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Prothrombin
Complex
Proteins
Platelet
Aggregation.
I. Inhibition
by Antiserum
By John
B. Miale
and
Data are presented
to support
the concept
that the vitamin
K-dependent
coagulation
factors
are
adsorbed
onto
the platelet
membrane
and
that
these make
up part
of the “plasma
atmosphere”
necessary
for
the aggregation
of platelets
by various
agents.
Together
with evidence
that other
coagulation
factors
also are part
of the
plasma
atmosphere,
it is suggested
that
T
HE ROLE
investigated
OF
in platelet-rich
vitro studies58
cofactor.
special
for platelet
aggregation
has been
platelet
aggregation
is diminished
platelet
aggregation
in
to aggregate
which
then
may
the
vitamin
these
be the
von
plus
in
as a
Willebrand’s
platelets’#{176}-” can
plasmatic
be
cofactor
in
circumstance.
The
data
factors
II,
sorbed
normal
onto
the
platelets
thrombin
VIII,’2
Kent
the aggregation
reaction
is part of the
coagulation
sequence.
The immunologic
approach
to demonstrating
constituents
of
the platelet
membrane
promises
to be a
highly
specific
technique
for
studying
further
the constituents
of the platelet
membrane
and their reactions
in hemostasis.
of ristocetin
factor
W.
in
with congenital
afibrinogenemia’4
fibrinogen,
plasmatic
or intracellular,9
of
failure
by adding
very
from
subjects
attention
on
that
Jessie
cofactors
reports
that
investigations
indicate
corrected
this
plasma
focused
Recent
disease
PLASMATIC
by many.
The
as Cofactors
of Aggregation
presented
VII,
here
IX,
and
and
indicate
that
(for
convenience,
X
platelet
membrane
and
in platelet-rich
plasma
for the
aggregation
K-dependent
prothrombin
coagulation
complex)
are cofactors
for the
by ADP,
epinephrine,
ofwashed
normal
platelets
are
ad-
aggregation
collagen,
by
bovine
of
and
fibrino-
gen.
MATERIALS
Aggregation
Broomall,
All
was
Pa.,
tests
were
plasma
(PRP)
sodium
citrate,
platelet
from
donors
carefully
Platelet
was
integrator
a sample
obtained
0.129
Aggregometer
and
using
M,
from
for
Model
(Linear
volume
I part
screened
METHODS
ofO.5
intake
of
Chrono-Log
Corp.,
Corp.),
Model
703.
ml.
centrifugation
to 9 parts
300,
Instrument
at
1700
of venous
aspirin
and
way:
PRP
rpm
blood)
for
7 mm
normal
other
drugs
of
human
known
fresh
blood
to
affect
aggregation.
Washed
for
the
a recorder
at 37’C
(3.8%
obtained
in
with
performed
Platelet-rich
citrated
measured
equipped
AND
platelets
10 mm,
suspended
the
(WP)
in saline
Epinephrine
were
sedimented
obtained
platelets
to a concentration
was
obtained
in the
washed
following
three
times
of 50,000-60,000
from
Parke-Davis
with
platelets/cu
adrenalin
was
0.85%
centrifuged
saline
at 4000
(0.1434
M)
rpm
and
re-
mm.
chloride
solution,
containing
1 mg
epinephrine/ml.
From
the Department
Submitted
A ddress
Miami
for
School
© 1975
Blood,
March
of Pathology.
1. /974;
reprint
requests:
of Medicine.
by Grune
Vol. 45, No.
accepted
& Stratton.
1 (January),
John
P.O.
Box
University
July
B.
520875,
of Miami
School
of Medicine.
Miami,
Fl. 33/46.
2. 1974.
Miale,
M.D.-
Biscayne
Department
Annex.
Miami,
of
Florida
Pathology,
University
of
33152.
Inc
1975
97
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98
MIALE
Adenosine
from
diphosphate
Schwarz-Mann.
added
to 0.5 ml of PRP
Collagen
pared
to
100 ml
more
of chilled
0.85%
rpm
adenosine-5’-diphosphate-disodium
bovine
and
in 0.85%
achilles
modified
saline
and
was centrifuged
at 4000
used
from
to Hovig,’3
suspension
was
solution
saline
was
(2
diluted
H2O),
as needed,
KENT
obtained
and
0.02
ml
was
or WP.
was obtained
according
The
mixed
at 4000
for
5 mm
at 4’C.
was
human,
tendon
in the
in a Waring
rpm
The
(Sigma
for
5 mm
milky
Chemical
following
Co.,
St.
way:
4 of
reagent
Blendor
for
5 mm
the
supernate
at 4’C
supernate
and
was
frozen
in
Louis,
Mo.),
powder
at
pre-
was
added
maximum
speed.
centrifuged
aliquots
at
once
105’F
-
and
as indicated.
Thrombin
and
used
diluted
with
the stock
with
used
equal
parts
was
was
mg/mI
of
N.J.)
30 mg/mI
1M
factors
II,
VII,
centration
XII,
showed
with
of eluate,
and
added).
IX,
and
of factor
IX.
no
The
of
Inc.,
days
after
and
frozen
The
There
test,
prothrombin
time
of about
the
plasma
and
of antiserum
serum)
and
using
used
for
A typical
inhibition
of
albino
of
bled
powder
sodium
citrate,
Piscataway,
M
II
VIII,
to
a 200-fold
factor
XI,
I ml
by
three
factor
the antigen
anti-IgM,
intramuscular
(Amphogel,
intravenously
cardiac
puncture
time
oxalated
plasma
is shown
for
the
diluted
15 mm
at
with
in-
Wyeth
at
10-day
and
the
Labora-
intervals.
serum
Ten
separated
37’C
powder
(0.1
then
adsorbed
with
on the plasma-antiserum
Table
I;
at dilutions
there
is very
M
sodium
BaSO4
1:10,
1:30,
Table 1. Inhibition of the Prothrombin Time of Normal
Anti-prothrombin
Complex Antiserum
Plasma by
0
Simplastin.
1:40,
0.2
41.5
0.2
328.4
0.1
1:20
0.2
86.6
0.1
1:30
0.2
59.5
0.1
1:40
0.2
51.9
0.1
1:50
0.2
plasma
diluted
with
tAntiserum
dilutions
made in dilute
plasma,
time determined
immediately
after the 15-mm
(General-Diagnostics,
Morris
normal
incubated
incubation
Plains,
N.J.).
for
15 mm
1:50.
the
pro-
Time
49.4
BaSO4-adsorbed
period.
9
of
The
and
of
Prothrombin
(sec)
human
at
room
plasma.
temperature
and
a
added,
0.1 g/ml
1:10
0.1
to
diluted
oxalate
prolongation
lhromboplastin-j
Calcium
(ml)
serially
powder,
using
1:20,
one-stage
plasma
antiserum
mixture
of
specificity.
of a modified
with
BaSO4
tested
immunologic
BaSO4-adsorbed
with
was
in
and
inhibition
oxalate,
performed
(R80)
activity
by determining
adsorbed
0.1
Simplastin
for
con-
or
antisera,
marked
human
acid-
assays
anti-IgG,
rabbits
coagulation-inhibition
assayed
Antiserumt
Concentration
citrated
boric
individual
specific
suspension
by
M
way:
BaSO4
(0.05
and
factor
factor
following
or anti-a-trypsin.
virgin
injection
0.2
anti-$-lipoprotein,
of Al(OH)3
were
of sodium
studies
study
Plasma’
(ml)
‘Normal
diluting
reconstituted
Chemicals,
buffer
ml,
V,
the
with
of
borate
against
antitransferrin,
one
normal
I part
these
with
protein/lOO
female
in
Fine
I g of
N.J.)
plasma
volume
Sephadex
antifibrinogen,
I ml
obtained
a small
factor
40 sec, incubated
a prothrombin
antiserum
the
Plains,
A-50(Pharmacia
demonstrable
both
was
previously
per
with
DEAE
no
rabbits
for
activity
time
parts
by
104’F.
-
prothrombin
with
reconstituted
prepared
ml.
oxalated
concentration
with
and
the
assayed
Coagulation-inhibition
was
human
a 40-fold
in young
Pa.)
at
were
fresh
averaged
with
mixed
injection
Morris
By immunoelectrophoresis
prepared
in aliquots
antisera
N.J.),
were
of 150 mg/lOO
from
was
lines
of antigen
last
Raritan,
solutions
Diagnostics,
BaSO4
from
antigen
ranged
Philadelphia,
the
the
Sephadex
anticeruloplasmin,
was
I ml
Co.,
Working
II-VII-IX-X)
of
elution
activity.
precipitin
antiserum
jections
tories,
X
(anti
from
dry
The
antihaptoglobin,
,
U/mI.
to a concentration
adsorption
elution
nor any thrombin
anti-C3
(General
by
eluate
NaCI
fibrinogen
saline
antiserum
prepared
ofthe
NaOH,
and
plasma),
readsorption
Pharmaceutical
7.5
bovine
complex
antigen
(Ortho
to contain
saline.
ofH2O
Antiprothrombin
The
Fibrindex
0.85#{176}csaline
solution
Fibrinogen
with
(50
(ADP)
A 4.57-mM
AND
the
clotting
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PLATELET
99
AGGREGAIION
Table
2. Inhibition of Assays
of Prothrombin
Factors by Anti-prothrombin
Complex
Complex
Antiserum’
Normal
Normal
‘Normal
serum,
citrated
0.02
thrombin
1:50.
1:10 dilution
also
strongly
mal
human
could
Factor
IX
>100%
1.1%
Factor
X
100%
0.5%
1:10
100%
diluted
at room
1:10
was
and
used
to
with
the antiserum
significant
batches
and
antiserum
by immunoelectrophoresis
be easily
removed
by adsorption
then
assayed
the
assays
for
for
antiprothrombmn
complex
anti-
of
a faint
onto
an excess
1
2
3
4.
at
the
factors
each
antiserum
dilution
II,
saline
factors
an
partial
concentration
by 50#{176},.The
VII,
was
IX,
and
incubated
II, VII,
IX,
of
factors,
the
of
thromboplastin
and
antiserum
X (Table
for
X.
time.
but
2). Nor-
15 mm
As
shown
the
A
was
at
room
in Table
specificity
of
is questionable.
gel diffusion,
Fig. 1. Composite
tracings, aggregation
and inhibition
of aggregation
in
PRP.
(A)
Controls:
(1)
collagen.
of
and
4
B
evident
showed
A
epinephrine,
(5)
prothrombin
complex
antiserum.
(B)
Inhibition
of
aggregation,
preincubation
with
R80
antiserum
followed
by (1) thrombin,
(2)
epinephrine,
(3)
ADP, (4)
is still
buffered
antibody
3
(2) collaADP, (4)
R80 anti-
plus
ml,
inhibition
systems
with
2
thrombin alone,
gen alone, (3)
0.1
by 80#{176},,a 1:20
1:10
in
of polyvalent
of
effect
assay
diluted
reduction
in the presence
the
the PTT
in individual
plasma
saline,
for 15 mm.
determine
prolonged
42%
1.2%
buffered
with
temperature
antiserum,
system
inhibitory
Occasional
fibrinogen
>100%
citrated
was
the assays
VII
of antiserum
temperature
2, there
II
Factor
plasma
by
similar
Plasma
+ Antiserum
Factor
ml, incubated
time
A
Plasma
precipitin
but
of human
line
this
when
minor
fibrinogen.
tested
antifibrinogen
against
human
component
lmmunoelectrophoretic
From www.bloodjournal.org by guest on July 31, 2017. For personal use only.
MIALE
100
analysis
of
precipitin
the
antiserum
lines,
ofthe
but
prothrombin
Normal
bino
serum,
Mo.)
and
the
be made
control,
with
antisera
antihuman
City,
for
It was adsorbed
Monospecific
Calif),
against
can
antigen
at this
used
time
for
that
injection
these
showed
correspond
to
four
the
KENT
distinct
four
factors
complex.
rabbit
rabbit.
(R80)
no claim
AND
used
IgG
were
(Hyland),
antihuman
was
BaSO4
obtained
prior
the
following:
antihuman
complement
by cardiac
puncture
to use, as with
antihuman
a-l
lipoprotein
from
a normal
donor
al-
the antiserum.
fibrinogen
(Hoechst
(Hyland,
Costa
Mesa,
Co.,
Kansas
Pharmaceutical
C3 (Hyland).
RESULTS
Aggregation
and Inhibition
As shown
(final
in Fig.
1A, normal
concentration
collagen
(1:2
U/ml.
3.7
dilution
aggregating
and
epinephrine
sera
was
were
it was
used
serum,
As shown
collagen
0.04
(150
(1:2
dilution
ml of R80. They
mg/100
ml) were
When
a suspension
and
ADP,
bovine
rabbit
plement
C3, or antihuman
Studies
serum,
on the Specificity
A series
of absorption
to
for
3 mm
R80
the
No
inhibition
the
antiprothrombin
0.3
PRP
no
IgG,
by
response
to ADP
when
the
and
other
antiserum:
normal
antihuman
washed
platelets
M),
l0
of Washed
complement
of stock),
were
epinephrine
not
thrombin
(final
platelets
fibrinogen
was
(curve
when
and
aggregated
(final
0.3
incubated
with
8) there
was
the addition
epinephrine,
fibrinogen,
alpha-I
ADP
0.2
0.25 ml of bovine
platelet
suspension.
bovine
and thrombin.
No inhibition
in place of the antiprothrombin
antihuman
by
concentration
concentration
aggregated
normally
when
added
to 0.25 ml ofwashed
of washed
Platelets
U/mi),
or
fibrinogen
antiserum
fibrinogen
could
antihuman
(final
mg/mI),
R80
complete
inhibition
was bovine
fibrinoand
colla-
be demonstrated
complex
antiserum:
IgG,
antihuman
com-
lipoprotein.
of the Antiserum
studies
was
done
to establish
normal
of washed
that
the
g and
aggregation
inhibition
effect
noted
human
platelets
platelets,
final
concentration
140,000/cu
mm and 50,000/cu
mm,
respectively)
room
temperature
and 4 hr at 4#{176}C.
The adsorbed
antiserum
was
at 3000
followed
epinephrine,
complex
specific:
1. The antiserum
was adsorbed
with washed
of antiserum
+ 0.1 or 0.04
ml of a suspension
centrifugation
produced
be demonstrated
antihuman
mg/mI),
concentration
by ADP,
secondary
could
ofAggregation
gen, or bovine
fibrinogen
when other
sera were used
Normal
(R80)
2B) aggregation
and
of
(final
ml
(Fig. 2B) followed
by bovine
fibrinogen
of aggregation.
Inhibition
also occurred
gen
alone
by ADP
0.2
lipoprotein.
2A,
x
thrombin
(Fig.
fibrinogen,
Inhibition
1.84
and
produced
concentration
0.04
inhibited,
of
alpha-l
in Fig.
concentration
was
in place
and
that
eliminated.
C3, or antihuman
Aggregation
with
were
(final
of antiserum
found
antihuman
ofaggregation
solution),
addition
thrombin
was
patterns
incubated
agent
collagen,
rabbit
the
in PRP
M), epinephrine
x l0
of stock
In contrast,
aggregation.
When
PRP
ofAggregation
determined
was
(0.5 ml
platelet
for 30 mm at
harvested
after
as
described
From www.bloodjournal.org by guest on July 31, 2017. For personal use only.
PLATELET
AGGREGATION
101
A
B
R 80
Composite
tracings,
aggregation,
and inhibition
of aggregation
of washed
platelets.
(1) bovine fibrinogen added,
(2) ADP, (3) thrombin,
(4) epinephrine,
(5) collagen,
(6) R80 anti-prothrombin
complex
antiserum.
(B) Inhibition of aggregation:
(1) ADP plus fibrinogen, (2) epinephrin
plus flbrinogen, (3) fibrinogen alone, (4) collagen plus fibringen, (5) thrombin
plus flbrinogen,
(6-10)
preincubation
with
antiserum
followed
by: (6) collagen and fibrinogen,
(7) ADP plus flbrinogen, (8) fibrinogen alone, (9) epinephrine
plus flbnnogen,
(10) thrombin
plus
fibrinogen.
FIg. 2.
(A) Controls:
2
FIg.
3.
Typical
tracings
(composite)
of
the effect
of adsorbed
antisera on platelet
aggregation
in PRP by epinephnne.
(1)
Antiserum adsorbed
with human platelets,
(2)
antiserum
adsorbed
(BaSO4 -eluate),
with
BaSO4-adsorbed
untreated
antiserum.
(3)
with
antiserum
human
antigen
adsorbed
plasma,
(4)
4
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102
MIALE
above.
As shown
inhibiting
lets contain
It should
[Fig.
in Fig.
activity.
specific
be noted
3(1),
platelet-adsorbed
antiserum
loses
This is interpreted
receptor
sites for
as supporting
the
the antiprothrombin
that
alone
the
antiserum
has
AND
KENT
its aggregation-
concept
that the platecomplex
antibodies.
no platelet-aggregating
activity
lA,(5)].
2. The antiserum
prothrombin
time
plasma)
for 30 mm
serum
was
determined
>
was adsorbed
with BaSO4-adsorbed
600 sec (0.5 ml of antiserum
at room
temperature
and 4 hr
harvested
after
centrifugation
as described
above.
As shown
BaSO4-adsorbed
plasma
retains
at 3000
in Fig.
its original
normal
human
plasma,
0.1 ml BaSO4-adsorbed
at 4#{176}C.
The adsorbed
anti+
g and aggregation-inhibition
3(3) antiserum
adsorbed
capacity
to inhibit
platelet
with
aggrega-
tion. This is interpreted
as evidence
(BaSO4
eluate)
are active
in platelet
adsorbed
plasma
is rich in fibrinogen,
that only antibodies
present
in the antigen
inhibition.
Furthermore,
since the BaSO4there
is additional
confirmation
that the
inhibition
of antifibrinogen.
is not
due
to the
presence
3. The
the eluate
antiserum
was adsorbed
with a freshly
prepared
antigen
of BaSO4-adsorbed
normal
plasma
(0.5 ml of antiserum
antigen),
for
30 mm
serum
was harvested
termined
as described
to inhibit
platelet
the inhibition
prothrombin-complex
at room
temperature
4 hr at 4#{176}C.
The
adsorbed
anti-
by centrifugation
at 3000 g and aggregation
inhibition
deabove.
As shown
in Fig. 3(2), this antiserum
is unable
aggregation.
of
and
preparation,
+ 0. 1 ml of
platelet
This
is interpreted
aggregation
proteins.
as supporting
is specifically
due
the
to
concept
antibodies
that
against
DISCUSSION
Platelet
been
has
aggregation,
the subject
of
so intimately
involved
in hemostasis
and
thrombosis,
intensive
study.
Although
these
studies
have
partially
clarified
the classification
fect in von Willebrand’s
when
aggregation
takes
platelets
sidered
of qualitative
platelet
disorders
and
disease,
the physicochemical
reactions
place
are still nebulous.
The concept
aggregate
because
too simplistic
but
observations
that
normal
a “sticky”
fibrin
forms
may be in fact the last
platelets
ADP,
epinephrine,
collagen,
have
intrigued
investigators
mechanisms
responsible
for
are
the
platelet
dewhich
occur
of Apitz’4
that
on the surface
has been conof a series
of reactions.
Later
aggregated
by such
diverse
substances
as
thrombin,
and
ristocetin,
among
many
others,
and stimulated
much
research
into the definitive
the aggregation
reaction.
Most
investigators
feel
that ADP
is the final common
pathway
agents,
but there
is no agreement
on
the final reaction’5’7
for the aggregation
produced
the molecular
mechanisms
that
by various
precede
One area of special
interest
has been
the role of plasmatic
cofactors
in the
aggregation
reaction.’8
The concept
that a “plasmatic
atmosphere”
of proteins
is a requirement
for normal
platelet
function
was first proposed
by Roskam
in
l923.’
There
followed
several
reports2022
dealing
with the identification
of the
proteins
involved
calcium,23
two
and
VIII.
The
factor
definition
in platelet
coagulation
of fibrinogen
aggregation
proteins
are
as a cofactor
and
release
known
to
was
reactions.
act
derived
In
as cofactors,
from
in vitro
addition
fibrinogen
studies58
to
From www.bloodjournal.org by guest on July 31, 2017. For personal use only.
PLATELET
and
AGGREGATION
from
the
observations
ADP-induced
own
103
afibrinogenic
it has
plasma
been
in clinical
aggregation
centrations
of ADP
high.2
It is noteworthy
and
that
platelet
usually
but
aggregate
that
only
pointed
out
cases
of congenital
afibrinogenemia
is diminished.’4
Platelets
show
aggregation
diminished
suspended
normally
if the concentration
minute
amounts
of fibrinogen
that
not
infrequently
in vivo
and
Willebrand’s
in vitro,
it will
disease’#{176}” unless
molecule
which
ristocetin
is now
has
cofactor
in platelet
is factor
XII.’8’26
Attempts
to
identified
von
Willebrand
known,
but
activity,25
the
aggregation
identify
role
seems
well
aggregation
deficiencies
von
The
Willebrand
established.
cofactors
of coagulation
from
portion
is added.’2
of the
factors
are
the
not
use
been
subjects
of the
with
factor
mode
of action
factor
as a second
third
of
same
of the
or addition
criticism
additives.
mosphere”
vironment
We
necessary
around
constituents
“sponge”
of specific
began
are
with
the
for platelet
the platelet
onto
the
is not new.27
coagulation
have
factors
of the
working
XI
as yet not
hypothesis
plasmas
been
subject
the
of
aggregation
Furthermore,
the antiserum
we show that
purity
“plasmatic
at-
immediate
enof plasmatic
the
platelet
as
such as serotonin,28
towards
techniques
offers
methods
of coagulation
and
graphic
many
are
advantages
more
specific
proteins.
The
measurement
of the
cell membrane.
of platelets
in PRP
by ADP,
the inhibition
is not related
We
assume
proteins
a
adsorb
coagulaXII.3”38
Other
coagulation
reaction
of
the complex
epinephrine,
thrombin,
and
to an interaction
between
and the normal
plasma
in which
the platelets
are suspended,
the antiserum
inhibits
also the aggregation
of washed
platelets
fibrinogen.
directed
abthe
to the
of the
data indicate
that one or several
of the vitamin
K-dependent
in the prothrombin
complex
are required
for the aggregation
platelets.
An antiserum
prepared
against
the proteins
of
inhibits
collagen.
bovine
provides
at the
having
implicated.
by immunologic
a sensitive
Our
factors
normal
that
and myxovirus,3#{176} platelets
and
XII,35’36
and
possibly
use of platelet
aggregation
are
difficulty
of
It seems
a complete
relying
on
is not primarily
the
it is the adsorption
Immunologic
are the assays
occurring
the
hypothesis
over chemical
or indirect
approaches.
and many
times
more
precise
than
reactions
means
plus
platelet
membrane.
The concept
In addition
to adsorbed
substances
plasminogen,29
VIII,33
IX,
test
by chemical
concerned
aggregation
but rather
immunoglobulins,29
tion factors
V,3”32
The
factors
as far as substrates
von
VIII
cofactor
rewarding.’8
probable
that even in the most
severe
deficiencies
there
is not
sence of the implicated
coagulation
factor.
In vitro
experiments
subtraction
is
it causes
platenormal
platelets
A possible
by
have
con-
ADP
required,
afibrinogenemic
in fact contain
small amounts
of fibrinogen.3
is an antibiotic
no longer
used for therapy
because
and thrombocytopenia.24
While
it aggregates
platelets
or that
low
of
so-called
plasma
does
Ristocetin
let agglutination
not aggregate
factor
VIII,
in their
with
of
that
the
action
the
prothrombin
support
the
of the
antibodies
complex
at
in the
the
platelet
for
by
serum
mem-
brane.
Data
effect
on
thrombin
are
presented
which
the antiserum
used
complex
proteins:
opinion
that
is specifically
attributable
(1) there
is no aggregation
the
aggregating-inhibiting
to antibodies
inhibition
by
to pronormal
is
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104
MIALE
rabbit
serum,
human
antihuman
complement
(2) adsorption
fibrinogen
C3 antiserum,
of
the
antiserum
aggregation-inhibition
(BaSO4
eluate)
and
of the antiserum
(4) adsorption
the
antihuman
IgG
alpha-i
lipoprotein
with
does not alter
its
serum
with antigen
its aggregation-inhibition
antiserum,
or antihuman
normal
human
antiserum,
normal
anti-
BaSO4-adsorbed
human
plasma
of
platelets
the antiactivity;
also
abolishes
activity.
There
is a theoretical
possibility
antiserum
is due to an antibody
that
the aggregation-inhibiting
to an unidentified
protein
activity
of
in the antigen,
but we feel this to be unlikely.
First,
we show
by immunoelectrophoresis
least some
of the possible
contaminating
proteins
are not demonstrable
antigen.
Second,
the antiserum
alone does not aggregate
platelets,
and
gests,
at least,
that no specific
antiplatelet
antibodies
it is possible
that as yet unidentified
proteins
may
eluted
with
altered
citrate,
the
by adsorption
strably
powerful
aggregation-inhibition.
absolute
powerful
with
Finally,
of
the
clot-inhibiting
specificity
of
are in progress
to
one, several,
or all four
powerful
inhibition
the II-VII-X
plastin
time
strabie
antifactor
antifibrinogen,
V; while
aggregation,
The
or
in
of the
group,
implicates
the
they
have
the
platelet
sequence
time
XI,
may
excluded
in the
plus
the role of platelets
of the coagulation
to the
improbable
setting
by
only
the
XII,
cofactors
demon-
be
proved
Work
by
is now
in
is directed
complex.
The
antiserum
of
used
implicates
the partial
contains
no
thrombodemon-
anti-factor
involved
in
VIII,
or
platelet
study.
prothrombin
other
complex
coagulation
of fibrin
reactions
coagulation
are
are
factors
in hemostasis.
proteins
and the
formation
that the
the entire
can
not
the
the ones active
in
ofthe
above
offer
the specificity
prothrombin
test
anti-factor
also be
antiserum,
factors.
inhibition
antiserum
in this
factors
membrane,
leading
brane.
It is not
in a microcosmic
This
that at
in the
this sug-
Third,
while
BaSO4
and
that
are
none
coagulation
less striking
IX-X.
The
anti-factor
of these
that
plicated,
expands
many,
if not all,
the
prothrombin
been
demonstration
reaction.
pure
by
of the
suggests
determine
whether
ofthe
factors
in the
while
the
factors
some
plasma,
to the prothrombin
complex
it must be admitted
that
using antisera
prepared
to acceptably
progress
toward
this goal.
Studies
towards
are present.
adsorbed
be
activity
BaSO4-adsorbed
antibodies
proof
KENT
antiserum;
activity;
(3) adsorption
abolishes
its aggregation-inhibition
with
AND
It can be
phospholipid
present
on
at the platelet
sequence.
present
on
previously
im-
postulated
involved
the
platelet
membrane
Aggregation
that
in
mem-
replicate
may be
both
the manifestation
of the coagulation
sequence
and the reaction
by which
a self-generating
sequence
is maintained.
The evidence
at present
suggests
that
there
is an interaction
between
the aggregating
agent
and coagulation
proteins
adsorbed
onto
the platelet.
ing to release
of ADP
and
sumed
that
activation
prothrombin
to fibrin.
The interaction
other
intracellular
of the
to thrombin
Aggregation
coagulation
which
in turn
is an expression
alters
the platelet
constituents.
sequence
converts
of these
leads
membrane,
It can also
to
the
fibrinogen
at
leadbe as-
conversion
the
of
membrane
reactions.
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and
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viscous
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J: Aggrega-
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1975 45: 97-106
Prothrombin complex proteins as cofactors in platelet aggregation. I.
Inhibition of aggregation by antiserum
JB Miale and JW Kent
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