Mosaic differentiation of human villus enterocytes: Patchy expression of blood group A antigen in A nonsecretors

GASTROENTEROLOGY

1993;104:21-30

Mosaic Differentiation of Human Villus Enterocytes: Patchy Expression of Blood Group A Antigen in A Nonsecretors LUIGI MAIURI, VALERIA RAIA, ROBERTO FIOCCA, ENRICO SOLCIA, MATTE0 CORNAGGIA, OVE NOREN, HANS SJOSTROM, DALLAS SWALLOW, SALVATORE AURICCHIO, and

EPIK DABELSTEEN Department of Pediatrics, II Medical School, University of Naples, Italy; lstituto di Scienze dell’alimentazione, Consiglio Nazionale delle Richerche, Avellino, Italy; lnstituto di Ricerca e Cura a Carattere Scientifico, Policlinico San Matteo, Pavia, Italy; Multizonal Hospital, Varese. Italy; Department of Human Pathology, University of Pavia, Italy; Department of Biochemistry C, Panum Institute, University of Copenhagen, Denmark; Department of Oral Diagnosis, Royal Dental College, Copenhagen, Denmark; and Medical Research Council Human Biochemical Genetics Unit, the Galton Laboratory, University College London, England

Back#ound:The authors have shown that a mosaicism of brush border antigens may occur spontaneously on enterocytes of small intestine in human adult-type hypolactasia. The present paper gives another example of spontaneously occurring mosaicism as indicated by the patchy expression of blood group antigens on villus enterocytes. Methods: Thirty-five individuals were examined by lmmunomorphological techniques with antibodies against blood group antigens. Results: In 4 of 16 A blood group individuals, the blood group antigens were expressed only in some villus enterocytes. The individuals with this mosaic pattern were all shown to be nonsecretors. The A antigen in the positive enterocytes of these individuals was only present as the ALeb structure, whereas ALeY and ALed were also present in the secretors. The patches of positive enterocytes were randomly distributed along the villus wall. Condusions: A nonsecretor individuals express the bloqd group antlgens only in some villus enterocytes; this mosalcism does not arise from a heterogeneous population of stem cells within the crypts but rather reflects subtle differences in the pattern of differentiation between monoclonally derived epithelial cells on the villus.

of enterocytes the stem embryo and

aggregation

spontaneous

within

he small intestine is lined with a continuously regenerating and differentiating epithelium. Stud-

ies on rodents have shown that stem cells derived from a single progenitor and located in the crypts represent the source commences

of this epithelium. within the crypts

Cellular differentiation and proceeds during the

migration of the cells up the villus. Each crypt is composed of a single clone of cells.’ Several crypts contribute to each jejunal villus.2 It is well known that the pattern of differentiation of the enterocyte along the villus depends on the age of the cell and that it is equivalent in cells at a comparable location on the villus. It has been shown in animals that a mosaic pattern

on the villus

chimeras,‘T3 and

intestine.6

In X-linked

resulting

females’

villus,

e.g.,

transferase vertical

can be shown

deficiency

genotype

cellular

small

mosaicism in hetero-

histochemically guanine

on the

phosphoribosyl trans-

in humans.’

In all these cases

sheets or ribbons

of cells of a single

have been found

In contrast,

of the mouse

in mice’ and in ornithine

deficiency

continuous

mutations

inactivation

in hypoxantine

carbamylase

mice,4,5

somatic

disorders,

if

as it is in

transgenic

induced

from X-chromosome

zygous

surface

is heterogeneous,

the stem cell compartment

on the villus.

an “interrupted”

mosaic

pattern

of the

expression of some genes that are markers of enterocyte differentiation was recently described on the villi of rats during late gestation,” tern of staining was observed brane-associated proteins during the same period.” We have recently spontaneously indicated

T

may be induced

cell population

and

shown

brush

border

that mosaicism

on adult human

by the pattern

whereas a uniform patfor several other memenzymes also occurs

small intestinal

of lactase

protein

villi, as

expression

in some subjects with adult-type hypolactasia.12 In the present paper we give another example of spontaneously occurring mosaicism, as indicated by the variability in the expression of blood group A specificity of brush

border

components.

Materials

and Methods

Antibodies Polyclonal A antigen antibody 1686. This antiserum was generated inadvertently after immunization of a rabbit with purified human lactase from a few pooled individuals Abbreviations used in this paper: BSA, bovine serum albumin. 0 1993 by the American Gastroenterologlcal Association 0016-5085/93/$3.00

22

GASTROENTEROLOGY Vol. 104, No. 1

MAURI ET AL.

bodies

were used as undiluted

cept for the anti-Leb ascites

hybridoma

antibodies,

and used at a dilution

In addition antibodies

against

against

blood

group

carbohydrate

with

from

the

structures,

A a-1,3-N-acetylgalactos-

were used. This antibody

by immunization

ex-

were purified

of 1:400.

to antibodies

aminyltransferase

supernatants

which

purified

was established

native

A-transferase

protein.‘”

Light Microscopy Proximal from

small

informed

intestinal

Figure 1. Thin-layer chromatography.

lmmunostarning with rabbit antisera. Total neutral glycosphingolipids from blood group B, 0, or A (- 10 pg_/lane) and various purified glycoliprd standards (lanes 1-8, 0.5 @/lane) were applied at the origin. Plates, run in chloroformmethanol-water (50/40/10, vol/vol/vol) and blocked In 5% PBS-BSA, were subsequently incubated wtth rabbit serum (1:500 overnight), then with goat anti-rabbit lg for 1 hour, and finally with 125i-protein A for 1 hour. They were then dried and exposed by autoradiography. Lane 1, A type 1 chain; lane 2: ALeb type 1 chain; lane 3, A type 2 chain; lane 4, ALeYtype 2 chain; lane 5, A”(glob0 A) type 4 chain; lane 6, A type 3 chain; lane 7, AGMl type 4 chain; lane 8, AGAI type 4 chatn. Reactivity is observed with ALeb and ALeY in the 7-sugar region. Apparent binding to A” (lane 5) could be a result of contamination of the sample (see Results).

needed

duodenal

creatitis,

or gastric

A patchy

(two patients), before

according

second

group

week, four injections).13

fraction

20 c(g per injection

Uppsala,

The IgG fraction Western

blotting

biopsy

specimens [IO-mg

Elvehjem

1686 was analyzed

to lactase-phlorizin collected

individuals

specimens

small

with

different

homogenized

200,000

X g for 30 minutes

blood

cosphingolipid erythrocytes

group

activity

extracts

intestinal

Melsungen,

Germany),

standards’5,‘6

initial

studies,

monoclonal

against

group

antigens

A, B, and H purchased

Corp.

(Copenhagen,

Denmark)”

by Green

solution

histological

frozen

and kept at

procedures;

and kept at -80°C

sections

the

until used.

polyclonal

(working

for 20 hours

performed

as previously

oxidase body

with

with other culture

(1) phosphate specificity

hybridization).

were

using an immunoper-

replacement

Control

of the primary

(2) monoclonal

In addition, antiserum

and (3)

cell line SP 2 (used for

absorption

tests were performed

with Synsorb

Ltd., Edmonton,

anti-

antibody

but with the same Ig isotype, of the myeloma

on the polyclonal (Chembiomed

1:lOO) or monoclonal

immunofluorescence.*’ buffer,

on

were incubated

and the experiments

described12

included

supernatant

sections

dilution, at 4°C

or

technique2’ reactions

were cut and mounted

or frozen

antibodies

Surface The

(for

IgM and

antibodies from

from Ortho

mistry

Alberta,

A and Synsorb

B

Canada).

et al.‘s were used. Subsequently,

lmmunocytochemistry

also used. The details of these antibodies, their isotype, and the appropriate references are given in Table 1. All the anti-

surface

as described

was incubated

body (1:50 in PBS overnight directed

against

0.5% bovine peroxidase

a num-

ber of other monoclonal antibodies to Lea, Leb, and A with better defined and/or narrower (restricted) specificity were

three-dimensional

was performed

The sample

Diagnostic Systems (Milano, Italy) (antibody against blood group A antigen) and an IgG monoclonal anti-A (mbbml) prepared

drugs

One part of the

gly-

see legend to Figure 1 and Table 1). Monoclonal antibodies against blood group antigens blood

acetate

to routine

Five-micrometer

by thin-layer

using total neutral

glycolipid

or antibiotic

treatment.

Staining

from blood group A, B, or 0 human

as well as purified

gastric

at 4”C].

ABH. In the Dako

blood

in a Potter-

was shown

immunostaining’4

by

hydrolase

from

(B. Braun,

chromatographic

details

from

biopsy

A-Seph-

pan-

the four blood

saline (PBS) or was paraffin-em-

part was quickly

staining

homogenizer

centrifuged Anti-A

on protein

of the antibody

membranes

(1g)G

Sweden).

for binding

in microsomal

every

The immunoglobulin

was isolated by chromatography

arose (Pharmacia,

types

(about

of gastric

diagnosis:

cytostatic

surgical

was fixed in 4% form01

bedded

because

In particular,

did not receive

4°C in phosphate-buffered

with blood

taken

undergoing

gastric ulcer, and biliary tract stenosis.

slides. Paraffin-embedded of unknown

therapy

subjects had the following

(i.e., neomycin)

other

were

patients

ulcer, biliary tract diseases,

lymphoma.

cancer

The patients

surgical

or gastric

group

tissue

specimens adult

resection.

The patients cancer,

intestinal

and consenting

mouse

(Dako)

et al.”

with monoclonal

anti-A

at 4°C) followed

by rabbit

Ig (Dako)

serum albumin complex

immunohistoche-

by Schmidt

antiIg

[1:50 in PBS containing

(BSA)] and the peroxidase

anti-

(1: 100 in PBS-BSA).

Electron

Microscopy

Ultrathin

sections

from

paraformaldehyde-fixed,

Lowicryl (K4M)-embedded mucosal samplesz2 [Lowicryl (K4M)i Balzers Union, Siirstentiim, Liechtenstein] were incubated with the polyclonal antibody 1686 (working dilution,

I:100

overnight

at 4’C)

and then

with

gold-labeled

January

MOSAICISM

1993

Table 1.. Results

on a Blood

Group

Subjects

With

Mouse

Monoclonal

Antibodies

Used

in the

OF VILLUS

Present

ENTEROCYTES

Study Results

Mouse

Antigen Lea

Antigen determinant

Type chain 1

monoclonal

antibodies Antibody/ isotype

GalPl-3GlcNAcj3-R 4

References

on A subjects

With all enterocytes

With patchy distribution of A-positive

A-positive

enterocytes

Negative

BB and Golgi

BB and Golgi

Negative

37

BB and Golgi

BB and Golgi patchy

AH,$JgM

38

BB and Golgi

Golgi patchy

AH,,/JgM

38

BB and Golgi

Negative

W&G,

39

Negative

BB and Golgi

AH$lgM

40

Negative

Negative

HWG,

41

BB and Golgi

Negative

HHz/tgGa

37

BB and Golgi

Negative

HHJlgM

41

BB and Golgi

Golgi patchy

TH,/kW

42

BB and Golgi

Golgi patchy

IgG,

36

23

I Fuc 1 Leb

ALeb

1

1

GalPl-3GlcNAc81-R 2 4

I

I

Fuc 1 Fuc

1

Chembiomed, Edmonton, Canada

GalNAcal-3Ga@l-3GlcNAc81-R 2

A

1

HH$lgG,A

4

I

I

Fuc 1 Fuc

1

GalNAcal-3Galj31-R 2

I Fuc 1 ALed

1

GalNAcal-3Gal8

1-3GlcNAc8

1 -R

2

I

Fuc 1 LeX

2

GalPl-4GlcNAcbl-R 3

I Fuc 1 LeY

A monofucosylated

2

2

Gal8 1-4GlcNAc8 2 3

I

I

Fuc 1 Fuc

1

1 -R

GalNAcal-3Gal8

1-4GlcNAcR 2

I Fuc 1 ALeY

2

GalNAcal-3Galj31-4GlcNAc81-R 2

3

Fuc 1 Fuc

1

I

A

3

I

GalNAcal-3Gal8l-3GalNAc~l-O-Ser/lhr 2

I Fuc 1 A rep.

3

GalNAcal-3Gal8

1-3GalNAcal2

I

Fuc I BB, brush border.

24

MAIURI

GASTROENTEROLOGY

ET AL

Vol. 104.

No. 1

Light Microscopy Labeling by monoclonal antibodies against blood group antigens ABH. Jejunal specimens from 35 (16 group

subjects

A, 14 group

jects) were analyzed priate

blood

served

group

antigen;

on the brush

region

and was also evident cells (Figure

Figure 2. lmmunoreactlvlty with monoclonal antlbody against the A blood group antigens. lmmunoreactlvity In a blood group A subject with monoclonal antibody against blood group A antigen: the enterocytes of the VIM show intense lmmunoreactivity at the level of brush border and the Golgl apparatus. The endothelial cells of the blood vessels are also stained (paraffin-embedded tissue. Nomarskl optic. immunoperoxidase; origlnal magnification \450). A similar picture was observed in the 0 and 6 blood group subjects by lncubatlon with antibodies against appropriate blood group antigens (data not shown).

the samples

Ig polyclonal

antibodies

(I\’

San Mateo, CA) (1:50 for 1 hour at room trmpcraturc); were

then

ported.”

uranyl-lead

Control

incubations

distribution

thq

as previously

however, (Figure

Characterization 1686 was analyzed

tinal membranes

When

the anti-

by VC’estern blotting

of intes-

from people of blood group

A RhS,

a

strong reactivity was observed with several polypeptides of varying molecular weight but with no binding or very weak binding to the same glycoproteins from people of 0 Rh+ and B Rh+ blood groups (not shown). Blood group A specificity. The apparent blood group A specificity of the antibody 1686 was studied in detail using a comprehensive panel of different glycolipids carrying the A determinant. The anti-serum binds the difucosylated A type 1 and type 2 structures but not the monofucosylated structures (Figure 1). Some reactivity was observed with the globo-A structure

f.uc

an intensity

as a usef-ul

1686. Only intense

I

but it is conceivable that this activity was caused by a minor contamination with ALe”:‘AI.e’, because these components comigrate on thin-layer chromatography.

subjects

group

subjects

thelial

blood

was cells,

similar

to

12 individuals

the erythrocytes

intense

and

immunoreactivity

positive

control. 1686. The tisthe polyclonal

villus cells of A blood group

sub-

staining

and

in the brush (Figure

complete

border

4A-1). Blood group

absence

cross-reaction

of

was seen

(data not shown).

staining,

in B blood

No staining

\,essels cells or erythrocytes

in any of the subjects. A mosaic distribution

These

of staining

internal

region

showed

a weak

a patchy

reaction

of the cells.

cells showed

jects showed 0

blotting.

cells. The positive

%-I). In these specimens

the endothelial

whereas

by Western

showed

in the cells of the other

also in supranuclear

1686

antibodies

of inappropriate

.j subjects

one third

showed

that observed

antibody

re-

wet-c as for light microscopy.

of Antibody

individuals

group

of positive

in less than

Results Analysis body

counterstained

there was absence

monoclonal

Labeling by polyclonal antibody sue sections were also analyzed with

IAoratorirs,

was ob-

in the supranuclear

group.

and served goat anti-rabbit

from

B sub-

the appro-

in the red cells and endo-

of the

Four of 16 blood seen

and

2). In all subjects

of immunoreactivity blood

against

immunoreactivity

border

thelial against

0, and 5 group

with antibodies

of endo-

was observed

of immunoreactivity

at brush

border and Golgi level was also observed in the four blood group ,-\ subjects who had shown a patchy pattern with the monoclonal anti-:1 antibodies (Figure 3B). The staining

of the surface

epithelium

disappeared

after absorption of the polyclonal antibody by Synsorb A but not by Synsorb B in both A and B subjects (data not shown). In all the patchy subjects the sucrase-isomaltase protein was uniformly distributed in all the villus enterocytes (data not shown). Labeling by monoclonal antibodies with restricted specificity against type 1, type 2, and type 3 structures. The results are summarized in Table 1. All the antibodies to blood group A-related antigenic structures gave the same staining pattern as the polyclonal anti-A antibody in the 14 individuals with homogeneous staining pattern. tiowever, the only monoclonal anti-A antibody that gave a positive staining reaction in the mosaic subjects was the anti-Ale’. The pattern was similar to that seen with polyclonal antibody.

MOSAICISM OF VILLUS ENTEROCYTES

January 1993

25

Figure 3. Patchy reactivity in a blood group A subject. (A) lmmunoreactivity with monoclonal antibody against A blood group antigen. A patchy distribution of immunoreactivity is observed with the presence of some enterocytes with staining in both brush border and supranuclear region and some enterocytes showing absence of stainrng at any level. Note the presence of immunostaining on endothelial cells of the blood vessels (paraffin-embedded tissue, Nomarski optic, immunoperoxidase; original magnification X800). (8) lmmunoreactivity with polyclonal antibody 1686 in a subject showing a mosaic pattern with monoclonal anti-A antibodies: the immunoreactivity is present in some villous enterocytes at the level of the brush border and the supranuclear region; the other enterocytes show no staining at any level (paraffin-embedded tissue, Nomarski optic, immunoperoxidase; original magnification X 1600). (C) Patchy distribution of immunogold labeling in electron microscopic level with the polyclonal antibody 1686. Note the presence of gold particles on microvilli of one enterocyte while the two adjacent cells are negative. Arrowheads indicate cell junctions between adjacent cells (original magnification ~35,000). (D) lmmunogold labeling at an electron microscopic level with the polyclonal antibody 1686 on cisternae and vesicles of Golgi complex. Note gold labeling in a positive cell (original magnification X56,000).

All the enterocytes

of the 14 blood group A individ-

uals with the homogeneous staining pattern with antiA antibodies were negative with antibodies to Le” and Le” but positive with antibodies to Leb. In contrast all the enterocytes of the mosaic subjects were Le” and Le” positive and Leb negative. Labeling by antibodies to a- 1,3-N-acetylgalactosaminyltransferase. Antibodies against A geneencoded a - 1 ,3 - N- acetylgalactosaminyltransferase

stained the Golgi region of enterocytes of both groups of A blood group individuals in a similar manner (Figure 5). Surface immunocytochemistry. Samples from two blood group A subjects who had shown uniform staining at the light microscope level showed a uniform staining of the villus wall from the base to the top of the villi with the monoclonal anti-A antibody (Figure 64. In contrast, samples from one of the mosaic

26

MAIURI

ET AL.

GASTROENTEROLOGY

Vol. 104.

No. 1

Figure 4. lmmunoreactivity with polyclonal antibody 1686 in a blood group A subject. (A) The enterocytes of the villi show strong immunoreactivity of the brush border and milder staining of supranuclear region (paraffin-embedded tissue, Nomarski optic, immunoperoxidase; original magnification X 1400). (B) lmmunogold labeling with the polyclonal antibody 1686 at electron microscopic level over microvilli (original magnification X35,000) in a blood group A subject. (C) lmmunogold labeling with the polyclonal antibody 1686 at electron microscopic level in trans.Golgi cisternae (original magnification ~56,000) in a blood group A subject.

subjects

showed

were randomly

scattered

little

distributed

along

areas of staining the villus

that

wall (Fig-

ure 6B).

Electron Microscopy Labeling by polyclonal antibody 1686. Samples from two blood group A subjects who had shown uniform staining at the light microscope level also showed uniform labeling of the microvilli (Figure 4B) and of the trans-Golgi cisternae (Figure 4C). No staining was observed at all in two blood group 0 individuals (not shown). The intestine of one of the four blood group A individuals with mosaic pattern also showed a patchy distribution of immunoreactivity at the ultrastructural level. Some enterocytes showed reactivity over the microvilli (Figure 3C) and trans-Golgi cisternae (Figure 30); some enterocytes showed no immunoreactivity at all (Figure 3C). The ultrastructural features of both positive and negative cells were indistinguishable.

Discussion The ABH blood group antigens are the terminal carbohydrate structures found on certain glycoproteins and membrane glycolipids. They are found not only on the red cells but also on other epithelia such as the intestinal mucosa. Previous studies have shown the presence of A, B, and H blood group antigens in the surface epithelium and glands of the gastrointestinal tract.‘8~23,24 In the small intestine they occur in the goblet cells25 and also on the brush border of the absorptive cells.24 The blood group A determinant comprises the trisaccharide unit GalNAc

a-1-3

Gal /31R

Fuc i that is carried on different core structures. Differences in the carrier core structure, but not in the A determi-

January 1993

MOSAICISM OF VILLUS ENTEROCYTES

27

type 2H, Gal 1-4 GlcNAc-R) 2

(type lH, Gal 1 3GlcNAc-R; 2

Fuc 1

Fuc 1

and acts preferentially

on type 1 chains.

do not have this enzyme.

Previous

cated that the expression

of the ABH

Nonsecretors

studies

have indi-

antigens

in the

small intestine is largely under the control of the secretor gene INCUS,‘* although some cells in the deeper part of the crypts and in the Brunner’s num express the antigens see reference present four

observation

study of patches

of stained

example

A antigens

of the duode-

in nonsecretorsz4

18). The

individuals

ported

glands

represents

(for review

described

another

previously

of secretor-independent

in the small

intestine.

in the

cells on the villi of unre-

expression

The mosaic

of

pattern

of expression is reminiscent of the staining of the Brunner’s glandsz4 with similar reagents, but in that case the mosaic pattern is seen in both secretors and nonsecretors. The most likely explanation for the presence of ALeb antigen tered activity however, Figure 5. lmmunoreactivlty

with antibody anti a- 1,3-N-acetylgalactosaminyltransferase in a subject with patchy distribution of immunoreactivity of A antigens. Note the staining of the Golgi apparatus in all enterocytes (cryostat sections, immunofluorescence).

kind,

in these cells is an increased

it could

also reflect

such as differences

variation

nant

itself,

provide

the basis for the presence

and immunologically

distinct

A antigenz6

(Figure

1).

Four of 16 blood expression lium,

6 and Table group

of the A antigen

whereas

A individuals

of the of the

had a patchy

on the intestinal

the A gene-encoded

lactosaminyltransferase

forms

epithe-

a- 1,3-N-acetylga-

was uniformly

present

in all

of the

Lewis

the cells. The

expression

structures,

similar

on

the

enterocytes

to that seen in salivary

glands

from

nonsecretors,” suggests that the four subjects with mosaic pattern who express Le” and Le” but not Leb antigens in the villus enterocytes whereas the other A individuals

are the nonsecretors that express Leb but

not Le” and Le” are the secretors.

The

secretor

gene

encodes one of the two major a-1,2-fucosyltransferases responsible for building the H structure that is the precursor to A

al-

in amount

of some other

of one of the sub-

strates, e.g., type 1 core structure. Whatever the mechanism, the results show an underlying variability of the differentiation pattern of enterocytes along the villus. It is clear that not all the genes expressed

structurally

and/or

of one of the two a-2-fucosyltransferases;

in the fully

differentiated cells are affected, because we have shown the sucrase-isomaltase staining to be uniform. The mosaic pattern is probably not caused by cell cycle-dependent effects, because cellular known to be confined to the crypts

proliferation (for review

is see

reference 28); it also appears not to reflect differences in cellular architecture, because the cells could not be discriminated by electron microscopy. The mosaic pattern of enterocytes along the villus could reflect the presence of two (or more) populations of enterocytes, each of which has arisen from a different stem cell. However, the staining of A blood group substances on the surface of the villus has shown that positive cells in nonsecretors are randomly distributed on the villus and that there are no ribbons or sheets of positive cells arising from the crypts, as one would expect if the origin of this phenomenon were clonal. We therefore conclude that most probably the mosaic pattern reflects the presence of subtle differences of the pattern of differentiation between monoclonally derived epithelial cells on the villus.

28

MAIURI

GASTROENTEROLOGY

ET AL.

Vol. 104,

No. 1

A Figure 6. Surface immunoreactivity in blood group A subjects. (A) Surface immunocytochemical staining with the anti-A monoclonal antibody in a subject showing a uniform pattern on tissue sections with monoclonal and polyclonal anti-A antibodies. Note the staining of all the enterocytes on the villus wall. The unstained areas correspond to the goblet cells. Note also intensely stained enterocytes in the crypts (immunoperoxidase; original magnification x60). (6) Surface immunocytochemical stainingwith the anti-A monoclonal antibody in a subject showinga patchy pattern on tissue sections with monoclonal and polyclonal anti-A antibodies. Note some intensely stained enterocytes randomly distributed along the villus wall, surrounded by a majority of negative areas. Note also the strong staining in some cells in the deeper part of the crypts (immunoperoxidase; original magnification X60).

CHAIN PERIPHERAL CORE TYPE 1

TYPE 2 Galpl-4GlcNA+-R

Gal/U-3GlcNAcal-R

e-u-R

e-0-R

??

LEA

R

TYPE

1


enzyme

C-JR

-F

@+R

CHAIN

<

X-enzyme

SIR H(k)

enzyme

enzyme I

1

-__BR

< Lew's

eWme

??

A

LEX

Se(H)

??

LEE

TYPE 2

CHAIN

LEY

H

WaR

SaR

<

A

A A

X-enzyme

0

icaR

H A

enzyme

ALan

ALEX

;

•~a1

H

:

??

G~cNAc

;

A

(monafucosyl

FUC 1-2

;

)

?? FUC

enzyme J

I

A( monofucosyl)

ALEX

l-4

;

OFUC

l-3

;

HGalNk.

H substance

Figure 7. Biosynthesis of A and Lewis antigens on type 1 and type 2 chain poly-N-acetylactosamine structure. A and Lewis antigens may be carried by type 1 and type 2 core structures, both of the lacto-type (j%galactosyl-N-acetylglucose). Two distinct a- 1-2-fucosyltransferase, encoded by the secretor and H genes, respectively, are presently believed to exist. 31-33The secretor (Se) gene-encoded fucosyltransferase has preference for type 1 chain substrate, whereas the H gene-encoded transferase has preference for the type 2 chain.34 The Lewis gene-encoded a- I-4-fucosyltransferasemay use both the type 1 and the type 2 chain substrates, thus participating in the formation of both LeaD and Lex’y.35 Independent a- I-3-fucosyltransferaseactivities have been recognized and are believed to be encoded by the X gene.35 The A gene-encoded a- 1-3N-acetylgalactosaminyltransferase may use both type 1 (Led) and type 2 (H) chain substrates.

January 1993

Heterogeneous expression of cell-surface antigens has been shown in normal epithelia and in their tumors, as shown by monoclonal and polyclonal antibodies.29 The functional implications of the present observation are not clear; it is known that oligosaccharides play a vital role in a variety of surface-related functions, including cell differentiation, recognition, and cell adhesion. Heterogeneity of the surface carbohydrates of enterocytes may imply variations in the digestive, transport, and receptor functions of the intestine. As a possible related model, a cell culture system has been described in which cell variants lacking the blood group A antigen display an epidermal growth factor receptor with increased affinity for its ligand.30

References 1. Ponder BAJ, Schmidt GH, Wilkinson MM, Wood MJ, Monk M,

2.

3.

4.

5.

6.

7. 8.

9.

10.

11.

12.

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Received June 11, 1991. Accepted August 4, 1992. Address requests for reprints to: Prof. Salvatore Auricchio, Dipartimento dl Pediatrla, II Facolta’ di Medicina, Universita’ di Napoli, Naples, Italy. The authors thank Dr. H. Clausen, Department of Oral Pathology, Royal Dental College, Copenhagen, Denmark; Dr. Andrea Quaroni, Department of Physiology, Cornell Unlversity, Ithaca, New York, for the gift of the anti-sucrase and anti-isomaltase monoclonal antlbodies; and Dr. Francesco Paparo, Department of Pediatrics, II Medical School, University of Naples, Italy, for technical assistance. This work has been presented in part at the 3th ESPGAN-NASPG joint meeting in Amsterdam, May 1990.