B .
J.
Cance
(1987),
55,
421-426
©
The
Macmillan
P ess
L d.,
1987~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~-
Cha ac e iza ion
o
Fac o
XIII
con aining-mac ophages
in
lymph
nodes
wi h
Hodgkin's
disease
R.
Adany1,
Z.
Nemes2
&
L.
Muszbek'
'Depa men
o
Clinical
Chemis y
and
2Depa men
o
Pa hology,
Uni e si y
School
o
Medicine,
Deb ecen,
H-4012,
Hunga y.
Summa y
A
la ge
numbe
o
cells
con aing
subuni
a
o
blood
coagula ion
Fac o
XIII
(FXIII)
was
de ec ed
by
immunope oxidase
s aining
in
lymph
nodes
wi h
Hodgkin's
disease.
These
ela i ely
la ge,
mul ipola ,
mononuclea
cells
we e
o en
ound
in
he
immedia e
icini y
o
malignan
Hodgkin's
cells.
In ensi e
cha ac e iza ion
o
hese
cells
ca ied
ou
by
immuno luo escen
and
enzymecy ochemical
echniques
in
double-
and
iple-labelling
sys ems
on
he
same
sec ions
clea ly
demons a ed
ha
hey
ep esen
umou -
associa ed
mac ophages
(TAMs).
FXIII
con aining-cells
showed
a-naph yl
ace a e
es e ase
(ANAE)
posi i i y,
and
we e
labelled
by
monoclonal
an i-Leu
M3
an ibody,
a
monocy e/mac ophage
ma ke ,
bu
no
a
all
o
only
e y
weakly
by
an i-HLA-DR.
Nei he
alkaline
phospha ase
(ALP)
no
adenosine
iphospha ase
(ATPase)
ac i i y
could be
de ec ed
in
hese
cells
and
su p isingly,
hey
we e
consis en ly
nega i e
o
acid
phospha ase
(AcP)
as
well.
The
p esence
o
FXIII
subuni
a
in
umou -associa ed
mac ophages
sugges s
ha
his
cell
ype
migh
ha e
an
impo an
ole
in
he
s abiliza ion
o
ib in
deposi s
a ound
umou
cells.
Fib in
deposi ion
is
a
a he
equen
inding
in
spon aneously-a ising
as
well
as
ansplan able
human
and
animal
umou s
and
has
been
implica ed
in
a ious
aspec s
o
umou
g ow h
and
me as asis
(Rickles
&
Edwa ds,
1983;
D o ak
e
al.,
1983).
Though
mos
o
pe haps
all
malignan
cells
possess
p ocoagulan
ac i i ies
(O'Mea a,
1958;
Go don
e
al.,
1975;
1979;
Go don
&
C oss,
1981;
Seme a o
&
Dona i,
1981)
ha
can
ac i a e
he
coagula ion
cascade,
i
is
becoming
inc easingly
e iden
ha
umou -associa ed
mac o-
phages
(TAMs)
a e
also
in ol ed
in
in a- umo al
ib in
o ma ion
(Edwa ds
e
al.,
1981;
E ans,
1982;
Lo enze
e
al.,
1983;
Key,
1983).
Mac ophages
o ,
in
iew o
hei
he e ogenei y
(Hoppe
e
al.,
1979;
Poul e
e
al.,
1983),
ce ain
o
hei
subse s
could
con ibu e
o
ex a ascula
clo ing
by
wo
mechanisms.
(i)
By
exp essing
issue
ac o
ac i i y
hey
can
ini ia e
he
ex insic
coagula ion
pa hway
(Rickles
&
Edwa ds,
1983;
D o ak
e
al.,
1983;
Lo enze
e
al.,
1983).
(ii)
They
con ain
a
numbe
o
clo ing
ac o s
which,
i
sec e ed
o
eleased
om
damaged
cells
in o
he
in e s i ial
space,
p o ide
all
he
componen s
necessa y
o
ex insic
h ombin
o ma ion.
The
p oduc ion o
i amin
K-dependen
clo ing
ac o s
(Fac o
II,
VII,
IX,
X)
and
Fac o
V
by
mac ophages
has
been
well
es ablished
(Os e ud
e
al.,
1980;
Lindahl
e
al.,
1982;
an
Dam-Mie as
e
al.,
1985;
Chapman
e
al.,
1985).
Mos
ecen ly
he
p esence
o
subuni
a
o
Fac o
XIII
(FXIII),
he
enzyma ically-ac i e
cons i uen
o
ib in
s abilizing
ac-
o
has
also
been
demons a ed
in
human
pe iphe al
blood
monocy es
(Muszbek
e
al.,
1985)
and
pe i oneal
mac o-
phages
(Adany
e
al.,
1985)
in
ou
labo a o y.
Independ-
en ly,
hese
esul s
ha e
been
con i med
by
Hen iksson
e
al.
(1985).
FXIII
deli e ed
by
mac ophages
in o
he
umou
s oma
migh
ha e
an
impo an
ole
in
he
s abiliza ion
o
ib in
o med
ex a ascula ly
and
in
i s
ac i a ed
o m
as
a
ansglu aminase
i
migh
exe
o he
biological
unc ions
as
well.
Thus,
i
was
in e es ing
o
see
i
TAMs
o
ce ain
subse s
o
hem
con ain
FXIII.
The
p esence
o
ib in
deposi s
in
lymph
nodes
wi h
Hodgkin's
disease
is
a
gene al
obse a ion
(Ha is
e
al.,
(1982;
D o ak
e
al.,
1983)
and
in
hese
lymph
nodes
mac ophage-like
cells
p e ail
o e
all
o he
cell
ypes
in
a eas
ich
in
ib illa
in e cellula
subs ance
(S ille
&
Ka enkamp,
1978;
Hansmann
&
Kaise ling,
1981).
He e
we
show
ha
a
dis inc
cell
popula ion
in
lymph
nodes
wi h
Hodgkin's
disease
con ains
subuni
a
o
FXIII.
The
mac ophage
na u e
o
hese
cells
is
clea ly
demons a ed
and
by
using
double-
Co espondence:
R.
Adany.
Recei ed
21
July
1986;
and
in
e ised
o m,
10
No embe
1986.
and
iple-labelling
echniques
he
FXIII
con aining
subse
o
TAMs
is
ex ensi ely
cha ac e ized.
T iple
labelling
sys ems
p o ide
an
excellen
oppo uni y
o
ca y
ou
an
exac
cha ac e iza ion
o
a
cell
popula ion,
because
he
di ec
demons a ion
o
wo
di e en
an igens
in
combina ion
wi h
enzyme-cy ochemical
eac ions
in
he
same
sec ion
yields
a
p ecise
ep esen a ion
o
he
coincidence
o
h ee
di e en
cha ac e is ics.
Ma e ials
and
me hods
Lymph
node
biopsies
we e
ob ained
om
12
pa ien s
wi h
Hodgkin's
disease
o
nodula
scle osing
ype.
Sec ions
om
non-neoplas ic,
eac i e
lymph
nodes
se ed
as
con ols.
All
specimens
we e
di ided
in o
wo
pa s
a
he
ime
o
su gical
biopsy.
One
pa
was
ixed
in
3.5%
pa a o maldehyde
ixa i e
(4h,
oom
empe a u e)
hen
acuum
embedded
in
pa a in
and
sec ioned
in o
6pim
slides,
while
he
o he
pa
was
snap- ozen
and
cu
in
a
c yos a .
Six
m
ozen
sec ions
we e
ai -d ied,
w apped
in
oil
and
s o ed
a
-20°C.
Immunope oxidase
s aining
Fo maldehyde- ixed
pa a in
embedded
sec ions
we e
de-
waxed
and
ehyd a ed.
Endogenous
pe oxidase
ac i i y
was
blocked
by
I
%
H202
in
absolu e
me hanol
o
30
min
a
oom
empe a u e.
Sec ions
we e
diges ed
wi h
0.1
%
ypsin,
in
TRIS-bu e ed
saline
(pH
7.6)
con aining
0.1
%
calcium
chlo ide
a
37DC
o
20min.
Non-speci ic
IgG
binding
was
p e en ed
by
p eincuba ion
wi h
20%
no mal
goa
se um
o
15
min.
Sec ions
we e
co e ed
o
2
h,
a
oom
empe a u e
wi h
abbi
an ise um
agains
FXIII
subuni
a
(Beh ingwe ke
AG,
Ma bu g,
Wes
Ge many)
dilu ed
1:25
wi h
20%
no mal
goa
se um.
The
monospeci i y
o
his
an ise um
was
e i ied
by
immunoblo ing
on
whole
human
plasma
as
well
as
on
human
pla ele
and
monocy e
homogena e
(Muszbek
e
al.,
1985).
An igen-an ibody
eac ion
was
de ec ed
by
bio inyla ed
an i- abbi
IgG
and
a idin-bio inyla ed
pe -
oxidase
complex
(Vec as ain
ABC
ki )
(Vec o
Labo a o ies,
Bu lingame,
CA).
The
speci ic
pe oxidase
ac i i y
was
isualized
by
0.05%
3,3'-diaminobenzidine
e ahyd o-
chlo ide
(DAB)
(Sigma
Co.,
S .
Louis,
MO),
0.01%
H202
in
0.1
mol
1-
1
TRIS
HCI
bu e ,
pH
7.2.
Coun e s aining
was
wi h
Maye 's
haema oxylin
be o e
dehyd a ion
in
g aded
alcohol
and
moun ing
wi h
Canada
balsam.
On
con ol
slides
no mal
abbi
se um
a
he
same
dilu ion
was
used
ins ead
o
an i
FXIII
subuni
a
an ise um.
PBS,
pH
7.3,
was
used
o
an ibody
dilu ion
and
in
washing
p ocedu es.
B .
J.
Cance
(1987),
55,
421-426
,'-.
The
Macmillan
P ess
L d.,
1987
422
R.
ADANY
e
al.
Immuno luo escen
and
enzyme-his ochemical
echniques
F ozen
sec ions
we e
used
o
he
double
immunolabelling
echniques
combined
wi h
he
cy ochemical
localiza ion
o
se e al
enzymes
on
he
same
slide
(combined
iple
labelling
sys ems).
Immedia ely
be o e
immuno eac ions
c yos a
sec-
ions
we e
unw apped
and
ixed
in
ace one
a
+4°C,
o
10
min.
A e
washing
in
PBS,
slides
we e
incuba ed
wi h
1:200
dilu ion
o
an ise um
agains
FXIII
subuni
a
o
2
h
a
oom
empe a u e.
As
he
seconda y
an ise um,
a
1:40
dilu ion
o
swine
an i- abbi
IgG
luo esceina ed
(Dakopa s
a/s,
Glos up,
Denma k),
was
used
(30
min
incuba ion).
In
he
nex
s age
his
eac ion
was
combined
ei he
wi h
he
de ec ion
o
HLA-DR
an igen
o
wi h
he
isualiza ion
o
Leu
M3,
a
monocy e/mac ophage
su ace
ma ke .
Sec ions
we e
incuba ed
wi h
1:5
dilu ion
o
bio inyla ed
mouse
an i-
human
HLA-DR
monoclonal
an ibody
(Bec on-Dickinson,
Sunny ale,
CA)
o
wi h
he
same
dilu ion
o
mouse
an i-
human
Leu
M3
monoclonal
an ibody
conjuga ed
wi h
phyco-
e y h in
(Bec on-Dickinson,
Sunny ale,
CA)
o
30
min,
a
oom
empe a u e.
The
speci ic
binding
o
bio inyla ed
an i-
HLA-DR
an ibody
was
de ec ed
by
1:40
dilu ion
o
s ep a idin-Texas
ed
(Ame sham,
UK).
In
he
case
o
nega i e
con ols
non-immune
abbi
se um
and
con ol
mouse
IgG
om
umou -bea ing
BALB/c
mice
conjuga ed
wi h
phycoe y h in
o
FITC
(Bec on
Dickinson,
Sunny ale,
CA)
we e
subs i u ed
o
he
i s
an ibodies.
Following
double
immuno luo escen
s aining,
sec ions
we e
moun ed
in
50%
glyce ol
in
PBS
and
examined
unde
an
Op on
ul a iole
mic oscope
equipped
wi h
an
epi luo escence
condenso
con aining
selec i e
il e s
o
FITC
and
Texas
ed/phycoe y h in.
A e
pho og aphs
had
been
aken,
co e -slips
we e
emo ed,
sec ions
we e
washed
ho oughly
in
dis illed
wa e .
As
a
hi d
s ep,
one
o
he
ollowing
enzymes
was
de ec ed:
oc-nap hyl
ace a e
es e ase
(ANAE)
(Muelle
e
al.,
1975),
acid
phospha ase
(AcP)
(Poul e
e
al.,
1983),
aden-
osine
iphospha ase
(ATPase)
(Poul e
e
al.,
1983)
and
alkaline
phospha ase
(ALP)
(Mason
&
Wools on,
1982).
In
he
case
o
nega i e
con ol
slides
in
he
enzymecy o-
chemical
eac ions
ei he
he
subs a e
was
omi ed
om
he
incuba ion
medium,
o
be o e
de elopmen
o
an
enzyme
eac ion
he
app op ia e
enzyme
ac i i y
was
blocked
acco d-
ing
o
he
ecommenda ion
o
Poul e
e
al.
(1983).
Finally,
sec ions
we e
emoun ed
in
50%
glyce ol
in
PBS
and
he
ields
o
which
pho og aphs
had
been
p e iously
aken
we e
iden i ied
in
no mal
ligh
mic oscope
and
epho og aphed.
Using
he
abo e-men ioned
immuno-
and
enzyme-his o-
chemical
eac ions
all
possible
combina ions
o
iple
label-
ling
we e
pe o med
on
e e y
biopsy
specimen.
Resul s
Immunope oxidase
s aining
on
pa a o maldehyde- ixed,
pa a in
embedded
sec ions
was
used
o
e i y
he
p esence
and
de e mine
he
dis ibu ion
o
FXIII
subuni
a
con aining
cells
in
lymph
nodes
wi h
Hodgkin's
disease
o
nodula
scle osing
ype.
As
demons a ed
in
Figu es
la,
4b,
5a
and
6a
p ac ically
he
whole
a ea
o
lymph
node
was
in il a ed
by
cells
showing
in ensi e
s aining
o
FXIII.
These
cells
we e
o en
ound
in
he
immedia e
icini y
o
malignan
Hodgkin's
cells
sugges ing
an
in ima e
ela ionship
be ween
he
wo
cell
ypes
(Figu e
lb).
FXIII
con aining-cells
in
lymph
nodes
wi h
Hodgkin's
disease
possess
a
mac ophage-like
appea ance,
hey
a e
ela -
i ely
la ge,
mul ipola ,
mononuclea
cells
wi h
nume ous
acuoles
in
he
cy oplasm.
Thei
de ailed
cha ac e iza ion
was
pe o med
by
iple
labelling
(double
immuno-
luo escen
+
enzyme-cy ochemical)
echniques.
S aining
o
FXIII
and
he
monocy e/mac ophage
ma ke
Leu
M3
(Dimi iu-Bona
e
al.,
1983)
showed
an
iden ical
dis ibu ion
pa e n
in
each
case
(Figu e
3a,
b).
This
ac
oge he
wi h
hei
de ini e
ANAE
posi i i y
(Figu e
4b,
c)
clea ly
iden-
i ies
FXIII
con aining-cells
as
membe s
o
he
mac ophage
amily.
These
cells
we e
p edominan ly
nega i e
o
HLA-
DR
hough
occasionally
e y
weak
posi i e
s aining
could
also
be
de ec ed
(Figu es
2,
4a,
b).
ALP
(Figu e
5a, b)
and
ATPase
ac i i ies
(no
shown)
could
no
be
de ec ed
in
FXIII
con aining
cells
and
su p isingly,
hey
we e
consis -
en ly
nega i e
o
AcP
(Figu e
6a,
b),
as
well.
The
p esence
o
HLA-DR
o
AcP
posi i e
bu
FXIII
nega i e
cells
wi h
mac ophage-like
mo phological
ea u es
clea ly
indica es
ha
FXIII
is
exp essed
only
in
a
ce ain
subse
o
TAMs.
In
con as
o
lymph
nodes
wi h
Hodgkin's
disease,
in
eac i e
lymph
nodes,
FXIII
con aining
cells
we e
localized
almos
exclusi ely
in
pe i ascula
connec i e
issue
and
in
subcapsula
o
medulla y
sinuses.
Mac ophages
in
ollicles
o
eac i e
lymph
nodes
iden i ied
by
s ong
ANAE
eac ion
we e
consis en ly
nega i e
o
FXIII
subuni
a
and
only
a
ew
posi i e
cells
could
be
de ec ed
in
he
in e ollicula
a eas
(no
shown).
A
de ailed
cha ac e iza ion
o
FXIII
con aining-cells
in
eac i e
lymph
nodes
is
published
elsewhe e
(Nemes
e
al.,
1986).
Discussion
Plasma
FXIII,
like
mos
o he
clo ing
ac o s
ci cula es
as
a
zymogen.
I is
a
e ame ic
p o ein
consis ing
o
wo
ypes
o
subuni s
(a2b2).
The
po en ial
enzyma ic
si e
is
loca ed
on
subuni
a
which
can
assume
an
ac i e
con igu a ion
only
ollowing
p o eoly ic
clea age
by
h ombin
and
Ca2
+
in-
duced
dissocia ion
om
he
inhibi o y
b
subuni
(see
Muszbek
&
Laki,
1984
o
e iew).
I
has
been
known
o
a
long
ime
ha
subuni
a
bu
no
b
also
exis s
as
an
in acellula
p o ein
in
pla ele s
(Buluk,
1955),
mega-
ka yocy es
(Kiesselbach
&
Wagne ,
1972)
and
placen a
(Bohn
&
Schwick,
1971).
As
men ioned
ea lie
he
exis ence
o
subuni
a
in
monocy es
has
also
been
e i ied
(Muszbek
e
al.,
1985)
and
hese
cells
e ain
hei
FXIII
con en
ollowing
di e en ia ion
in o
pe i oneal
mac ophages
(Ad'any
e
al.,
1985).
Fu he mo e,
i
was
shown
ha
FXIII
con aining
cells
in
he
placen a
a e
also
o
mac ophage
o igin
(Adany
&
Muszbek,
submi ed
o
publica ion).
In
his
s udy
a
cell
ype
exp essing
FXIII
subuni
a
was
de ec ed
in
lymph
nodes
wi h
Hodgkin's
disease.
The
mo pho-
logical
appea ance
o
FXIII
con aining
cells
indica ed
ha
hey
belong
o
TAMs,
hus
combina ions
o
immuno-
and
enzyme-his ochemical
eac ions
-
gene ally
used
o
iden i i-
ca ion
and/o
pheno yping
o
mac ophages
-
we e
applied
o
cha ac e ize
hem.
Thei
mac ophage
na u e
was
clea ly
demons a ed
by
Leu
M3
and
ANAE
posi i i y.
A
he
same
ime
nega i e
eac ions
we e
ound
o
HLA-DR,
AcP,
ALP
and
ATPase.
The
abo e
esul s
s ongly
sugges
ha
his
FXIII
con aining-TAM
cell
ype
is
no
iden ical
wi h
any
o
he
mac ophage
cell
ypes
cha ac e ized
ea lie
in
no mal
human
lymph
node.
This
conclusion
is
clea ly
suppo ed
by
he
ollowing
da a:
(i)
FXIII
con aining-mac ophages
a e
no
iden ical
wi h
in e digi a ing
e icula
cells
and
den i ic
cells
o
lymph
node
because
he
la e s
a e
s ongly
posi i e
o
HLA-DR
and
ATPase
(Janossy
e
al.,
1980;
S ein
e
al.,
1980;
Poul e
e
al.,
1983).
(ii)
They
di e
om
sinus
his iocy es
o
no mal
lymph
node
and
om
in lamma o y
mac ophages,
as
well
as
by
he
absence
o
AcP
ac i i y
(Poul e
e
al.,
1983).
(iii)
Being
ALP
nega i e
hey
may
no
be
in e p e ed
as
ib oblas ic
e iculum
cells
(Chilosi
e
al.,
1981).
On
he
o he
hand,
FXIII
con aining-cells
in
lymph
nodes
wi h
Hodgkin's
disease
a e
no
e en
iden ical
wi h
o he
FXIII
con aining-cells
o
no mal
lymph
nodes
which
occu
in
he
connec i e
issue
o
capsule
and
in
he
sinus
(Nemes
e
al.,
1986).
HLA-DR
posi i i y
o
connec i e
issue
his iocy es
as
well
as
he
in ensi e
AcP
eac ion
o
sinus
mac ophages
clea ly
dis inguish
he
la e
cell
ypes
om
FXIII
con aining-TAMs.
FXIII
IN
TUMOUR
ASSOCIATED
MACROPHAGES
423
Figu e
1
Immunope oxidase
s aining
o
FXIII
subuni
a
combined
wi h
haema oxylin-ch omo ope
coun e s aining
on
a
sec ion
o
pa a o maldehyde- ixed,
pa a in
embedded
lymph
node
wi h
Hodgkin's
disease,
nodula
scle osis
ype.
FXIII
subuni
a-
con aining
cells
(in
b own
colou )
in aded
p ac ically
he
whole
a ea
o
lymph
node
(a)
and
we e
equen ly
ound
in
he
immedia e
icini y
o
malignan
Hodgkin's
cells
(b).
Ba =
25
gm
(a)
and
10pm
(b).
Figu e
2
Double
immuno luo escen
labelling
o
FXIII
subuni
a
and
HLA-DR
on
a
c yos a
sec ion
o
lymph
node
wi h
Hodgkin's
disease.
In
a
double
exposu e
pho og aph
g een
FXIII
con aining
cells
a e
easily
dis inguishable
om
Texas
ed
labelled
HLA-DR
posi i e
ones.
Ba
=10
lOm.
Figu e
3
In
lymph
node
wi h
Hodgkin's
disease
iden ical
cells
we e
labelled
wi h
immuno luo escen
s aining
o
FXIII
subuni
a
(a)
and
o
he
monocy e/mac ophage
su ace
an igen
Leu
M3
(b).
Ba =
10
pm.
D
424
R.
ADANY
e
al.
Al
C*
.
Figu e
4
T iple
labelling
o
HLA-DR
(a),
FXIII
subuni
a
(b)
and
ANAE
(c)
on
he
same
a ea
o
lymph
node
wi h
Hodgkin's
disease.
FXIII
con aining
cells
a e
ANAE
posi i e
(see
a ows),
bu
HLA-DR
nega i e.
Ba =
25
im.
Di e en
ypes
o
mac ophages
o igina e
om
a
common
bone
ma ow
p ecu so
( an
Fu h,
1981)
bu
owing
o
an
in ica e
di e en ia ion
p og am
associa ed
wi h
he
ex-
p ession
and
disappea ance
o
a ious
gene
p oduc s
(Dimi iu-Bona
e
al.,
1983)
hey
show
an
ex eme
pheno-
ypic
and
unc ional
di e si y.
In
conside a ion
o
his
ac
we
belie e
ha
FXIII
con aining-TAMs
a e
a
subse
o
mac ophages
di e en ia ed
om
blood
monocy es
o
special
unc ion(s)
ela ed
o
malignan
cell
p oli e a ion.
Indepen-
den ly
om
FXIII
con aining-mac ophages
some
AcP
posi i e
mac ophage-like
cells
we e
also
de ec ed.
This
ind-
ing
oge he
wi h
he
p esence
o
HLA-DR
posi i e,
bu
FXIII
negn-i i e
cclls
in
lymph
nodes
wi h
Hodgkin's
diseaise
suppo
he
possibili y
ha
TAMs
a e
o
be
conside ed
as
an
inhomogeneous
cell
popula ion.
Ex a ascula
ib in
deposi ion
occu s
in
he
s oma
and
a
hos - umou
in e ace
in
mos
o
pe haps
all
malignan
neoplasms
(D o ak
e
al.,
1983).
The
pa hological
signi-
icance
and
he
o igin
o
ib in
deposi s
has
no
been
su icien ly
explo ed.
Mos
o
he
a ailable
clinical
and
expe imen al
da a,
howe e ,
end
o
suppo *
he
iew
ha
he
ac i a ion
o
clo ing
sys em
is
bene icial
o
bo h
umou
p og ession
and
me as asis
o ma ion.
Mos ly
on
he
basis
o
heo e ical
conside a ions
he
ollowing
hypo heses
ha e
been
p oposed
o
he
pa hogenic
ole
o
ib in
o med
be ween
and
a ound
umou
cells:
(1)
i
migh
ha e
a
ba ie
unc ion
and
in e e es
wi h
he
hos 's
immune
esponse,
(2)
i
could
s imula e
umou
angiogenesis,
(3)
i
may
ha e
a
ole
in
he
implan a ion
o
ci cula ing
umou
cells
a
me as a ic
si es
(D o ak
e
al.,
1983).
An
impo an
ole
o
FXIII
-
ib in
s abilizing
ac o
-
in
any
o
he
abo e
mechanisms
seems
a he
ob ious.
By
o ming
ib inolysis-
esis an
ib in
meshwo k
and
c osslinking
ibn lla
in e -
cellula
ma ix
componen s,
FXIII
migh
suppo
he
ba ie
unc ion
o
migh
e en
be
essen ial
o
i .
The
p ocess
o
angiogenesis
and
ib oplasia
du ing
umou
g ow h
and
wound
healing
has
been
compa ed
(D o ak
e
al.,
1983)
and
i
is
well
known
ha
wound
healing
is
highly
impai ed
in
FXIII
de icien
pa ien s
(Ducke ,
1972).
A
possible
di ec
e ec
o
FXIII
on
cell
p oli e a ion,
as
obse ed
in
he
case
o
ib oblas s
(Beck
e
al.,
1961),
migh
also
ha e
some
implica ions
o
umou
g ow h.
A
u he
possibili y
con-
ce ns
he
ansglu aminase
na u e
o
ac i a ed
FXIII.
As
a
ansglu aminase
i
can
a ach
hos
p o eins
co alen ly
o
he
memb ane
o
umou
cells
and
mask
hei
pu a i e
'non-sel '
cha ac e
esul ing
in
inc eased
immune
esis ance.
The
la e
idea
is
suppo ed
by
da a
showing
ha
a achmen
o
ib inogen
o
he
memb ane
o
YPC-1
plasmocy oma
cells
by
issue
ansglu aminase
esul s
in
an
inhibi ion
o
cell
media ed
cy o oxic
esponse
(Hunyadi
e
al.,
1981).
In
umou s
FXIII
could
ge
in o
he
in e s i ial
space
om
wo
possible
sou ces.
I
may
leak
om
blood
essels
oge he
wi h
o he
plasma
p o eins
due
o
enhanced
mic o ascula
pe meabili y
o
i
may
o igina e
om
he
TAM
subg oup
desc ibed
he e
by
ac i e
sec e ion
and/o
elease
ollowing
cell
des uc ion.
Howe e ,
his
ques ion
has
no
been
ad-
d essed
expe imen ally.
Clea ly,
u he
in es iga ions
a e
o
be
ca ied
ou
o
es
he
abo e
hypo heses
and
es ablish
he
ole
o
FXIII
as
well
as
FXIII
con aining-TAMs
in
he
p og ession
o
malignan
umou s.
These
s udies
we e
conduc ed
in
pa
pu suan
o
a
con ac
wi h
he
Na ional
Founda ion
o
Cance
Resea ch,
Be hesda,
Ma yland,
USA.
We
hank
M
R6be
He endi
o
he
expe
echnical
assis ance
and
M s
Mi ia
Kozma
o
he
excellen
sec e a ial
wo k.
FXIII
IN
TUMOUR
ASSOCIATED
MACROPHAGES
425
Figu e
5
Immuno luo escen
s aining
o
FXIII
subuni
a
(a)
combined
wi h
he
de ec ion
o
alkaline
phospha ase
(b)
on
he
same
sec ion
o
lymph
node
wi h
Hodgkin's
disease.
As
alkaline
phospha ase
ac i i y
was
e ealed
wi h
naph ol
AS-MX
plus
Fas
Red
TR
he
end
p oduc
o
he
eac ion
appea ed
in
ed
colou
no
only
in
no mal
ligh
bu
on
he
FITC
channel
o
he
luo escence
mic oscope,
as
well.
On
he
pho og aph
i
is
supe imposed
on
FITC
labelled
immun eac ion
o
FXIII
subuni
a.
FXIII
con aining
mac ophages
a e
nega i e
o
alkaline
phospha ase.
Ba =
25
pm.
Figu e
6
In
lymph
node
wi h
Hodgkin's
disease
immuno luo escen
s aining
o
FXIII
subuni
a
(a)
and
acid
phospha ase
ac i i y
de ec ion
(b)
isualize
wo
dis inc
cell
popula ions.
Ba =25pm.

426
R.
ADANY
e
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