(CANCER RESEARCH 51, 1300-1307, February 15, 1991|
Expression of the Natural Killer Cell-associated Antigens CD56 and CD57 in
Human Neural and Striated Muscle Cells and in Their Tumors1
Gunhild Mechtersheimer, Martina Staudter, and Peter Möller2
Institute of Pathology; University of Heidelberg, Heidelberg, Federal Republic of Germany
ABSTRACT
The expression of the natural killer cell-associated antigens CD56 and
(1)57 which are known to show sequence homologies to the neural cell
adhesion molecule N-CAM was examined immunohistochemically
in
normal and regenerative human neural and striated muscle cells and in
their tumors using monoclonal antibodies Leu-19 and Leu-7. The pattern
of expression of CD56 and CD57 in neural tissue was assessed in
comparison with that of M, 68,000 neurofilament. Dendritic interstitial
cells were discriminated from neural cells by application of the pan
leukocyte antigen CD53.
In normal tissue, CD56 was expressed in thin nerve fibers, fine varicose
and sensory nerve endings, cell membranes of ganglion cells, and fetal
striated muscle cells. Thick nerve fibers, perikarya of ganglion cells, and
adult striated muscle fibers were CD56 negative. In the normal state,
CD57 was restricted to thick nerve fibers. Enhanced expression or
reexpression of CD56 was found in regenerative neural cells which, in
part, were also CD57 positive and in regenerative, CD57-negative striated
muscle cells. In neural tumors, CD56 was detectable in 3 of 3 benign and
8 of 13 malignant schwannomas, 1 of 4 peripheral neuroepitheliomas, 4
of 4 ganglioneuromas,
and 8 of 8 (ganglio-)neuroblastomas,
whereas
CD57 was restricted to one benign and one malignant schwannoma.
Furthermore, CD56, but not CD57, could be found in 8 of 8 rhabdomyosarcomas. All in all, the pattern of expression of CD56 is comparable to
that of N-CAM. CD57 exhibits a very restricted binding pattern which,
in most instances, is complementary to that of CD56. The absence of
CD56 in some neural tumors might reflect alterations in cell-cell inter
actions. Its reexpression in regenerative striated muscle cells and in
rhabdomyosarcomas suggests a role of CD56 as an oncodevelopmental
antigen.
INTRODUCTION
CD56 and CD57 are newly defined clusters of leukocyte
differentiation (CD)3 antigens corresponding to the NK3 cellassociated molecules NKH-1 and HNK-1, respectively.
CD57(HNK-1) comprises three antibodies one of which is Leu7, raised against the human T-cell line HSB-2 (1). In hematopoietic tissue, CD57 is present in 15-20% of peripheral blood
mononuclear cells, including 60% of NK-active cells, and a
subset of peripheral blood cells without NK activity (2, 3).
CD57 detects an A/r 110,000 glycoprotein. However, since the
antibody-binding site is a carbohydrate structure, there might
be more than one molecular structure accounting for apparent
differences in published molecular weights. Furthermore, CD57
has been found to recognize the myelin-associated antigen (4)
and it also detects a sulfated carbohydrate epitope expressed on
a portion of N-CAM isoforms (5).
CD56 consists of six antibodies including Leu-19. The Leu19 antigen is an M, 220,000/140,000 cell surface glycoprotein
(6-8) corresponding to the NKH-1 molecule (9, 10). It is
expressed on virtually all human natural killer cells and on a
subset of T-lymphocytes and interleukin-2-activated
thymocytes that mediate major histocompatibility complex-unre
stricted cytotoxicity (6, 9, 10). Additionally, it has been found
on certain CD4+ thymocyte cell clones, some myeloid leukemias, and the KG la immature hematopoietic leukemia cell line
(7). Recently, Lanier et al. (8) found that CD56 was expressed
also in neural tissue and they were able to demonstrate its
identity to the A/r 140,000 isoform of the neural cell adhesion
molecule N-CAM.
N-CAM, an integral membrane glycoprotein, is one of the
best studied cell adhesion molecules (11). It is a member of the
immunoglobulin supergene family (12) and mediates Ca2+independent homophilic intercellular binding (11). A consid
erable number of molecular isoforms have been found which
make up two main categories, namely, brain and muscle NCAM (12-14). Furthermore, soluble forms of N-CAM have
been isolated (15, 16). It is now known that these major iso
forms are generated from a single-copy gene located on band
q23 of human chromosome 11 (17) by alternative RNA proc
essing at the splicing and polyadenylation levels (12, 18). They
differ mainly in the amounts of sialic acid at the carboxyterminal ends of the molecules, reflecting different stages during
embryogenesis (19).
In the normal state, N-CAM is present in all three germ
layers during early development (20). In the adult, however, it
is mainly restricted to neural cells (11). Thus, N-CAM has been
found to play a central role in various developmental events
including the orderly outgrowth of axons (21), cell pattern
formation (19), and nerve-muscle interactions (22). Further
more, it is assumed that cell-cell interactions mediated by NCAM act as regenerative key mechanisms since N-CAM is
present in regenerative neural and striated muscle tissues (23,
24).
Although the distribution pattern of N-CAM has been exten
sively studied in vertebrates, information concerning its pattern
of expression in human tissue is still scarce. The identity of
CD56 and CD57 molecules with the M, 140,000 isoform and
with at least a portion of N-CAM, respectively, prompted us to
study their expression in a comprehensive series of normal
neural and striated muscle cells and in related tumors by appli
cation of mAbs Leu-19 and Leu-7, respectively.
MATERIALS AND METHODS
Received 8/3/90; accepted 11/20/90.
The costs of publication of this article were defrayed in part by the payment
of page charges. This article must therefore be hereby marked advertisement in
accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
1This study was supported by a grant from the Deutsche Krebshilfe/Dr.
Mildred Scheel-Stiftung für
Krebsforschung (W50/89/MÖ 2).
2To whom requests for reprints should be addressed, at Pathologisches Institut
der Universität Heidelberg, Im Neuenheimer Feld 220, D-6900 Heidelberg,
Federal Republic of Germany.
'The abbreviations used are: CD. cluster of designation; NK, natural killer;
mAb, monoclonal antibody; AEC, 3-amino-9-ethylcarbazole: NF. neurofilament:
D1C, dendritic interstitial cells; RMS, rhabdomyosarcoma.
Tissue. Most tissues, including all tumors, were obtained from sur
gical specimens. Some nonneoplastic tissues were drawn from autopsy
material during the first 6 h after death. Diagnosis of the tumors was
based on standard histopathological criteria as described by Enzinger
and Weiss (25). The tissue samples were frozen quickly in liquid
nitrogen and stored at —¿70°C.
Serial frozen sections of about 1 cm2
and a thickness of 4-6 urn were dried in air, fixed in acetone at room
temperature for 10 min, and immediately stained or stored at —¿
20°C
for a short period.
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CD56 AND CD57 IN HUMAN NEURAL AND STRIATED MUSCLE TISSUES
Reagents. mAb Leu-19 (isotype IgGl) recognizing the CD56(NKH1) antigen and mAb Leu-7 (isotype IgM) directed against the
CD57(HNK-1) antigen were obtained from Becton-Dickinson (Moun
tain View, CA); mAb NR4 (isotype IgGl) directed against M, 68,000
neurofilament was purchased from Dakopatts (Copenhagen, Denmark).
MAb HD77 [isotype IgM(Gl)] recognizing a recently defined pan
leukocyte antigen designated CD53 (26) was raised in our own labora
tory (in collaboration with Dr. B. Dörkenand Dr. G. Moldenhauer).
CD22 mAb HD39 (isotype IgGl) and CD21 mAb B2 (isotype IgM).
which were used as isotype-matched control antibodies, were provided
by B. Dörkenand G. Moldenhauer (Heidelberg, Federal Republic of
Germany) and by L. N. Nadler (Boston, MA), respectively. A polyclonal
biotinylated sheep antibody to mouse immunoglobulin (reactive with
all mouse isotypes) and a streptavidin-biotinylated peroxidase complex
were provided by Amersham (High Wycombe, United Kingdom). AEC
and N'N-dimethylformamide were obtained from Sigma Chemical Co.
(St. Louis, MO).
Staining Procedure. After rehydration with phosphate-buffered saline
solution (pH 7.5), the frozen sections were incubated for l h with
purified mAbs at appropriate dilutions (Leu-19, 1:20; Leu-7, 1:20;
NR4, 1:50; HD77, 1:100). The sections were then incubated with
biotinylated anti-mouse immunoglobulin (1:50) and streptavidin-bioti
nylated peroxidase complex (1:100) for 30 min, respectively. All incu
bation steps were carried out in a humid chamber at room temperature
and then rinsed twice in phosphate-buffered saline. The peroxidase
reaction product was reddish brown when AEC and H2Oz were used as
substrate: 4 mg AEC was dissolved in 500 n\ yVW-dimethylformamide.
Subsequently, 9.5 ml sodium acetate buffer (0.05 M; pH 5.0) and, just
before use, 5 n\ H2Û2(30%) were added. The incubation time was 15
min. The sections were then rinsed in distilled water, counterstained
with Harris' hematoxylin, and mounted in glycerol gelatin.
Controls. Negative controls in each case were performed without the
primary antibody; no staining was observed except for scattered granulocytes. This staining was due to endogenous peroxidase, which was
not blocked for the benefit of optimal antigenicity. The positive results
were confirmed by controls using the isotype-matched mAbs HD39
(1:100) and B2 (1:100), which yielded negative results in all cases,
ruling out nonspecific antibody binding. Minor peripheral nerve trunks
present in almost all tissue samples studied served as positive intrinsic
controls for the reactivity of CD56, CD57, and NF mAbs, respectively.
Furthermore, expression of CD56 and, to a lesser extend, of CD57 in
scattered CD53-positive lymphoid cells proved the immune reaction to
be reliable.
Evaluation. Detailed expression of CD56 and CD57 antigens in
peripheral nerves, sensory nerve endings, autonomie ganglia, and neural
tumors was assessed in comparison with the reactivity of NF in these
tissues. mAb CD53(HD77) was used, and the visualization of DIC, in
addition to lymphoid cells, which were abundant especially in neural
tissue, allowed a clear discrimination between nonneoplastic and neoplastic neural cells. In neoplasias, the staining of tumor cells was
evaluated in a semiquantitative fashion: +, entire tumor cell population
positive; ±,positive and negative tumor cells in various amounts (for
details see "Results"); -, entire tumor cell population negative.
RESULTS
The expression of CD56 and CD57 antigens in nonneoplastic
neural and striated muscle cells as well as in their tumors is
shown in Tables 1 and 2.
Normal Neural Tissues. The analysis of normal peripheral
nerve trunks showed expression of CD56 in thin CD57~ nerve
fibers. Thick CD57+ nerve fibers lacked any detectable CD56
molecules (Fig. 1, A and B). Both thin and thick nerve fibers
were NF+ (Fig. 1C). Staining with mAb CD53 showed a con
siderable number of DIC within the major nerve trunks and
thus allowed clear discrimination between neural cells and DIC
(Fig. ID). Minor peripheral nerve trunks present in various
tissues exhibited the same antigenic profile as major peripheral
nerve trunks. In addition, fine varicose nerve fibers surrounding
Table 1 Expression ofCD56 and CD57 antigens in nonneoplastic neural and
striated muscle cells
Cell type
Thin peripheral nerve fibers
CD56
+"
Thick peripheral nerve fibers
Fine varicose nerve fibers
Regenerative nerve fibers
Meissner's corpuscles
+
+
+
Pacinian corpuscles (axons)
+
CD57
+
+/-
Pacinian corpuscles (lamellae)
Intrafusal muscle fibers
+/—
Ganglion cells
+
—¿
Satellite cells
+
Chromaffine cells (adrenal medulla)
+
+/(+)
Striated muscle fibers (first trimenon)
+
Striated muscle fibers (third trimenon)
(+)/Striated muscle fibers (adult)
-/(+)*
Striated muscle fibers (regenerative)
+/(+)
"Scoring of cell reaction: +, strong staining intensity for all cells; (+), weak
staining intensity for all cells; •¿. positive and negative cells in various amounts;
-, negativity for all cells.
* Weak positivity in some muscle fibers of eye muscles and of the tongue.
Table 2 Expression ofCD56 and CD57 in tumors of neural and striated
muscle origin
CD56/CD57
PhenotypeBenign
+°3
3/-13
schwannomaMalignant
5/-4
schwannomaPeripheral
_/_44/—4/-4-/44/—4
neuroepitheliomaGanglioneuromaGanglion
cellsSchwann
cellsGanglioneuroblastomaGanglion
cellsNeuroblastsNeuroblastomaRhabdomyosarcomaN
/4-/I—
/34/——
/4-/4—
/——
4/-8
/-2/--~/25/123/4—
/4-/8
6/-+/--/I3/1l/--/-—
°Scoring of tumor cell reaction: +, strong positivity for all tumor cells; +/strongly and/or weakly positive and negative tumor cells in various amounts; negativity for all tumor cells.
cutaneous appendages and the base of crypt cells in a bowel
tissue specimen expressed CD56 antigens in the absence of
CD57 and NF molecules (Fig. JA). Meissner's corpuscles and
axons of Pacinian corpuscles were CD56+ but CD57~ (Fig. 2,
B and C); they also expressed NF. Surrounding lamellae com
posed of flattened spindle cells were CD56~/CD57~/NF~. The
perikarya of ganglion cells of peripheral ganglia were strongly
NF+ but lacked any detectable CD56 and CD57 molecules; the
satellite cells were CD56+ but CD57~ and NF~ (Fig. 3). Scat
tered DIC surrounding the ganglia were discriminated from
CD56* satellite cells by expression of CD53 antigen. Expres
sion of CD56 on the surface of ganglion cells was difficult to
assess because of the strong CD56 positivity of surrounding
satellite cells. Yet, in ganglioneuromas and optionally also in
ganglioneuroblastomas, immature ganglion cells lacking satel
lite cells clearly showed membrane-bound staining for CD56
molecules (cf. below). Chromaffine cells of the adrenal medulla
were CD56+CD57+/(+)/NF+. Altogether, CD56 and CD57 dis
played in most instances a complementary pattern of expression
with an overall predominance of CD56.
Regenerative Neural Cells. In traumatic neuromas the over
whelming majority of haphazardly oriented nerve fibers were
CD56* (Fig. 4A). In addition, and in contrast to the normal
state, serial section preparations disclosed expression of CD56
antigen also in at least some of the CD57* nerve fibers (Fig.
4B). NF was consistently expressed in traumatic neuromas.
Tumors of the Peripheral Nervous System. In three benign
schwannomas, the entire spindle tumor cell population was
CD56+. CD57 and NF, however, were restricted to a tumor cell
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CD56 AND CD57 IN HUMAN NEURAL AND STRIATED MUSCLE TISSUES
subpopulation of one case, respectively. Five of 13 malignant
schwannomas exhibited strong and consistent expression of
CD56 in the entire tumor cell population (Fig. 5/1); in 3 of 13
tumors at least one-half of the neoplastic cells were CD56+
(Figs. 5B and 9Ä),whereas 5 of 13 cases lacked any detectable
CD56 molecules. CD57 and NF were each restricted to one
case within a minor tumor cell subpopulation. Three peripheral
neuroepitheliomas were CD56~ throughout (Fig. 8/1); in one
case some scattered undifferentiated tumor cells expressed
CD56 antigen. CD57, like NF, was absent in all peripheral
neuroepitheliomas studied. Thin CD56+ and thick CD57* nerve
fibers, which were NF+, and/or scattered CD56+/CD57+
. 'p '¡.a*
y
''•y
lymphoid cells served as positive intrinsic controls for the
immune reaction in CD56" and/or CD57~ tumors, ruling out
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false-negative staining (Fig. 5C).
Tumors of Autonomie Ganglia. Some of the ganglion cells of
four ganglioneuromas studied lacked satellite cells and showed
clearly discernible CD56+ cell membranes; mature ganglion
cells in addition showed CD56+/CD57~/NF~ satellite cells. The
<
majority of Schwann cells were CD56+, whereas CD57 was
only present in about one-half of the Schwann cell compartment
in one ganglioneuroma and in two cases within a minor
Schwann cell population; at least some of the CD57+ Schwann
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cells coexpressed CD56. NF was consistently expressed in the
Schwann cells of ganglioneuromas. Immature ganglion cells
present in four ganglioneuroblastomas lacked satellite cells and
showed CD56'/CD57~/NF+
perikarya; some but not all NF+
'
ganglion cells exhibited membrane-bound staining for CD56,
as did all CD57~/NF" small, round, undifferentiated neuroblasts in these cases as well as in four undifferentiated neuroblastomas (Fig. 6). Additionally, extracellular CD56+ deposits
t
'.*
*>
-S*
were found around some larger neuroblasts (Fig. 6/1).
Normal Striated Muscle Cells. Fetal skeletal muscle cells of
18 weeks of gestation were strongly CD56+ and those of 35
weeks were weakly CD56+ compared to strongly CD56+ thin
nerve fibers of adjacent minor peripheral nerve trunks (Fig.
1A). Adult skeletal muscle cells were CD56~ and CD57~
Fig. 1. Serial section preparation of a major peripheral nene trunk (peroneal
nene). In A. CD56 detected by mAb Leu-19 is confined to thin nerve fibers. In
B, mAb Leu-7, in contrast, shows expression of CD57 in the myelin sheath of
thick nene fibers. In C, a neurofilament visualized by mAb NR4 is expressed in
both axons of thick and thin nerve fibers. In D, CD53 illustrated by mAb HD77
allows a clear discrimination between nene fibers and dendritic interstitial cells.
Immunoperoxidase staining of frozen sections, faint hematoxylin counterstain;
same technique for all photographs; x 145.
throughout (Fig. IB and C). A minority of muscle fibers of the
tongue and of eye muscles which were extremely rich in minor
peripheral nerve bundles and fine varicose nerve fibers were
CD56+ in the absence of CD57. In muscle spindles included in
an upper extremity skeletal muscle, at least some of the intrafusal muscle fibers were CD56+ and lacked CD57 molecules
(Fig. IB and C).
2C
2B
Fig. 2. Skin specimen of the finger tip. CD56 is strongly expressed in A, a dense network of fine varicose nerve fibers surrounding cutaneous appendages and blood
vessels (x 145). B, Meissner's corpuscles found in dermal papillae (x 145): C, axons of Pacinian corpuscles while absent in surrounding flattened lamellae (x 90).
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CD56 AND CD57 IN HUMAN NEURAL AND STRIATED MUSCLE TISSUES
* .
se
Fig. 3. Serial section preparation of a sympathetic ganglion (thorax). In A, the perikarya of ganglion cells and adjacent nerve fibers are strongly neurofilamentpositive, whereas satellite cells lack any detectable neurofilament. In B, in contrast, CD56 is expressed on the cell surface and on satellite cells of ganglion cells. In C,
CD57 is confined to myelin sheaths of thick nerve fibers present in an adjacent minor peripheral nene trunk (arroH's) and to scattered lymphoid cells, x 181.
Damaged Striated Muscle Cells. The great majority of striated
muscle fibers microtopographically associated with malignant
tumor cells were CD56+ in all seven cases studied (Fig. &4), as
cells in one of two cases. In one skeletal muscle which showed
extensive fibrosis due to previous operative intervention, most
of the muscle cells were CD56+ (Fig. 8Ä),as were muscle fibers
were most of the muscle fibers associated with benign tumor
of a skeletal muscle biopsy specimen which showed necrosis
caused by vascular embolization. Like in normal striated muscle
cells, CD57 was absent in all damaged striated muscle cells.
Tumors of Striated Muscle Origin. Included were six em
bryonal, one alveolar, and one pleomorphic RMS. In four
embryonal, in the alveolar, and in the pleomorphic RMS, all
tumor cells, i.e., small, round, undifferentiated tumor cells as
well as rounded or elongated rhabdomyoblasts present in var
ious amounts, were strongly and consistently CD56+ (Fig. 9A).
In one embryonal RMS a major tumor cell population ex
pressed CD56 molecules; in another case, which was exclusively
composed of rounded rhabdomyoblasts, a minor tumor cell
population also expressed CD56 molecules. All RMS studied
were CD57~.
DISCUSSION
There is increasing evidence for mAbs to detect identical
epitopes that might be part of different structures. This holds
true for some of the CD antigens which may be found in various
normal and/or neoplastic extrahematopoietic cells, either constitutively or resulting from malignant transformation.
CD57(HNK-1) is one of the best studied antigens. Outside the
hematopoietic system it has been found in a series of neural or
neuroectodermal tumors, including neurofibromas and neurofibrosarcomas (27, 28), malignant melanomas (29, 30), malig
nant peripheral neuroectodermal tumors (30, 31 ), Ewing's sar
Fig. 4. Traumatic neuroma (ischiadic nerve). In A. nearly all hapha/ardly
oriented nerve fibers express CD56 molecules. In B, in contrast to the normal
state, some of the CD56-positive nerve fibers also show CD57-positive myelin
sheaths. X 145.
comas (32), and even small cell lung carcinomas and other
neuroendocrine tumors (33). This seems to reflect the fact that
CD57 also recognizes myelin-associated glycoprotein and, ad
ditionally, a sulfated carbohydrate epitope expressed on a por
tion of N-CAM isoforms (5).
CD56(NKH-1), which belongs also to the NK-associated
antigens, has recently been demonstrated to be identical to the
M, 140,000 isoform of N-CAM (8). This prompted us to study
the expression of CD56 and CD57 in human neural and striated
muscle cells as well as in their tumors by application of mAbs
Leu-19(8)andLeu-7(l).
Our results on the expression of CD56 and CD57 antigens
in normal human nerve fibers are in parallel with data reported
by Nieke and Schachner (34) who, in adult rats, found N-CAM
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CD56 AND CD57 [N HUMAN NEURAL AND STRIATED MUSCLE TISSUES
Fig. 5. A, spindle cell malignant schwannoma of the buttock. CD56 is detectable in the entire tumor cell population. B, pleomorphic malignant schwannoma of
the retroperitoneum. About one-half of the tumor cell population lacks any detectable CD56 molecules. C, spindle cell malignant schwannoma of the stomach. The
tumor cells are CD57-negative throughout. Scattered CD57-positive lymphoid cells serve as an intrinsic control for the immune reaction, x 181.
Fig. 6. /i, ganglioneuroblastoma of the adrenal gland. Small, round, undifferentiated neuroblasts, scattered immature ganglion cells (arrows), and neuropillike processes show strong expression of CD56 antigens. Additionally, extracel
lular deposits of CDS6 are found around larger neuroblasts (stars), x 145. B,
neuroblastoma of the thoracic wall. The entire undifferentiated tumor cell popu
lation is strongly CDSo-positive, while endothelial cells are unstained, x 181.
immunoreactivity predominantly associated with unmyelinated
Schwann cells, whereas the CD57(HNK-1) epitope appeared to
be associated with the outer profile of thick myelin sheaths, a
finding which was also been described by Smolle et al. (27) who
investigated the pattern of CD57 expression in a variety of skin
tumors. In traumatic neuromas and in the Schwann cell com
partment of ganglioneuromas,
however, CD56 was more
broadly distributed and could also be found in some CD57positive Schwann cells. In the distal end of transected rat
ischiadic nerves, N-CAM also reappeared in many Schwann
cells (34-36); however, N-CAM was restricted to CD57-negative nerve fibers. Like in traumatic neuromas and in the
Schwann cell compartment of ganglioneuromas, CD56 was
consistently expressed in benign schwannomas whereas, in con
trast to data, CD57 and/or neurofilament were only rarely
detectable in these neoplasms and in their malignant counter
parts (27, 28). Malignant schwannomas, however, exhibited a
heterogenous pattern of CD56 expression, i.e., some cases were
partially or even completely CD56-negative, reflecting the ab
sence of at least the M, 140,000 isoform of N-CAM in these
tissues. Since N-CAM is required for normal adhesive interac
tions between cells, low amounts or even absence of N-CAM
might lead to a loss of contact inhibition allowing cell detach
ment and migration in an uncontrolled manner. In line with
this hypothesis, Linnemann et al. (37) recently demonstrated
that the metastatic melanoma cell line K1735-M1 expressed
less N-CAM than did the nonmetastasizing K1735-C116 line.
Furthermore, mAb MUC18, a marker for tumor progression
in human melanoma, has been found to show sequence similar
ity to neural cell adhesion molecules including N-CAM (38).
Additionally, N-CAM expression is markedly reduced after
transformation of retinal and neural cells with Rous sarcoma
virus (39, 40).
N-CAM expression in neuroblastomas was initially described
in mice showing a shift to the predominant expression of the
A/r 180,000 isoform in more differentiated neuroblasts (41).
Since the M, 180,000 isoform of N-CAM occurs at later stages
during development than do the M, 120,000 and 140,000
isoforms, it is suggested that A/r 180,000 N-CAM plays a
differential role in the stabilization of cell contacts. N-CAM
expression in human neuroblastoma cell lines was first de-
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CD56 AND CD57 IN HUMAN NEURAL AND STRIATED MUSCLE TISSUES
:7C •¿
v
.-
Fig. 7. In A, a fetal skeletal muscle of 18 weeks of gestation exhibits strong surface expression of CD56 in all muscle fibers. B and C, serial section preparation of
an adult skeletal muscle. In B, the muscle fibers lack any detectable CD56. Intrafusal muscle fibers of a muscle spindle are at least partially CDSo-positive, as are
some small nerve fibers. In C, CD57 is confined to myelin sheaths of CDSo-negative nerve fibers, x 181.
scribed by Lipinski et al. (42) using a rabbit anti-human
N-CAM antiserum. We found expression of CD56 in all (ganglio)neuroblastomas studied. Feickert et al. (43), using the same
antibody we applied, found in situ expression of M, 140,000 N-
CAM in only 6 of 11 neuroblastomas, whereas CD56 mAb T199 yielded positive results in all cases studied. Since CD56
mAb only detects the M, 140,000 isoform of N-CAM, the
CD56 negativity of some neuroblastomas might reflect differ
ences in the maturation grade of the various neoplasms studied.
However, our cases, which included different grades of matu
ration, were all CD56 positive. In contrast, CD56 was only
rarely detectable in peripheral neuroepitheliomas. Since NCAM has been found in a series of Ewing's sarcomas both in
situ and in vitro (42, 44, 45) and since Ewing's sarcoma and
8B
Fig. 8. A, peripheral neuroepithelioma of the thoracic wall. The undifferentiated tumor cells are consistently CD56-negative, whereas infiltrated skeletal
muscle fibers are strongly CD56-positive. x 181. A, skeletal muscle of the lower
extremity showing extensive fibrosis due to previous operative intervention. The
great majority of muscle fibers are strongly CD56-positive. x 145.
peripheral neuroepithelioma are considered to be related neo
plasms based on the shared reciprocal t(ll;22) translocation
(46) and the shared protooncogene expression (47), the rare
CD56 expression in peripheral neuroepitheliomas is notewor
thy. Another remarkable feature was the consistent absence of
CD57 in all (ganglio-)neuroblastomas studied. This contrasts
with data by Caillaud et al. (30) who found CD57 expression
in 11 of 13 neuroblastomas studied. However, since in our
hands some CD57-positive lymphoid cells and/or thick nerve
fibers showed reliable immune reaction and since mAb Leu-7
is of IgM isotype, the considerable number of CD57-positive
neuroblastomas reported in the literature might at least partially
be the result of unspecific isotype effects which, e.g., can be
found in several epithelial cell types (data not shown).4
The knowledge of the distribution pattern of N-CAM in
human tissue is further enriched by results obtained with some
mAbs of the neuroblastoma and the small cell lung cancer
workshops, which have been found to also share an epitope in
common with N-CAM. Thus, mAbs UJ13A and 3F8 have been
demonstrated in situ in 4 of 4 neuroblastomas and in vitro in 6
of 8 neuroblastoma cell lines, respectively (45, 48). In addition
to sharing an epitope in common with N-CAM, mAb 3F8 also
recognizes the ganglioside GD2 (49, 50) which appears to be a
particularly good marker for neuroectodermal tumors such as
neuroblastoma and malignant melanoma. Both UJ13A and 3F8
have gained diagnostic and/or therapeutic application (51-53).
Given that N-CAM is present in normal peripheral nerves and
autonomie ganglia, the diagnostic and therapeutic value of
mAbs recognizing an epitope in common with N-CAM should
be reassessed. In addition to their expression in neuroblastomas
and malignant melanomas, some of the mAbs sharing an epi
tope in common with N-CAM including CD56 mAb have also
been demonstrated in medulloblastomas, Ewing's sarcomas,
4 P. Möllerand K. Koretz, unpublished observations.
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CD56 AND CD57 IN HUMAN NEURAL AND STRIATED MUSCLE TISSUES
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. .
associated with tumor cells. All in all, the distribution pattern
of CD56 in neural and striated muscle cells and, according to
previous and recent literature, in their tumors corresponds to
that of N-CAM. CD57, in contrast, shows a pattern of expres
sion which in most instances is complementary to that of CD56
and thus far from covering that of N-CAM. The absence of
CD56 in some malignant schwannomas and peripheral neuroepitheliomas might reflect alterations in cell-cell interactions.
CD56 expression in normal fetal and regenerative striated
muscles, as well as in rhabdomyosarcomas, suggests a role of
this molecule as an oncodevelopmental antigen, since it is
absent in normal adult striated muscles. These results obtained
in striated muscle cells implicate parallels to those found in
renal tissue. Since N-CAM is expressed in normal peripheral
nerves and autonomie ganglia, the diagnostic and therapeutic
value of mAbs sharing an epitope in common with N-CAM has
to be reassessed.
. ««••.1*;.'
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ACKNOWLEDGMENTS
The skillful technical assistance of Inge Brandt and Sybille Menges
and the excellent photographic assistance of John Moyers are gratefully
acknowledged. We are indebted to Christine Raulfs for editing the
manuscript.
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Fig. 9. A, embryonal rhabdomyosarcoma of the retroperitoneum. Small, round,
undifferentiated tumor cells as well as rounded or elongated rhabdomyoblasts
display strong staining for CD56. Myxoid matrix is unstained, x 145. B, malig
nant schwannoma with rhabdomyoblastic differentiation (Triton tumor) of the
lower extremity. The majority of tumor cells, including some with rhabdomyo
blastic differentiation (arrows), are CD56 positive. xl81.
Wilms' tumors, and even in rhabdomyosarcomas
(42-45, 48).
In line with these results, Roth et al. (54) found evidence of the
long chain form of polysialic acid of N-CAM in Wilms' tumors.
Since N-CAM is present in fetal but absent in adult kidneys,
the authors suggested that this molecule might represent an
oncodevelopmental antigen in renal tissue. Like mAbs of the
neuroblastoma and small cell lung cancer workshops, CD56
was expressed in rhabdomyosarcomas, whereas CD57 was com
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to the reports of Feickert et al. (43), who showed in situ
expression of CD56 in 2 of 6 rhabdomyosarcomas only, we
found CD56 expression in variable amounts in 8 of 8 rhabdo
myosarcomas studied. Because N-CAM is present in fetal but
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reexpressed in rhabdomyosarcomas, we suggest that, in striated
muscle tissue, this antigen might also be considered an oncod
evelopmental antigen. Reexpression of N-CAM in regenerative
muscle fibers in various myopathies (55, 56) is in line with this
conclusion because regenerative muscle fibers also follow the
developmental pathway. Additionally, we found reexpression
of CD56 in regenerative muscle fibers in damaged skeletal
muscles and in skeletal muscle fibers microtopographically
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1307
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Expression of the Natural Killer Cell-associated Antigens CD56
and CD57 in Human Neural and Striated Muscle Cells and in
Their Tumors
Gunhild Mechtersheimer, Martina Staudter and Peter Möller
Cancer Res 1991;51:1300-1307.
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