PATIENT
WITH INHIBITOR AGAINST F-IX.
PE
Makris,
Poumbouridou
E,
Constantinidis
C,
Pithara
E.
Thrombosis
& Haemostasis Unit,
A'
Medical Propedeutic Clinic
University
of Thessaloniki-
Macedonia-
GREECE
introduction
It
is widely stated that about 10% of all haemophilia patients develop antibodies
against coagulation factors given as replacement therapy during bleedingepisodes.
This
type of reaction is more rarely seen in Haemophilia B.
It
should be pointed out that the titre of the inhibitor is a necessary information
either for the treatment of acute situations or the preparation of operations.
A
great number of sophisticated and expensive strategies have been designed
to decrease the titre of the inhibitor such as :
a)
High doses of FIX, enough to neutralise FIX inhibitor and increase FIX
levels up to 40% - 80%, an approach extremely expensive.
b)
Plasmapheresis.
c)
Immune suppression, of inhibitor's production.
d)
Use of agents capable to activate other coagulation pathways, regardless
of FVIII or FIX.
Our
case.
We
described a 64 year-old haemophiliac, FIX 9%, with inhibitor titre of 1.5
Oxford units, and occlusion of three coronary arteries, who successfully
underwent coronary by-pass.
Haemophilia
B as well as the existence of FIX inhibitor were diagnosed because of an
uncontrolledbleeding during the
cardiac catheterization. Patientreferred
minor haemorrhagic manifestations since his childhood.
Operation
was performed after successful suppression of inhibitor's production using
a five-days regimen of iv cyclophosphamide after incubation with concentrated
monoclonal FIX.
During
the operation and until the third postoperative day the patient was given
FIX 100 unit/Kg BW/8h.No rebound
phenomenon was detected during his three month's follow up.
CORONARY
ARTERIES BY PASS IN A B-HEMOPHILIAC WITH INHIBITOR AGAINST F-IX. PE
Makris, Poumbouridou E,
Constantinidis
C, Pithara E.. Thromb
Haem Unit, A' Med. Prop. Clinic Univ of Thessaloniki- Greece.
It
is widely stated that about 10% of all hemophilia patients develop antibodies
against coagulation factors given as replacement therapy during bleeding
episodes. The molecular mechanisms found responsible in hemophilia B patients
are highly heterogeneous. Among them, point mutations are responsible for
about 95% of all disorders. The development of FIX inhibitor in no- haemophiliacs
is extremely rare, and is mainly reported in multi-parum women and in patients
with auto immune diseases. It should be stressed that the titre of the
inhibitor is a necessary information either for the treatment of acute
situations and especially for the organisation of operations. A great number
of sophisticated and expensive strategies have been designed to decrease
the titre of the inhibitor such as a) High doses of FIX ,enough to neutralise
FIXinhibitor and increase FIX level
up to 40% to 80%,an approach extremely expensive. b) Plasmapheresis. c)
Immune suppression, of inhibitor' s production.d)
Use ofagents capable to activate
the final common pathway of thecoagulation
cascade, independently of FVIIIor
FIX .
Our
case. We described a 64 year-old haemophiliac, FIX 9%, with inhibitor,
titre 1,5 Oxford units, and occlusion of three coronary arteries, who successfully
underwent coronary by-pass. The diagnosis of hemophilia B and FIX inhibitor
was made because of an uncontrolledbleeding
during the cardiac catheteriazation. He refereed minor hemorrhagic manifestations
since his childhood. The operation was performed after successful suppression
of inhibitor' s production using a five-days regimen of i.v cyclophosphamide
after incubation with concentrated monoclonal FIX. During the operation
and until the third postoperative day the patient was given FIX 100 unit/KgBW/8h.No
rebound phenomenon was detected in his three month's follow up.
MULTI-POTENTIAL
ANTIBODIES AGAINST ACTIVATED PLATELETS ACTIVATION THROUGH Gp-Ib.
Makris
S, Poumbouridou E,Gerotziafas
G, Constantinidis C, Pithara E, PE Makris. Thromb Haem Unit, A' Med
Prop Clinic of the University of Thessaloniki-Macedonia-GREECE.
The
production in rabbits of multi-potential antibodies against activated human
platelets which stimulate aggregation (MaK-1) led us to try to define their
function and molecular structure. It is well known that monoclonal inhibition
or stimulation antibodies could be revealed in vitro. The stimulating antibodies
act by binding with low molecular weight protein, molecules such as CD9,
and induce platelet aggregation, either by activation of the immune response
receptors, FcãRII,
or by complement’s activation. These pathways can be checkedusing
a) special monoclonal antibodis (Mo-a) the Mo-a IV.3, for the FcãRII
receptor, which inhibit aggregation. b) special Mo-a (Leupeptin) which
inactivate the complement, c) having in mind that the functional activation
of platelets is based on glycoproteins Gp-Ib-IX, IIb-IIIa, Ia-IIa, Ic-IIa,
we used special Mo-abs to trigger or block their activation.
Materials
and methods;
Mo-abs mouse anti GpIIb-IIIa, GpIb, GpIIIa from DAKO co. Mo-ab IV.3 and
Mo-ab Pl2-49 were kindly offered by Dr Lecompte. We checked the functional
properties of multipotential Mak-1 in PRP and using the computerised aggregometre
platelet ionised calcium Chrono-Log co.
Results;
Platelet aggregation; a)Incubation
of Mo-a IV.3 for three minutes in PRP can not inhibit plts aggregation
triggered by Mak-1 abs.b) The addition
ofPl 2-49 did not modify platelet
aggregation induced by Mak-1 abs. c) Addition and incubation, for three
minutes, of Mo-ab GpIIb-IIIa did not inhibit aggregation by Mak-1. We have
the same results with Mo-ab GpIIIa. d) Platelet response was reduced with
Mo-a GpIb addition.ATP secretion;
We detected ATP release in all experiments resulting in platelet aggregation.
The results lead us to conclude that the activation of platelets may be
mediated through complement and Gp Ib-IX complex.
MULTI-POTENTIAL
ANTIBODIES AGAINST
ACTIVATEDPLATELETS
: I-ACTIVATION THROUGH Gp-Ib.
Makris
S, Poumbouridou E,Gerotziafas
G,
Constantinidis
C, Pithara E, PE Makris..
Thrombosis
& Haemostasis Unit,
A'
Medical Propedeutic Clinic
University
of Thessaloniki-
Macedonia-
GREECE
introduction
The
production in rabbits of multi-potential antibodies against activated human
platelets which stimulate aggregation (MaK-1) led us to try to define their
function and molecular structure.
It
is well known that
monoclonal
inhibition
or
stimulation antibodies
could
be revealed in vitro.
Stimulation
antibodies act
by
binding with low molecular weight protein, such as CD9, and induce platelet
aggregation,
either
by activation of the immune response receptors, FcãRII,
or
by complement’s activation.
These
pathways can be checkedusing :
a)
special monoclonal antibodies (Mo-a) the Mo-a IV.3, for the FcãRII
receptor, which inhibit aggregation.
b)
Mo-a (Leupeptin) which inactivate the complement,
c)
having in mind that
functional
activation of platelets is based on glycoproteins
GpIb-IX,
IIb-IIIa, Ia-IIa, Ic-IIa, we used special Mo-abs to trigger or block their
activation.
material
and methods
Mo-abs
anti GpIIb-IIIa, GpIb, GpIIIa from DAKO Co.
Mo-ab
IV.3 and Mo-ab Pl2-49 were kindly offered by
Dr
Lecompte.
We
checked the functional properties of multipotential Mak-1 in PRP usingcomputerised
aggregometre PICA Chrono-Log Co.
results
Platelet
aggregation :
a)
Incubation of Mo-a IV.3 for 3 min in PRP can not inhibit plts aggregation
triggered by Mak-1 abs.
b)
The addition ofPl 2-49 did not modify
plts aggregation induced by Mak-1 abs.
c)
Addition and incubation, for 3 min, of Mo-ab GpIIb-IIIa did not inhibit
aggregation by Mak-1.
We
have the same results with Mo-ab GpIIIa.
d)
Platelet response was reduced with Mo-a GpIb addition.
ATP
secretion:
We
detected ATP release in all experiments resulting in plts aggregation.
conclusion
These
results led us to conclude that the activation of platelets may be mediated
through complement and
Gp
Ib-IX complex.