BIOLOGY
46, 328-334
REPRODUCTION
OF
(1992)
Human Endometrial Stromal Cells (CD) 13 Antigen/Aminopeptidase IUMITOSHI
Iivti,2’3
KARIYA,3
Department
MICHIYUKI
and
MAEDA,4
NOBUYUKI
EM!,3
KENJI
of Gynecology Kyoto
University,
and
Decidual Cells Express Cluster of Differentiation N and CD1O Antigen/Neutral Endopeptidase1
HIROSHI TAKAKURk,3
Obstetrics,3
54
Faculty Kawahara-cho,
FUJIWARA,3
NORIHIKO
HIDEHARU
KANZAKI,3
of Medicine,
and
OKAMOTO,3 and
Chest
Kyoto
Sakyo-ku,
MORI3
Research
Disease
606,
MASATOSHI
TAKAHIDE
Institute4
Japan
ABSTRACT With
specific
related
monoclonal
surface the
during
first
Indirect
trimester
of pregnancy
to aminopeptidase 93%
of
N and
the
enriched
examined
expressed
into
that
human
CD1O
activity
peptidase
and
was
on
detected
Human
endometrium
is composed
implantation ESC transform to play
of
an
origin
the fertilized into decidual
important
egg into cells (DC)
role(s)
of ESC
in the
and
DC is still
an other
increasing hemopoietic
dometrium
been
reported
whether ESC cells. By using
specific mouse and DC express
types
maintenance
in these
cell
we
and/or trimester
and
CD1O
antigen
the en-
were
Received
July
This
work
02857227,
supported
Japan and by the 1dorrespondence: rics,
Faculty
Shimizu Kimitoshi
of Medicine,
in part
by a Grant-in-Aid
the
Ministry
found that ESC (CD) 13 an-
shown
for
to be iden-
Scientific Science
Foundation Research Grant for 1990. lmai, M.D., Department of Gynecology Kvoto
University,
Sakyo.ku,
Kyoto
on
82cell-
stromal
on the
hydrolysis
606,
Research and
The was
peptidase activity also examined.
of
METHODS
prolapse deciduae
from
treatment
patients
who
of uterine
had
myoma
without hormonal therapy. were obtained from patients
First who
the date adjusted
ment
of the
of the last menstrual period and, by taking into account ultrasonic gestational
sac
and
fetal
if necessary, measure-
crown-rump
length.
1 shows
the
mouse
monoclonal
antibodies
used
tibody.
Indirect
frozen
(no.
Culture
obtained
for the
purchased from Becton Dickinson Immunocytometry Systems (Mountain View, CA). Fluorescein isothiocyanate (FITC)conjugated rabbit anti-mouse immunoglobulin (DAKOPAUS A/S, Glostrup, Denmark) was used as a second an-
and neutral enboth of which
of Education,
detected
legal abortions. Informed consent was obevery patient. Gestational age was calculated
from was
Immunofluorescence fresh
embedded
from
cells identical
in this study. My7 was purchased from Coulter Immunology (Hialeah, FL); MCS2, NU-Ni, NU-Ter, NTJ-B2, and Bear-i were purchased from Nichirei Co. (Tokyo, Japan), and LeuM3 was
1991.
03454396)
are
based
AND
were
hysterectomy
Table
with with
5, 1991.
was no,
Il,
which was
peptidases. suspension
cell
uterine human
The October
decidual
Antibodies
attempted
associated staining
3.4.11.2)[13] respectively,
function-
two
and
in endometrial
by an assay
endometria
had undergone tained from
stem is not
of studies on in the human
antibodies, we of differentiation
antigen,
3 antigen
cells
preparations
undergone
et al.
marrow that this
Here
CD1
examined
membrane-bound ESC-enriched
Human
of preg-
Kearns
number cells [8-121.
tical to aminopeptidase N (EC dopeptidase (EC 3.4.24.11)[14],
Accepted
analysis, the
CD1O
and
express cells
T&ues
through nutritional [11, [3] influences. reported on the surface of ESC and DC [4, 51, and
express antigens immunohistological
monoclonal both cluster
antigen
of
cells
stromal
MATERIALS
tigen and CD1O antigen. We also used flow cytometry to determine whether CD13 and CD1O antigens could be useful cell surface markers for ESC and DC in vitro. CD13
13 antigen
cytometric
75-93%
are the
the endometrium, and are considered
from bone evidence
the case. Recently, lymphocytes and to determine hemopoietic
of two
controversial.
[6] reported that DC are derived cells, but Fowlis et al. [7] provided
have
(CD)
decidual
and
endometrial
stromal cells (ESC) and endometrial The growth and differentiation of ESC of ovarian steroid hormones. After
nancy at the maternofetal interface endocrine [2], and immunological Only a few studies have been markers or functional parameters the
mainly
cells
both
alanine.
INTRODUCTION
of cells, endometrial glandular cells (EGC). are under the control
that
By flow
detected
was
stromal
revealed
of differentiation
respectively.
antigen
p-nitroaniline
endomethal
staining cluster
endopeptidase,
and
Furthermore,
of alanine-p-nitroanilide
found
immunofluorescence
neutral
cells,
preparations.
we
antibodies,
antigens.
tissue in OCT.
with
liquid
Staining
samples
were
compound nitrogen,
cut (Miles
and
kept
into Inc.,
small
pieces,
Elkhart,
at -80#{176}Cuntil
IN), sec-
tioning. Cryostat sections, about 6 tm thick, were air-dried and fixed with acetone at 20#{176}C for 5 mm. Nonspecific im-
of
-
and ObstetJapan.
munoglobulin 5% normal
FAX: 81-
75-761-3967.
328
binding was blocked rabbit serum in PBS (pH
by preincubation 7.4) for 15 mm.
with Excess
CD13 TABLE
1.
Monoclonal
antibodies
Antibody/CDa
used
MCS2/CD13 NU-N1/CD1O NU-B2/CD2O NU-Ter/CD2 Bear-1/CD11b LeuM3/CD14
as above neutral endopeptidase, Bgp135 sheep RBC receptor CR3, C3bi receptor, phosphoinositol-linked
serum
was
removed.
N
at an
first
antibody
appropriate
was
were Tokyo, replaced
on
Mac-i protein
ESC-enriched cell described elsewhere
suspension [23] with
cells
cALL,c
Dilution
granulocytes
natural keller macrophages
Subclass
1:40 1:40 1:40 1:10 1:40 1:10 1:10
cells
Reference
lgGl IgGi IgGi lgG2b IgGi IgGi lgG2b
15 15, 16 17 18 19 20, 21 21, 22
antigen.
The
sections
examined Japan). with
were
antibody three washes
then
FITC-conjugated another three For
incu-
in a moist with PBS, rabwashes
with a fluorescence negative controls,
normal
mouse
serum
dilution.
of ESC-Enriched
329
CELLS
leukemias
myeloid
the
endometria
with
scissors,
and
ring
in RPMI
1640
Cell Suspension was minor
prepared by a method modifications. Briefly,
FIG. 1. Indirect immunofluorescence staining of human as follows: a, MCS2/CD13; b, NU-Ni/CD1O; c, LeuM3/CD14; glands (G) are not (a,b). CD14 antigen-bearing cells, which dometrium Ic). The gland is elongated and slightly tortuous, Id). The black bar indicates 50 m. All pictures are enlarged
were
washed
incubated medium
times
in PBS,
at 37#{176}C with
three
continuous
(Flow
Laboratories,
minced
Irvine,
containing 1 mg/mI collagenase (Wako Pure Chemical dustries, LTD., Osaka, Japan), 0.005% deoxyribonuclease I (Sigma Chemical Company, St. Louis, MO), 100 U/mI icillin, 100 i.g/ml streptomycin (Flow Laboratories), and fetal calf serum (Flow RPM! 1640 medium, 1640
Preparation
STROMAL
hemopoietic
granulocytes,
as above lymphoid progenitor cells, B cells T cells granulocytes, monocytes, monocytes, granulocytes,
CALLAb
they were incubated for 30 mm with bit anti-mouse immunoglobulin. After sections (Nikon,
ENDOMETRIAL
reactivity
monocytes,
bated for 1 h with each monoclonal chamber at room temperature. After
the
HUMAN
in this study.
Cluster of differentiation antigen. bCommon acute lymphoblastic leukemia CCommon acute lymphoblastic leukemia.
the
ON
Main
aminopeptidase
with PBS, microscope
CD1O
Antigen
My7/CD13
rabbit
AND
medium,
and
Laboratories). the cells were ESC were
After three resuspended
separated
cell clumps by differential sedimentation bility of the cells was examined using exclusion test, and viability was always
from
stirUK) Intype pen10%
washes with in RPM! the
glandular
at 1 g. The viathe trypan blue dye greater than 95%.
endometrium in mid-proliferative phase using monoclonal antibodies. Tissues were stained d, hematoxylin and eosin. ESC are positively stained with CD13 and CD1O, but endometrial are considered to be tissue macrophages or granulocytes, are sparsely distributed in the enand the spindle-shaped ESC possess little cytoplasm, giving the appearance of naked nuclei equally.
330
IMAI
FIG. 2. Indirect stained as follows: endometrial glands gland is convoluted
Flow
Cytometric
A pellet ice with
immunofluorescence staining of human endometrium in secretory phase (cycle date 24) using monoclonal a, MCS2/CD13; b, NU-N1/CD1O; C, Bear-1/CD11b; d, hematoxylin and eosin. ESC are positively stained (G) are not (a,b). CD11b antigen-bearing cells are sparsely distributed Ic). The cytoplasm of ESC is enlarged (d). The black bar indicates 50 m. All pictures are enlarged equally.
of ESC-Enriched
Analysis
of 5 x
i05
cells
10 .tl of undiluted
was
incubated
primary
antibody
Preparation for
30 mm
listed
on
in Table
1. After two washes with Hanks’ solution containing 0.1% BSA (Sigma Chemical Company) and 0.1% NaN3, the cells were incubated for 30 mm with FITC-conjugated rabbit antimouse immunoglobulin on ice in the dark. The cells were then washed three times and suspended in Hanks’ solution to be analyzed by flow cytometry (FACscan, Becton Dickinson were
Immunocytometly stained with only
the first antibody was ulosa cell monoclonal published
results).
sity of the
endometria,
for each
Assay
prewarmed PBS (Merck, Darmstadt,
examined enzymatically of alanine-p-nitroanilide [24]. The ESC-enriched times with PBS and
containing Germany).
0.4
by a method into p-nicell suspenresuspended in
mM alanine-p-nitroanilide The incubation was carried
out in a water bath, with gende agitation, 37#{176}C. The reaction was stopped by addition
for 20 mm at of ice-cold 1.0
Japan).
Giken,
with The
cells den-
a specamount
by a standard
curve.
menstrual
cycles,
RESULTS
pausal
analyzed
examined
Tokyo,
formed was determined done in duplicate.
yr of age, of gestation,
were
4.2), and the The optical
nm was
of p-nitroanilmne All assays were
35-48 12 wk
cells
at 385
(Union
Endometria
secretory the basal ing
Staining
Immunofluorescence
in secretory and CD1O
and alanmne washed three
supernatant
replaced by mouse anti-porcine granantibodies (Fujiwara H. et al., un3000
antibodies. Tissues were with CD13 and CD1O, but and eosinophilic, and the
acid buffer (pH by centrifugation.
trophotometer
Indirect
At least
Peptidase activity was based on the hydrolysis troaniline sion was
M sodium acetate-acetic were rapidly removed
cells or
Systems). As negative controls, FITC-conjugated second antibody
sample. Enzyme
ET AL.
of 13 women and
7 were
phase. antigens
with
in proliferative
ESC, but not in proliferative
phase (Fig. 2a,b) layer also expressed
CDI1b
or
CD14
rophages
or
endometria hum and
(Figs. ic myometrium
granulocytes,
antigens (data not nancy, DC, which with round nuclei,
normal
deciduae from 11 women, were examined. Among the
of the menstrual both antigens.
antigens, were
phase,
EGC, expressed phase (Fig.
probably observed
and 2c). Endometrial were negative for
at 6 and premenoand
6 were
both CD13 la,b) and in cycle. ESC in Cells express-
monocytes/macsparsely
in the
surface epitheCD13 and CD1O
shown). In the first trimester of pregare identified as large, polygonal cells expressed both CD13 and CD1O anti-
CD13
AND
CD1O
ON
HUMAN
ENDOMETRIAL
STROMAL
FIG. 3. Indirect immnofluorescence staining of human decidua of 7 wk gestation. Tissues c, Bear-i /CD11b; d, hematoxylin and eosin. All pictures are enlarged equally. DC are positively antigen seems weak (a,b). The glands (G) are negative for CD13 antigens. Sparsely distributed the large and polygonal DC Id). The black bar indicates 50 p.m.
gens
(Fig.
3a,b).
the expression whereas that parison with secretory
Judging
from
the
fluorescence
intensity,
of the CDIO antigen on DC appeared weak of the CD13 antigen appeared strong in comantigen expression on ESC in proliferative or
phase.
The
glands
in the
decidua
expressed
ther CD13 nor CD1O antigens. The cells expressing or CD14 antigens were observed sparsely in the and they could be large DC (Fig. 3c). CD2 antigen-bearing
distinguished cells,
antigen-bearing
cells,
served
in both
sparsely
likely
from likely
the
and
CD11b and
Peptidase
were
immunoglobulmn,
Actimy
of the ESC-Enriched
amount
ofp-nitroanihmne
to the
number
of ESC
lation lated
also
ob-
0.994)
(data
not
coefficient: r to the incubation (data
not
Cytometric
The
Analysis
ESC-enriched
endometria secretory CD13
or
cell
women,
39-48
suspension
from
yr of age,
were
10
tact and
normally
analyzed.
Three
were in late proliferative phase, and 7 were in phase. No difference was observed between phases.
antigen and
fluorescence 4). CD1Ib, less than CD2
not
bind
antisame to
the
=
(from 0.998; time
Cell Suspension
formed 0.5
x
was 106
linearly
to 4 x
related
106;
corre-
Fig. 5. It was also linearly rebetween 5 and 60 mm (r =
DISCUSSION
menstruating
cells,
did
mouse to the
shown).
shown).
Flow
by the fact that which belong
of murine
The
CD2O
and
deciduae
was excluded cell antibodies,
nei-
polygonal
B lymphocytes,
were stained as follows: a, MCS2/CD13; b, NU-N1/CD1O; stained with C013 and CD1O, but the expression of CD1O CDilb antigen-bearing cells (c) can be distinguished from
to ESC granulosa
subclass ESC.
deciduae
T lymphocytes,
endometria
tibodies porcine
331
CELLS
CD1O
was
detected
antigen
was
on detected
82-93%
of the
on 75-93%.
examined Their
intensity seemed to be relatively strong CD14, CD2, or CD2O antigens were positive 10% of the examined cells (data not shown CD2O).
Nonspecific
binding
of CD13
and
CDIO
mean (Fig. on for an-
ESC and their direct descendants, DC, keep direct conwith semiallogeneic fetal antigens during pregnancy, are considered to play a very important role(s) in suc-
cessful pregnancy. There is no distinct functional parameter for ESC/DC, and only a few surface markers for ESC and DC have been reported [4, 5]. Therefore, with CD-series monoclonal antibodies, we attempted to determine whether ESC
and
DC
express
hemopoietic
cell-associated
and found that ESC and DC expressed nopeptidase N and CD1O antigen/neutral It is well established that lymphocytes, macrophages
are
distributed
in the
antigens
CD13 antigen/amiendopeptidase. granulocytes, and human
endometrium
332
IMAI
EF
AL.
400
a
FLI
FLI
FLI
U 400
400
TTT..,0
#{149}
0
ie FL
I
FL
Fluorescence FIG. 4. Flow cytometric analysis of stained as follows: a, a second antibody CD11b. Percent positivity was as follows:
Ie
I
o2 FL
io2
i#{248}
I
intensity
human ESC-enriched preparation from the endometrium in late-proliferative phase (cycle date 14). Cells were only as one of the negative controls; b, My7/CD13; c, MCS2/CD13; d, NU-N1/CD1O; e, LeuM3/CD14; f, Bear-li a, 2.1; b, 92.8; c, 93.1; d, 92.2; e, 3.9; f, 3.3. Over 90% of the cells were positive for CD13 and CD1O antigens.
C a) C C
(‘S 0 I-
z
& 1
2
No.
of
FIG. 5. Assay of peptidase activity of ESC-enriched (cycle date 23). The amount of p-nitroaniline hydrolyzed of ESC. Incubation time: 20 mm.
4
3
celis(x106) preparation from the from alanine-p-nitroanilide
endometrium is linearly
in mid-secretory related to the
phase number
CD13
and
decidua.
Using
specific
AND
monoclonal
that
ESC
and
DC
ON
antibodies,
be identified immunohistochemically tometry [12]. Our immunohistological clearly
CD1O
ENDOMETRIAL
HUMAN
they
can
18-111 or by flow cyfindings showed
express
both
CD13
and
CD1O
an-
tigens. Although CD13 and CD1O antigens are also expressed on granulocytes, monocytes and lymphoid cells [1517], these cells could be distinguished from ESC and DC with other specific CD11b, and CD14 ings on these works [8-12]. Judging
cell surface markers, such as CD2, antigens. Our immunohistological
hemopoietic from
the
cells
were
fluorescence
consistent
antigen
appeared that
to become
molecules expression
should
be
after
ascertained
by
CD13 antigen that of CD1O
a more
cytometric
analysis,
CD13
This
quantitative
antigen
Their
fluorescence that
intensity
CD13
and
was
CD1O
was
cells antigen
antigens
detected
in the ESC-enon more than
relatively
strong.
can
be
This
useful
cell surface markers for ESC. CD13 antigen has been shown to be identical nopeptidase N (EC 3.4.11.2) [13]. Ammnopeptidase integral, single-chain membrane glycoprotein that
of these
by ammnopeptidase ample, met-enkephalin,
der
intensively
membrane
on [25].
the
This
intestinal antigen
ulocytes, monocytes, myeloid [15, 16]. It is an ectoenzyme, on cell surface tive peptides,
and is also
kidney expressed
several
secretory
endometrium,
in
vitro
trimester dopeptidase
to amiN is an has been brush
on gran-
nm, tides
oxytocin,
enzyme activity.
1 (IL-i) [27]. and CD1O antigens are or not as ESC-enriched
activity in vitro, and found This does not necessarily
themselves
have
peptidase
activity
as enkephahins,
bradyki-
peptidases, we cell suspension
examhad
enzyme
The
authors
the flow
in the
female
genital
tract,
and
their
roles
has
day of the of human
secretion
[31]. In addition, to affect the
by
first
neutral immune
ensys-
successful
implantation
and
maintenance
may be useful for assessing en-
role(s)
of these
en-
wish
have
to thank
ytometric
Ms. Yumiko
been
Tomita
for
her
assistance
with
technical
analysis.
REFERENCES I. Krehbiel
RI-I. Cytochemical
pregnancy
of the decidual
studies
in the
and
production
reaction
of deciduomata.
in the
Physiol
early
rat during
Zool
1937;
10:212-
235. 2. Riddick AmJ
DH,
Kusmik
Obstet
TG,
human
PlC Suppression
Lala
Cell
S, Asano
M, Parhar
RS, Lala
monoclonal
of amniotic
S, Kano
1988;
fluid
prolactin.
J
on
Reprod
endometnial
Immunol
cell-specific
and
tissue
by early
alloreacuvity
116:392-410.
K. Studies
PK. Decidual
antobodies:
of lymphocyte
Immunol
antibodies.
by monoclonal
source
127:187-190.
decidua.
E., Mon
5. Kearns by
1977;
ItS, Kennedy
a possible
Decidua:
WF.
Gynecol
pregnancy
in human
10:1-14.
recognized J Immunol 1985;
surface
species
cells
1987; antigen(s)
distribution.
135:1046-1052. 6. Kearns,
M, Lala
dopregnant
PK. Bone
mouse
7. Powlis
DJ,
marrow.
Ansell
JD.
BR, Isaacson
11.
13.
Tabibzadeh Starkey
PM,
nancy
decidua: cytometty.
Look
Immunology
AT, Ashmun
glycoprotein
RA, Shapiro
CDI3
83:1299-1307. 14. Letarte M, Vera 1988;
cycle.
(gplSO)
5, Iran
leukemia
168:1247-1253.
of
in the
pseu.
from
bone
1988;
of leukocytic
Cell
evidence cells
in
Metab
populations
of large
isolation
of the
decidual
in early
J Pathol
lymphoid
sub-
127:66-73.
epithelium
Endocrinol
CWG.
and
human
that
1987;
leupreg-
endometrial
153:281-288.
human
endometrium
70:437-443. in human early preggranular lymphocytes by 1990;
65:129-134.
LI-I, Peiper
SC. Human
myeloid
to aminopeptidase.
is identical
R, AddisJBL, antigen
derived
characterization gland
J Clin
characterization
flow
1987;
lymphocytes.
IL, Redman
Sargent
not
distribution Pathol
A. Immunocytochemical activity
menstrual
are
cells
J
Am
are granulated
S. Proliferative the
lymphocytic
decidual
immunocvtochemical
D, Ritson
precursors
39:445-446.
endometrium.
granulocytes
cell
155:1537-1554.
PM. Immunohistological
throughout 12.
1982;
related to endometrial 1985; 55:35-44.
Hollings
stromal
of decidual
Med that
1985; PG. The
infiltrate Immunology
BulmerJN,
origin
J Exp
Evidence
in human
9. BulmerJN,Johnson
10.
marrow
uterus.
Transplantation
populations
ac-
tivity in vitro may be due to peptidase(s)-bearing cells, such as macrophages, but suggests that peptides could be degraded locally by the cells in the human endometrium. Some biologically active peptides are known to be present
expression
hCG
dometrial function. The physiological zymes remains to be clarified.
8. Kamat
pepand
that it had peptidase indicate that the ESC the
stimulates
trophoblasts been reported
to the
cocytic nancy.
because
mRNA
of pregnancy. Furthermore, these enzymes as surface markers and as new parameters
3. Parhar
so forth [17]. at the amino
substance P, neurotensin, chemotactic (fMet-Leu-Phe), gastrin, atrial natriuretic peptide,
interleukmn As CD13 ined whether
IL-113
in humans by degrading IL-i [32]. Taken together, ammnopeptidase N and neutral endopeptidase, located on the surface of ESC and DC, may modulate the metabolism of biologically active peptides and may
4. Nakayama
such
locally For exby both
tem
ac-
CD1O antigen has been shown to be identical to neutral endopeptidase (EC 3.4.21.11) [14], and is also an integral membrane protein, expressed on lymphoid progenitor cells, acute lymphocytic leukemia cells, subsets of B lymphocytes,
residues,
degraded
after the twenty-third inhibits decidualization
and
bor-
hin [26].
of hydrophobic
and
[30]
human has
gestational
side
be
ACKNOWLEDGMENTS
biologically and met-enkepha-
renal glomeruhi, and tubular epithehium, and It is also an ectoenzyme and leaves peptides
may
neutral endopeptidase. which can be degraded
in
leukemic cells, and so forth which exposes its catalytic site
[25], and hydrolyzes such as met-lys-bradykinmn
peptides
N or
vitro
studied
333
peptidases, is present in the uterine fluid of fertile women, and its role as an immunomodulator has been suggested [28]. IL-113, which can be degraded by neutral endopeptidase, has been shown to be present in the first trimester human deciduae more abundantly than in proliferative and
ESC
immu-
decidualization.
Some
CELLS
been reported to appear menstrual cycle [29]. IL-i
other
be differentiation-related this change in antigen
on more than 82% of the examined riched cell suspension, and CD1O indicates
the
studied.
contribute
method. By flow
75%.
of
of the whereas
weak
these antigens might of ESC/DC. However,
suggests
with
intensity
nohistological staining, the expression on ESC appeared to become strong
CD2O, find-
STROMAL
Onizuka
is identical
RJ, Quackenbush to
neutral
plasma N.J
Clin
membrane Invest
EJ. Common
endopeptidase
J. Exp
1989; acute Med
334 15.
IMAI
Gadd
S. Cluster
Schmidt Cell
In:
dem
Borne
Antigens.
Knapp
W,
AEGKr
Oxford,
New
DOrken (eds.),
B, Gilks Leucocyte
York, Tokyo:
WIt,
Rieber
Typing
Oxford
EP,
man
University
HG,
antibody
17. I)flrkcn
Sagawa
K, Menon
MCS-2 Jpn
J Cancer
In:
Knapp
routine
Res
13, N1fllkr’ P, Ik/JLtIIi) 0)10,
M, MinowadaJ.
in the
(Gann)
‘Pan-myeloid”
immunodiagnostic 1985;
Press;
IL 0llk.
the of
mono-
26.
leukemia
Fl’, Schmidt
0. H-Cell RE, SteIn
II,
von dent Antigens
Rome AB(ikr (eds, I, l,eucocyte Typing IV. WhIte Cell l)Iffrrcnllallon Oxford, New York, Tokyo: (.)xford llnlvct’sltv Press; 1989: 3%-34. 18. St.ishtttko F, Nadler l.M, I lards’ K, Schlossman SF. Charac’tertzath in f a human 13 lymphocyte specific antigen .1 Immunol I 980; 125:1678.I 685. 19. Kaim tun M, Marlin Fl. I lansen IA, Brown MA, Sladak AW, Nowluiski RC. Identl. ficallon of a human I lymphocyte surface Protein associated with the Frosette receptor. .1 F.xp Med 1981; I 53:207=212. 20. llreard I, Reinher-,. H., Kong PC, (,oldsteln 0, Schlossman SF. A monoc-lonal antibodv react lye with human peripheral blood mors vtes. 1 Immunol 1960, 12.1:194.5=
AJ, Maroux
peptidase
anti’
S. Topology
Physiol W, Zownir on
1981;
and
biologic
properties
of intact
biologically
Rev
of microvillar
1982;
cells.
J Im-
hydrolases
membrane
of kidney
62:91-128.
0, Behahl active
9.
Action
peptides:
of human kinin
pancreas
converting
alanine activity.
amino-
Clin
Chem
111:69-79.
EG, Skidgel RA. Neutral endopepildase 24.11 (enkephallnase) of peptkle hormones, FAsF.B j 1989; 3:145-151, 26. Petraglla F, Facchlnettl F, M’Futa K, Ruspa M, Mon&uvera JJ, Gandolfi AR. Endogenous oplolci poepticies in uterine fluid. FertIl SterIl 27.
nd
Erd04
related
regulators
F’, Genazzani 1966;
48:247=
251. 29.
30.
1948.
Todd Ill RE, Nadkr l.M, Schlossman SF. Antigens on human monocytes identified by tis ill, wIt mat ant hi idles. J Immunol 1981; 126:1 435= 14.12. 2.4. Osleen KG, Hill GA, llargrove,t1’, Gorstein F Development of a method to Isolate and culiure highly purilled populatli ins of stromal and eptihellal cells from hu. man endotnetrtal biopsy spetluflens. Fertil Steril 1969; 52985-972. 24. Anitiscaum, M, Alexander .1W, Babcock OF Surface .uminopeptldase ;ictiyl’ of ho.
1. Biochemical
142:1245-1252.
intestine.
Sidorowicz Acts
K, Mokienhauer
WK, Richer
1989;
25. Kenny
76:235-239.
A, Schwarz.Alhlcz
W, l)Orken
reagent: service
lymphocytes.
munol
IV. White
and
clonal
phenotvping.
21
AL.
782-784.
Drexler
gens:
CD13.
H, von
Differentiation
1989: 16.
report:
RE, Stein
ET
Kaum 5, Matt I), Strom 5, EIermti 1), Turner I. lnterleukln.l, antigen I IIA-DRo, and transforming growth factur.R expression placenta, and placental membranes Mn J Obstet Gynecol 1437 Karlya M, KnzakI II, Takakura K, lm’aI K, Okarnoto N, EmI N, lnterleukln.l inhibits In vitni deciduisllnsttots of human endometnial ClIn
il.
Endocrlnol
Metab
1991;
Oct
(In
human kukoe In endometrium, 1990;
163:1430-
Karlys
Y, Mon
stromal
T.
cells,
press).
Yagel 5, Lila PK, Powell WA, Casper RE. lnterleukln.l sttmulstes human rhoriolnic gotladotrtpln secretion by first trImester human trophoblast. J din En. docrinol Metab 1989; 68:992-995. 32. Plerart ME, Najdovskl I, Appelboom TB, Dcsrhmxlt.Lanckman MM. Effect of hu. man endopeptldase 24.11 (“enkephallna.se”) on IL-I -Induced thymocyte proliferation activity J Immunol 1988: 1403808-3811.