5’
.
-‘
.,.
,
‘
.
..,
‘‘
#{149}
.‘
S
e
‘jr
--
j
‘
T:
:,‘!
8tB1,,:
“at
;“:
4
4
.#{231}c., “.
,
#{149}.1 .
4_
.
.p-#r
,-
.-_‘
::
k$f:.
.
-
, ya.b
-
“e;
..
;b
.a-.-.
....
I
;
0
.
a “C-
#{149}
..eI
-e
_s.t
,
5ep
l
,.
#{149}
.-,
‘I
#{149}0
_..
a
#{149}‘
---,c
-p
I
.
q”up
,,
fri
4O1
(1
e
,
..
-‘-::
-1
S
.
:#{149}
:
#{149}
‘ .-
%
7’C;
‘
Downloaded from jhc.sagepub.com at VIRGINIA COMMONWEALTH UNIV on March 13, 2015
4 .
.:,
..
ITO,
1336
expressing
H, Leb, and
als. In half
of the
of cells gens,
produced
in others
cells
Leb,
and
and
nearly
LeY antigens
small
among
examined,
in all the
was relatively
0 individuals,
was variable
B individuals
H, Leb,
whereas
ofthese
LeY antigens
A and
not
than
MAb
the number
10%).
cells were
30-50%
In blood
stained
Leb and Ley antigens region of acinar cells,
cated
in the cytoplasm
gens
were
also
with
The expression of the secretor detected
at the
of these antigens status of the tissue
in interlobular
MAb-A,
-B, and
cells from
fine
duct
-H reacted
corresponding
granular
lateral
gen
cells
from
red blood
blood
ABH
anti-
of acinar
cells.
in acinar cells was independent donors; these antigens were not
with group
cells and endothelial Le antigen
from
any
blood
Distribution
Type
Le-’,
Precursor
reacted
dividuals,
with
whereas
a small
number
of acinar
in others
observed. Such individual group ABO and secretor was not detected
with MAbLea tercalated duct
no reactivity
with
variability was not status of the donors.
in acinar
cells,
cells
antibody
or strong
of centroacinar of interlobular
WAS
ever, staining tion with
these
ofSialidase
Sialidase
digestion
in-
nar cells and
was
uals
examined
secretors some
acinar
(Figure
cells from
some
4). In one blood
centroacinar
and
stained by MAb-Type with red blood cells
(Figures
shown).
3A and
MAb-Type
reactivity
tion. ABO
cells, induct cells
elicited reactivity 10). Such effects
non-secretors group
intercalated
but not those
AB, Lu (aduct
3B) and
1 occasionally
cells
react
resulted
cells, intercalated affected by enzyme
reactivity
in slight
fucosidase
acinar
Pre-existing
diges-
and in the perinuclear
after
with
with
sequential
fucosidase MAbLeb appeared
Le antigen not
the reHow-
reactivity
diges-
cells
reactivity
duct cells, digestion.
after with
and
enMAb-
interlobular
Digestion reactivity duct
(Figure
with
cells from
9). In some
cells reacted
cases,
with
MAb-Le’ some only
MAb-Le’
in centroaci-
but not all individa limited after
Such individual variability was not related and secretor status of the donors. Sialidase
number
enzyme
diges-
to the blood group digestion likewise
with MAb-Type 1 in islet ofLangerhans cells (Figure ofsialidase were also observed in a limited numIn one blood
with
MAb-Type
group
0, Le (a-
1 appeared
b
-)
at the surface
secretor,
weak
of most
of the
centroacinar cells and intercalated duct cells after enzyme digestion_ In other individuals, only a small number of these cells were with the MAb results obtained findings
after enzyme in this study
digestion. are summarized
as to the distribution
b-)
individual,
sor chains
(14,15)
moderately
included
in l#{224}ble3.
and
Type
3-based
ofType ABH
in Table
2 and Type
antigens
3.
3 precur-
(17-19)
are also
LeY antigens
in aci-
1. MAbLex, Lea, and -Type 1 did not react and endothelial cells from any blood group
Discussion Although
of a-galactosidase
and
a-N-acetylgalactosaminidase Although producing
Digestion
H, Leb, and Le antigens A or B antigen, prior
a-galactosidase
digestion
both
and
ing
with
not shown).
were
examined.
Effects
anti-
MAb-Type
a-N-acetylgalactos-
cells after
did
revealed
duct
Previous
with
cells
not
with
individuals,
Digestion
intercalated
of intercalated
from
reacted
in acinar (data
other
MAb-H
with
In some
intensity with with MAbLex
(data
Effects
stained The
cells
with
reactivity
in any acinar
cells expressing
digestion
in-
observed
enzymes.
8), MAbLea
MAb-Le
duct
reactivity
the
digestion
weak.
not appear
of acinar (Figure
ber ofindividuals.
o5
Sequential
was usually 1 did
MAb-Type
throughout the glands from all individuals examined (Figures 2A and 2B), except for one blood group 0 and one blood group AB, Lu (a - b - ) secretor individual. In these two cases, Lea antigen could not be detected in any cells, whereas reactivity with MAbLeb and terlobular
the
unmasked
7A-7C).
Lea in centroacinar duct cells was not
related to the blood Although Lea anti-
moderate
observed at the surface cells, and in the cytoplasm
in some
the
reduced
simultaneously
1 (Figures
zyme
1 Chain
MAb-Le’
gen
and
with
aminidase or a-galactosidase and a-L-fucosidase imparted activity with MAb-Type 1 in cells expressing A or B antigen.
tion
group.
ofLe”,
and
region
was usu-
ally detected on red blood cells only from blood group 0 donors, while Leb antigen was not found on red blood cells or endothelial cells
and pattern
Digestion
digestion
reduction of staining Although reactivity
non-secretors.
donors.
ofa-L-fucosidase
a-L-fucosidase
in the were lo-
form.
membrane
intensity
HIROTA
used.
Effects
MAb-H,
were mainly distributed whereas ABH antigens
in a diffuse,
detected
the staining
OKAMURA,
group
and
Usually, perinuclear
did not affect
NAKAJIMA,
A or B anti-
AB individuals
(less
all the acinar
but
enzymes
individu-
about
NISHI,
MAbLeb
A or B antigens
B, or AB individuals
revealed
MAbLey,
or eliminated
or intensified as MAb-H,
(Figures
5 and
6). Red
likewise
reacted with stained
with these only
a-N-acetylgalactosaminidase the reactivity
with
in cells blood
MAb-H enzymes. by MAb-H
cells and
with
producfrom LeY,
A, but
or -B, respectively.
reduced These
of A, B, H, Leb,
appears not to be under the of the Type 1 precursor chain
secretors
suggests
part that
under the fucosylation
chain cells
On the other after enzyme
or a-galactosidase MAb-A
in cells or
the reactivity
as well
not with Leb, after digestion hand, endothelial cells were digestion.
could not be detected a-N-acetylgalactosaminidase
the expression
nar cells presence
that
fucosylation
of the Type
control of the Se gene. of the O-glycosidically
is also under
the
control
and
control of the in non-secretors
of the
Previous linked
Se gene, the but not in
1 chain
may
be in
studies suggested Type 3 precursor
Se gene
in pancreatic
as a regulator
gene
acinar
(17,19). The
Se gene
the expression
has been ofthe
synthesis (41). Recently, model which postulates ent a-2-L-fucosyltransferase. gene expressed mainly
regarded
H gene,
which
codes
controlling
for a-2-L-fucosyltransferase
Oriol et al. (29) proposed a new genetic that the H and Se gene codes for a differAccording to their in tissues of mesodermic
Downloaded from jhc.sagepub.com at VIRGINIA COMMONWEALTH UNIV on March 13, 2015
hypothesis, the H origin encodes an
BLOOD
GROUP
Table
3.
ANTIGENS
Blood
grou
IN
HUMAN
p-related
1337
PANCREAS
antigens
dete
cted
in
human
Acinar cells
Antigen ABH
(Type
pancreas
Centroacinar
t heir
and
distribution”
and
intercalated
duct
Interlobular
cells
duct
cells
+
-
-
I ;- (S),
-
ABH (Type
2)
+
-
-
I
+
(5),
- (NS)
ABH
3)
+
-
-
I
+
(5),
-
-
+
+
-
-1+
-
Lea Leb Le’
Endothelial
cells
-
-
-
+
-
-
-
-
(NS)
1)
(Type
Islet
cells
(NS)
+
+(S),-(NS) -
+(S),-(NS)
-
-
-
-
LeY
+
-
ALeb,BLeb
+
-
-
-
-
ALeY,BLeY
+
-
-
-
-
SialylLea
-
+
-
-
Sialyl-Le’
-
+
-
-
Type
+
I
-
+
-(S),+(NS)
1 PR
Type2PR Type
-
+
-(S),+(NS)
3 PR
-
-
-
+
-
-1+
-
-
-
+
-
Sialyl-Type
1 PR
-
Sialyl-Type
2 PR
-
-
-
+
+
Sialyl-Type
3 PR
+
+
+
+
+
a
S,
b
Positive
secretor;
NS. in
non-secretor;
one
Le(a
a-2-L-fucosyltransferase whereas the Se gene This
cent
different
that
reacts
gene
studies
present
suming
and
that
ferases
are
- I
no.
+
antigens
.
which
previous
both
the H and in acinar
with
(17,19) and
occurs
in the
absence
reacts preferentially with these The expression of Leb antigens
positive
non-secretors
could
the H gene-specified In fact,
also
enzyme
the H gene-specified
the Type
1 and
Type
by as-
H gene-specified
although of these
Se gene-specified
precursors. in acinar
cells
from
by the
presence
1 precursor
a-2-L-fucosyltransferase
Lewisof
can use both
substances
duct
(28).
to be likewise the
Se gene
In these
under (28).
tissues
the control In contrast
cells, the expression dependent on the
the expression
(2,33).
of H antigen
of the products to acinar
cells,
of both
Ex-
the
Se gene-specified
fucosyltransferase
the
al-2
of precursors.
fucosylation
is believed The
the H and
to function
enzymes
In agreement munohistochemical
encoded with
by the previous
studies
(39),
A, B, and lectin
but
not others.
pattern
of distribution
im-
producing
H, Leb,
Ley antigens
suggested
Erythrina
finity
for
Consistent
and
with
LeY antigens
the
in pancreatic
existence
those
acinar
cells
of SBA-reactive
ofSBA
The
2 precursor
which
not
to Type results
with
Gal-
pancreas
3 precursor of the
BLeb,
a high
with
acinar
producing
afcells
A and/or
quite
B
well with
for the a-GalNAc
irrespective
with
we found
has
residues
by SBA (14) but not by ECA.
1 and Type
obtained
which
correspond
except
be attnibuta-
study,
reacted
those
sites ofECA
digestion
ALeY,
(9,40),
can be recognized
appeared
a-galactosidase
(ALeb,
but
SBA may
(ECA),
chain
the binding in human
antigen,
with
In a preliminary
agglutinin
H antigen
In contrast
chain.
cristagalli
Thus,
son chain
the reactivity
2 precursor
Type
producing
ofA
digestion,
Type
prior
chains, secretor
the Type 2 precurstatus
of the
donor.
a-N-acetylgalactosaminidase
suggest
that
BLeY) are produced
difucosyl
A and
in acinan
cells
or B antigens
along
with
the monofucosyl Type 1, Type 2, and Type 3-based A and B antigens. The presence of difucosyl A and B antigens carried by glycosphingolipids definite
(5).
has been However,
information
disclosed Because
(15) and
study.
Let’, and LeY antigens may be regarded but not Lea and Lex antigens, is the
of Leb and
fucosidase to the
intestine
activity of than that
in the present
in cells
studies
that
Se genes.
histochemical
a mosaic
in
appearance
of Lea and Leb antigens in these cells suggests that the the Le gene-specified enzyme is much more prominent of the
duct
to be strictly hence, only
preferential
after
seems
in interlobular
of blood group antigens appears secretor status of the donors and,
precursor
antigens.
chains.
pression of Leb antigen in non-secretors has been reported in unnary epithelium (30), deep glands ofdigestive mucosa (24,35), and sweat
individuals
(1-3,4)-GlcNAc in acinarcells expressing H antigen (14). Because MAb-Type 1 reacted with acinar cells expressing H antigen only ble
enzyme
was observed
Le’ antigen
Previous
a-2-L-fucosyltransthe
as acceptor
some
and the coexpression ofH, as evidence that H antigen, (26,41).
can use Type
2 precursors
of cells from
antigens
Lea and
direct
be explained
be explained
that
1
can
that
of the
number
these results, the pattern of distribution of Leb and LeY antigen is likewise the reverse of that of A or B antigen. The absence of
in different tissues gene model, the results
Se gene-specified cells
in a small
of ABH
by releast two
enzyme can use the Type 1 and Type 3 precursor chains, its affinity for these precursors is low and the accumulation that
detected
were
the Type
has been supported the presence ofat
showed
studies
expressed
precursors
+
853).
preferentially
model
types of a-2-L-fucosyltransferase On the basis ofthis two-structure
(2,23,25).
ofthe
chain;
(donor
I
reacting preferentially with Type 2 chains, expressed in epithelial tissues encodes a different
two-structure
biochemical
precursor
b - ) individual
-
a-2-L-fucosyltransferase chain.
PR.
-
about
by the the
that
present
the ALeb
and
the
tissue
histochemical once
transferase
BLeb structures
Downloaded from jhc.sagepub.com at VIRGINIA COMMONWEALTH UNIV on March 13, 2015
in human
biochemical
Leb structure,
the A or B gene-specified that
reported
pancreas
study origin
could of these
and small not
provide
antigens,
as
an acceptor
for
study. formed, (22,34),
is not
it has been
are constructed
presumed
by the fucosyla-
1338
ITO,
lion
ofthe
C-4 position
gen (41). The
ofGlcNAc
process
ofthe
is catalized
1-based
A or B anti-
by a-4-L-fucosyltransferase
Type
coded
antigen
appear
not
ing is in contrast
NISHI,
NAKAJIMA,
to be hydrolyzed
to the fact
OKAMURA,
HIROTA
by the fucosidase.
that
Fuc residues
This
of Type
find-
1, 2 H anti-
by the Le gene. A similar mechanism may be operating in the synthesis of ALey and BLeY structures (26). In this case, it is presumed
gens and Le antigen are hydrolyzed by the enzyme in histochemical systems as disclosed in the present study as well as a previous
that
one
Type
2-based
fucosylated
A or B antigens
the ALeX and A or B antigen
and
were
not
sized
in the manner
detected,
the Type
1 and
precursor
because these
ture
and
difucosyl
and
then
by the X gene
cells
may
(41) and Lloyd
A or B antigen
may
are
to give
serve
be synthe-
digestion
the biochemical
is compatible
expressing
with
their
LeY antigen
substrates
systems,
and
Le or X gene
struc-
acceptor
can
a-4-Luse
substrates
Type
in these
1 and
cells.
the presence of sialylated Lea antigen tnoacinar and ductular cells.
Table
4.
Chemical
Precursor
structures
terminal
of
blood
of Lea and Le’ an(1). In the present
Fuc residues
group-related
of the
antigens
the
Leb
found
wA
observed
whether present
Because
in
human
Uchida
the Le (a
-
b
et al. (39)
-
coded
2 precursor
strong
and
reactivity
Itai
by the
chains with
as
MAb-
et al. (13) reported
on the
) individual
membrane
is presumed
of cen-
to lack
the
Le
(Type
2)
(Type
2)
pancreas Gal31-3GlcNAc-
(Type
1). Gal31-4GlcNAc3-
Ga131-3GalNAcal-Ser/Thr H antigen
beprevi-
sialidase digestion, it is difficult to ascerLea antigen is produced in these cells in
without
sialylated study.
Type
Because
antigenic structures, tigen are only slowly
however,
different
as reported
or a-3-L-fucosyltransferase
the
tam
systems,
the Fuc residues
is somewhat
histochemical
Lea
whereas those by the enzyme
used
their
and
in this
respectively, hydrolyzed
of the fucosidase
to hydrolyze
to the stenic effect of al-4 Fuc It is notable that the ability
ously (14,16,18). The preferential expression of Lea, x and their sialylated forms in centroacinar cells and intercalated duct cells means that only
Thus,
as the direct
chemical
enzyme
study. In biochemical systems, the Fuc residues of both Lob and Ley antigens are hydrolyzed by the fucosidase, yielding Lea and Le’
histochemical
specificity
to attack
sialyltransferase
in cells
ofthe
be attributable of Leb antigen.
biochemical
tween
A or B antigen. of Lex antigen
inability
of the enzymes
Le antigens (26).
(18). The
ofLeb antigen may residues on GIcNAc
of difucosyl monofucosyl
Leb and
determinants
by Watkins
Type 2-based
appearance
fucosidase
in these
proposed
of difucosyl
after
produced
coded
BLeY structures. Because the expression is strictly confined to cells producing
A or B antigen,
The
are first
by cz-3-L-fucosyltransferase
(Type
Gal3l.-3GlcNAc3-
(Type
Gall31-4GlcNAc3-
1),
1-2
1-2
Fuca Gal3
3)
Fuccz I
-3GalNAca
1 -Ser/Thr
(Type
3)
1-2
Fuca A(B) antigen
GaINAc(Gal)ct
1 -3Gal
1-3GIcN
Ac
(Type
I)
(Type
2)
1-2
Fucts GaINAc(Gal)a
I -3Gal3
1 -4GIcNA4I 1-2
Fuca GalNAc(Gal)a
1-3Gal
1-3GalNAca I -2
Fuca
Le4 antigen
Gal
I -3GlcNAcl1-4 Fuca
Leb antigen
Ga113 1 -3GlcNAcl1-2
Fuca A(B)Leb
antigen
GaINAc(Gal)a
1 -3Gal
l-3GlcNAc1-2
Fucct Le” antigen
1-4
Fuca
Gal5
1-4
Fucci I -4GlcNAc1-3
Fucu
Le antigen
Gal3
l-4GlcNAc31-2
Fuc
A(B)LeY antigen
GalNAc(Gal)a
1-3Gal3
1-4GlcNAc31-2
Fuca
Downloaded from jhc.sagepub.com at VIRGINIA COMMONWEALTH UNIV on March 13, 2015
1-3
Fuc 1-3
Fucci
1 -Ser/The
(Type
3)
BLOOD
GROUP
:
ANTIGENS
Centroacinar
and duct
HUMAN
IN
-
intercalated
cell
- --
.
i-
-
A(type
1-
Le,
.X.?ee
Precu,so,
-
precursor
(type
Langerhans
1,
,/Le
-
Lea
-
1.
type
1,
Literature
group-related in Table
antigens
found
in human
4.
1.
type
2)-it.Leb.
jB
2(
B(type
1.
Le’
K, McKibbinJM,
2. Betteridge A, Watkins a-2-L-fucosyltransferase 13:1126, 1985
gene
type
Cited
Hakomori 5: The monoclonal antibody to difucosylated type 2 chain (Fuca1-2Gal1-3[Fuca1-3]GlcNAc; determinant). J Biol Chem 258:11793, 1983
1. Abe
gene
gene
type
of blood
are presented
ALeS’
;
7(
jSe
H(type
l)-,I.ALCb.
IA gene Se
..__...H(type
gene
type
J
H,
structures
Le,xgene
Le
.
Sialylated
ical
pancreas
.
H LCa.
1339
PANCREAS
2)BLeb.
directed Y
WM: Acceptor substrate specificities from different tissues. Biochem
of human
Soc Trans
BLe
3. Bj#{246}rk S, Breimer jLe.
Leb.
11. Proposed
in different
cell
-
4.
duct
scheme
types
gene
Le’
(nter(obu(ar
Figure
X
ME, Hansson GC, Karlsson K-A, Leffler H: Structunes of blood group glycosphingolipids of human small intestine. A relationship between the expression offucolipids ofepithelial cells and ABO, Le and Se phenotype ofthe donor.) Biol Chem 262:6758, 1987
cell
for the biosynthesis
of human
pancreas.
of blood-group
In centroacinar
and
ABH antigens intercalated
duct
cells and interlobular duct cells, Type 2-based Le#{176} and Le antigens are minor components or not expressed, whereas Type 1-based Lea and Leb antigens are consistently expressed. Le#{176} antigen is usually detected in its sialylated form in centroacinar
gene
and
and
fails
Lea antigen
cult
form
with
blood
the
presence
of Leb in urinary
of an
Orntoft
with
a-4-L-fucsyltransferase
individual,
that
the
and
antigens,
such
reported
the
urothelium
Lewis
type
8. Cordon-Cardo
C, Lloyde
9.
Dc Boeck
H, Loontiens
10.
of an of erythro-
12. Franqois B, Oriol
Sialylated
rule
out
not correspond
Type
1 precursor
of Langerhans
cells
from
the possibility to that chains
some
that
the Lewis
ofpancreatic
are likewise
individuals.
type
acinar detected
Sialidase
cells.
digestion soybean obsenva-
tions).
recognize
3 precur-
Because
these
lectins
the Type
2 or Type
tic forms by sialylation. The results obtained in the present distribution pattern of blood group human differences
pancreas.
Such
complexity
in the individual
expression ofdifferent cell types. The ways of blood group-related antigens pression
in different
cell types
may
largely and
that complex
14.
the in
be explained
in the pattern
by
of gene
proposed biosynthetic pathand the patterns of gene cx-
are illustrated
in Figure
11. The
them-
in
individu-
Finstad
CL,
McGroarty
ME,
Reuter
VE,
FG, Lis H, Sharon
N: Binding
ofsimple
carbo-
oligosaccharides
mdi-
with a fluorescent 1984
by monoclonal and glycolipids 1985
antibodies that carbohydrate are onco-developmental an-
A, Sansonetti N, Mollicone R, Le Pendu), GaltonJ,)anulmes R: Heterogeneity of Lewis antibodies. A comparison of the ofhuman and animal 50:227, 1986
Itai 5, Arii
5, Tobe
R, Kitahara
reagents
with
synthetic
A, Kim
Y, Yamabe
oligosaccharides.
H, Ohtsuki H, Kin2-6 sialylation 1988
hara Y, Shigeta K, Kannagi R: Significance of 2-3 and of Lewis a antigen in pancreas cancer. Cancer 61:775,
chains, i.e., to their cryp-
study demonstrate antigens is very
genotypes
Vox Sang 13.
cells with peanut agglutinin, and Ito et aL, unpublished
son chain (9,16,17), three different types of precursor Type 1, 2, and 3 chains, are produced and converted
reaction
in islet
elicited the reactivity ofislet agglutinin, and ECA (14,40;
B human
Ernst C, Thurin J, Atkinson B, Wurzel H, Herlyn M, Stromberg N, Civin C, Koprowski H: Monoclonal antibody localization of A and B isoantigens in normal and malignant fixed human tissues. Am) Pathol 117:451, 1984
antigens
does
KO,
hydrates and some N-acetyl-Iactosamine-containing to Erythrina cristagalli agglutinin as followed cator ligand. Arch Biochem Biophys 234:297,
1 1. Feizi T: Demonstration structures of glycoproteins tigens. Nature 314:53,
(8), we cannot
as expressed
A and
Bander NH, Old U, Melamed MR: Immunoanatomic distribution of blood group antigens in the human urinary tract. Influence of secretor status. Lab Invest 55:444, 1986
cyte is not responsible for that of all the tissues and, hence, not for the true genotype of individuals. Because MAbLca and MAbLeb used in this study are highly specific for their corresponding oferythrocyte
group 1984
H, Levery SB, McKibbinJM, Hakomori S: Blood group A dewith monoand difucosyl type 1 chain in human erythrocyte membranes. Biochemistry 24:3578, 1985
may
related
of glycosphingolipids ofblood 228:71,
7. Clausen terminants
in-
however,
specificity intestine Biophys
6. Brodin T, Chester MA, Karlsson K-A, Messeter L, Zopf D, Lundblad A: A monoclonal antibody that recognizes both Leb and Y (LeY) antigens. Glycoconjugate J 4:399, 1987
aberrant
antigen
ME: Tissue
pancreas and small als. Arch Biochem
its
com-
has been (3), and red
ofLeb
in the
suggesting
cells
Recently,
individuals intestine
et al. (30),
are
5. Breimer
it is diffi-
in acinar
occurrence
of
and
cells
However,
antigen
of the MAbLeb
(6,12,24).
absence
chain
duct
) individuals.
-
an unexpected
to crossreactions
Le (a-b-)
b
-
1 precursor
individual.
of Leb
Le (a
the
intercalated
antigen in Le (a-b-) epithelium (30), small
Such
antigens
presence
of Type and
of this
cells from
cells (7).
be due as Le
presence
the genotype
duct
occurrence reported
the
a-4-L-fucosyltransfenase,
in centroacinar
to explain
terlobular
cells.
to produce and
sialylated
patible
ductular
Bonnard C, Papermaster DS, Kraehenbyhk R: The streptavidin-biotin bridge technique: application in light and electron microscope immunocytochemistry. In Pollak J, Varndell IM, eds. Immunolabelling for electron microscopy. Amsterdam, Elsevier, 1984, 95
15.
Ito N, Nishi K, Nakajima M, Ishitani A, Okamura Y, Matsuda Y, Hirota T: Histochemical reactivity ofsoybean agglutinin with blood group antigens and their precursor substances in acinar cells ofhuman pancreas. J Histochem Cytochem 35:881, 1987 Ito N, Nishi K, Nakajima M, Matsuda Y, Ishitani A, MizumotoJ, Hirota Localization of blood group antigens in human pancreas with 1cctin-horseradish peroxidase conjugates. Acta Histochem Cytochem 19:205, 1986
T;
16. Ito N, Nishi K, Nakajima M, Okamura Y, Hirota ‘F Histochemical ysis ofthe chemical structure ofblood group-related carbohydrate in serous
cells
and glycosidase
of human
digestion.
Downloaded from jhc.sagepub.com at VIRGINIA COMMONWEALTH UNIV on March 13, 2015
submandibular ) Histochem
glands using lectin Cytochem 37:1115,
anal-
chains staining 1989
1340
17.
ITO,
Ito N, Nishi
K, Nakajima
M, Okamura
Y, Hirota
T Histochemical
demonstration ofO-glycosidically linked, type 3 based ABH antigens in human pancreas using lectin staining and glycosidase digestion proce-
dunes. 18.
Histochemistry
92:307,
1989
Ito N, Nishi K, Nakajima M, Okamura Y, Hirota T: Effects of a-Lfucosidase digestion on lectin staining in human pancreas. ) Histochem Cytochem 36:503, 1988
19. Ito, Nishi K, Nakajima idase digestion 89:121, 1988
20. Itzkowitz
M, Okamura
on lectin
SH, Yuan
K, Umeyama
blood group Inst 79:425, 21.
Kim YS, Hakomoni
22.
Kobata
M, Ferrell
K, Jones
antigen
staining
Y, Hirota in human
expression
YS:
Histochemistry
RM, Chung
Cancer-associated
in the human
pancreas.
Itzkowitz SH, Yuan M, Chung YS, Satake 5: Le5 and Let antigen expression in human cer. Cancer Res 48:475, 1988 A, Ginsburg
V: Unidine
K, Umeyama pancreatic
Le Pendue, Oniol R,)uszczak tronJP: a-2-LFucosyltransferase
ficient 1983
red cells and normal
26. Lloyd KO: Blood group antigens and malignant change in human 27.
Mollicone
R, BaraJ,
G, Rouger
P, Salmon
34.
Schwyzwe
35.
Le PenduJ,
as markers for normal differentiation tissues. Am) Clin Pathol 87:129, 1987 Oniol
R: Immunohistologic
anti-
29.
Oniol
R, DanilovsJ,
Hawkins
BR: A new genetic
model
proposing
RS, Colcher
MA, Uchida D, Schlom
37.
‘Ji.kasaki H, Uchida E, Tempero MA, Burnett DA, Metzgar ES, Pour PM: Comparative studies on expression of CA 19-9 and DU-PAN-2 in pancreatic cancer tissue. Int ) Pancreatol 2:349, 1987
38.
Tempero MA, Uchida E, Takasaki H, Burnett DA, Steplewski Z, Pour PM: Relationship of carbohydrate antigen 19-9 and Lewis antigens in pancreatic cancer. Cancer Res 47:5501, 1987
39.
Uchida Presence
on their
that
group-specific N-acetylgalac) Biol Chem 252:2346,
properties.
E, BUchler M, Ness M), Burnett DA, ), Pour PM: Comparative studies on the expression of tumor-associated glycoprotein (Tag.2), CA 19-9 and DU-PAN-2 in normal, benign and malignant pancreatic tissue. Int) Cancer 42:681, 1988 Metzgar
53:219,
28. Mollicone R, DaIiX AM,)acobsson A, Samuelsson B, Gerard G, Crainic K, Caillard T, Le Pendu), Oniol R: Red cell.H-deficient, salivary ABH secretor phenotype of Reunion Island. Genetic control of the expression of H antigen in the skin. Glycoconjugate ) 5:499, 1988
A blood
II. Enzymatic
N: Blood group related Acta Pathol Microbiol
Szulman AE, Marcus DM: The histologic distribution ofthe blood group substances in man as disclosed by immunofluorescence. VI The Lea and Leb antigens during fetal development. Lab Invest 28:565, 1973
36. l#{224}kasaki H, Tempero
pattern
of type 1 (Lea, Le) and type 2 (X, Y, H) blood group-related gens in the human pylonic and duodenal mucosae. Lab Invest 1985
H, Dabelsteen E, Graem in human fetal pancreas.
M, Hill RI: Pocnine
tosaminyltransferase 1977
C, Car-
activity in sera ofindividuals with H-dcH antigen in secretions. Vox Sang 44:360,
Am)
Schenkel-Brunner H, Chester MA, Watkins WM: a-LFucosyltransferases in human serum from donors ofdiffenent ABO, secretor and Lewis bloodgroup phenotypes. Eur ) Biochem 30:269, 1972
diphosphate-N-acetyl-D-galactosamine:
G, Liberge
to the H gene.
33.
24. Lemieux RU, Baker DA, Weinstein WM, Switzen CM: Artificial antigens. Antibody preparations for the localization of Lewis determinants in tissues. Biochemistry 20:199, 1981 25.
linked
Pour PM, Tempero MM, Takasaki H, Uchida E, Takiyama Y, Burnett DA, Steplewski Z: Expression of blood group-related antigens ABH, Lewis A, Lewis B, Lewis X, Lewis Y, and CA-19-9 in pancreatic cancer cells in comparison with the patient’s blood group type. Cancer Res 48:5422, 1988
K, can-
23. Kumazaki T, Yoshida A: Biochemical evidence that secretor gene, Se, is a structural gene encoding a specific fucosyltransferase. Proc NatI Acad Sci USA 89:4193, 1984
closely
1981
32.
of
D-galactose a-3-N-acetyl-D-galactosaminyltransfenase, a product of the gene that determines blood type A in man.) Biol Chem 245:1484, 1970
gene
HIROTA
Philipsen EK, Clausen carbohydrate antigens Scand 96:1109, 1988
) NatI Cancer
1987
33:421,
OKAMURA,
31.
YS, Satake alterations
is a structural
NAKAJIMA,
30. #{216}rntoftiT, WolfH, Watkins WM: Activity ofthe human blood group ABO, Se, H, Le, and X gene-encoded glycosyltransferases in normal and malignant bladder unothelium. Cancer Res 48:4427, 1988
T: Effects of a-galactospancreas.
LD, Ratcliffe
RT, Kim
the Se gene Hum Genet
NISHI,
40.
Vierbuchen
E, Steplewski oftwo distinct
antigenicity.
Z, Mroczek E, Buehler acinar cell populations
Int ) Pancreatol
M, Uhlenbruck
G, Ortmann
1:213,
M, Burnett in human
D, Pour
pancreas
PM:
based
1986
M, Dufhues
G, Fisher
R: Oc-
currence and distribution of glycoconjugates in human tissues as detected by Erythrinacrtitagal/ilectin.J Histochem Cytochem 36:367, 1988 41.
Watkins WM: Genetics and biochemistry Proc R Soc Lund [B] 202:31, 1978
Downloaded from jhc.sagepub.com at VIRGINIA COMMONWEALTH UNIV on March 13, 2015
ofsome
human
blood
groups.