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149 Thrombin generation, endothelial cell function and fibrinolytic potential in homozygous sickle cell disease and beta-thalassemia
POSTER P R E S E N T A T I O N S : Patophysiological studies !
53
m
147 Heterogeneous distribution of the subclass IgG anti-streptokinase antibodies *REGNA ULT V, */**LECOMPTE T, and **DANCH1N N on behalf of the French Clinical investigators (Besanfon, Papeete, Paris) *UMR CNRS 7563, Facultd de Mddeeine et **CHU, Nancy, France Serum concentration o f IgG antibodies to streptokinase (SK) was measured by ELISA in three groups of patients: 95 with coronary artery disease (group I), 21 in the 2 "d year after treatment with SK (group II) and 31 from French Polynesia (group III) in order to characterize anti-SK antibodies secondary to SK-therapy and/or to streptococcal infection. Since parallel concentration response curves were observed with 137 out o f 147 sera, specific anti-SK IgG from one serum was immunopurified on immobilized SK using the surface plasmon resonance technology, quantified with an ELISA for total IgG and used as standard in the anti-SK ELISA. Concentrations in specific anti-SK l g G > 100 pg/ml were found in all sera from group II and in 15 and 12 sera from groups I and III respectively. A correlation was observed between the inhibition o f the clot-lytic effect o f SK and the concentration of anti-SK IgG. IgG subclass distribution was determined by ELISA m
for these 48 sera. T h e optimal conditions for each assay with anti-IgG subclass antibodies and the lack of cross-reactivity a m o n g these antibodies were determined in initial experiments using m y e l o m a proteins. Specific anti-SK IgG from the four subclasses were found in various combinations, lgG2 were detected in 24 sera, 6 from group I, 7 from group II and I 1 from group III. These results are at variance with previous reports suggesting that antibodies to SK are only IgG1. The relative inefficiency of macrophage clearance o f IgG2-containing imm u n e complexes might favor platelet activation. One bi-allelic polymorphism o f the platelet Fc),RIIa modulates the binding of h u m a n IgG2. Additional studies are needed to define the relations between IgG subclass-specific anti-SK antibodies, FcyRIIa polymorphism and SK-induced antibody-mediated platelet aggregation.
III
148 Disseminated intravascular coagulation, fibrinolysis-and proteolysis-syndrome (DIC-F-PS) in two patients (PTS) with purulent cholangitis or portal sepsis, complicated with acute hepatic failure (AHF). Benefit from therapy with plasma derivates (PD) (AT IIIc, FFP) KULLMER ,I, LERCH L, KRON1GER A. SEITZ R, SCHUERMANN M, and EGBRING R Department of Hematology/Oncology/lmmunology, Philipps-University, 35033 Marburg, Germany Background. Besidesviral hepatitis,CCL4_and ammanitapalloidesintoxication,even pu-
rulentcholangitisor portal sepsisare sometimescomplicatedwithDIC-F-P-Sand withAHF.
vated at the beginning of the severe illness in both pts. The lowest or the highest values respectivelyduring the first week are shown below. Diagnosis of Quick pt. % Purulent cholan~itis
22
ATIII
Plsg.
TAT
PAP
ELPctlA 4F~B~30ng/ml
picomol/I
2424 FI+2 ng/ml 3.3
%
%
ng/ml
ng/ml
25
22
15.5
n.d.
676
48
5
92
3585
418
Aim of the study. We now report about AHF in one patient with purulentcholangitisand
one patientwith portal sepsis.Histologicalexamination(HE) of liver biopsyconfirmedthe purulentcholangitisin pt I andportal sepsisin pt2. Liver cell necrosesare foundin bothHEs. Methods. Coagulationfactors and inhibitorshad been determind by routine laboratory. Moreoverproteinaseinhibitorcomplexes(PIC) such as Thrombin-Antithrombin-111(TAT), Plasmin-a-2-Antiplasmin (PAP), Elastase-~-l-Antitrypsin (ELP-ct-I-AT) and Prothrombin-fragmentF I+2 (FI +2) had been determinedas DIC-Factivationmarkersor of polymorphonuclearleucocytes(PMNL's) activationand ELP releaseinto the circulation. Results. Duringacute phases, before substitutionwith PDs some coagulationfactors and inhibitorsare severelydecreased.The o.m. PICs and FpB~30-43are foundto be highlyele-
Portal sepsis 7
Conclusion. Intensive care and PDs substitution(ATIIIc, ptl=27.000 IU, pt=30.000 IU
and differentamountsof fresh frozenplasma)improvedthe severecoagulationdefectsand stop the inhibitorconsumption.The parameterstypicallyelevatedin DIC-F-P-Sdecreased to normal values together with recovery. Both pts suffered transientlyfrom prltcoma or coma hepaticumrespectivelybut survived after long term intensivecare.
l
149 Thrombin generation, endothelial cell function and fibrinolytic potential in homozygous sickle cell disease and betathalassemia *GOUBRAN F, *NEVENE RAMSS1S, and **ATEF A *Clinical Pathology and **Pediatric Dep. Suez Canal University, Egypt Several studies have indicated that vaso-occlusion in sickle cell disease (SCD) is a multi-factorial process rather than a simple obstruction by sickled cells. Red blood cells from patients with h o m o z y g o u s betathalassemia behaves as procoagulant cells. W e tried to study thrombin generation and endothelial cell markers in relation to the fibrinolytic potential in SCD patients (n=9) during crisis and in the steady state as well as in h o m o z y g o u s beta-thalassemia patients (n= 11). The obtained results were compared to those o f age matched controls. Thrombin/antithrombin III complex (TAT), prothrombin fragment 1+2 (Fl+2) and thrombomodulin (TM) levels, as assessed by commercial E L I S A ' s , were mostly elevated in SCD during crisis followed by SCD in quiescent states and thalassemics when compared to controls. The activity of tissue type plasminogen activator (t-PA), plasminogen activator in-
hibitor type 1 (PAl-l), o~2-antiplasmin and histidine rich glycoprotein (HRG), determined by chromogenic substrate assays, were slightly increased during crisis in S C D and were not changed from control levels in the quiescent states and thalassemic patients. The amplitude o f thrombin generation was not asscociated with similar elevation in the fibrinolytic activity in SCD during crisis denoting that defective fibrinolysis participates in the vast-occlusive complication of SCD.
53
m
147 Heterogeneous distribution of the subclass IgG anti-streptokinase antibodies *REGNA ULT V, */**LECOMPTE T, and **DANCH1N N on behalf of the French Clinical investigators (Besanfon, Papeete, Paris) *UMR CNRS 7563, Facultd de Mddeeine et **CHU, Nancy, France Serum concentration o f IgG antibodies to streptokinase (SK) was measured by ELISA in three groups of patients: 95 with coronary artery disease (group I), 21 in the 2 "d year after treatment with SK (group II) and 31 from French Polynesia (group III) in order to characterize anti-SK antibodies secondary to SK-therapy and/or to streptococcal infection. Since parallel concentration response curves were observed with 137 out o f 147 sera, specific anti-SK IgG from one serum was immunopurified on immobilized SK using the surface plasmon resonance technology, quantified with an ELISA for total IgG and used as standard in the anti-SK ELISA. Concentrations in specific anti-SK l g G > 100 pg/ml were found in all sera from group II and in 15 and 12 sera from groups I and III respectively. A correlation was observed between the inhibition o f the clot-lytic effect o f SK and the concentration of anti-SK IgG. IgG subclass distribution was determined by ELISA m
for these 48 sera. T h e optimal conditions for each assay with anti-IgG subclass antibodies and the lack of cross-reactivity a m o n g these antibodies were determined in initial experiments using m y e l o m a proteins. Specific anti-SK IgG from the four subclasses were found in various combinations, lgG2 were detected in 24 sera, 6 from group I, 7 from group II and I 1 from group III. These results are at variance with previous reports suggesting that antibodies to SK are only IgG1. The relative inefficiency of macrophage clearance o f IgG2-containing imm u n e complexes might favor platelet activation. One bi-allelic polymorphism o f the platelet Fc),RIIa modulates the binding of h u m a n IgG2. Additional studies are needed to define the relations between IgG subclass-specific anti-SK antibodies, FcyRIIa polymorphism and SK-induced antibody-mediated platelet aggregation.
III
148 Disseminated intravascular coagulation, fibrinolysis-and proteolysis-syndrome (DIC-F-PS) in two patients (PTS) with purulent cholangitis or portal sepsis, complicated with acute hepatic failure (AHF). Benefit from therapy with plasma derivates (PD) (AT IIIc, FFP) KULLMER ,I, LERCH L, KRON1GER A. SEITZ R, SCHUERMANN M, and EGBRING R Department of Hematology/Oncology/lmmunology, Philipps-University, 35033 Marburg, Germany Background. Besidesviral hepatitis,CCL4_and ammanitapalloidesintoxication,even pu-
rulentcholangitisor portal sepsisare sometimescomplicatedwithDIC-F-P-Sand withAHF.
vated at the beginning of the severe illness in both pts. The lowest or the highest values respectivelyduring the first week are shown below. Diagnosis of Quick pt. % Purulent cholan~itis
22
ATIII
Plsg.
TAT
PAP
ELPctlA 4F~B~30ng/ml
picomol/I
2424 FI+2 ng/ml 3.3
%
%
ng/ml
ng/ml
25
22
15.5
n.d.
676
48
5
92
3585
418
Aim of the study. We now report about AHF in one patient with purulentcholangitisand
one patientwith portal sepsis.Histologicalexamination(HE) of liver biopsyconfirmedthe purulentcholangitisin pt I andportal sepsisin pt2. Liver cell necrosesare foundin bothHEs. Methods. Coagulationfactors and inhibitorshad been determind by routine laboratory. Moreoverproteinaseinhibitorcomplexes(PIC) such as Thrombin-Antithrombin-111(TAT), Plasmin-a-2-Antiplasmin (PAP), Elastase-~-l-Antitrypsin (ELP-ct-I-AT) and Prothrombin-fragmentF I+2 (FI +2) had been determinedas DIC-Factivationmarkersor of polymorphonuclearleucocytes(PMNL's) activationand ELP releaseinto the circulation. Results. Duringacute phases, before substitutionwith PDs some coagulationfactors and inhibitorsare severelydecreased.The o.m. PICs and FpB~30-43are foundto be highlyele-
Portal sepsis 7
Conclusion. Intensive care and PDs substitution(ATIIIc, ptl=27.000 IU, pt=30.000 IU
and differentamountsof fresh frozenplasma)improvedthe severecoagulationdefectsand stop the inhibitorconsumption.The parameterstypicallyelevatedin DIC-F-P-Sdecreased to normal values together with recovery. Both pts suffered transientlyfrom prltcoma or coma hepaticumrespectivelybut survived after long term intensivecare.
l
149 Thrombin generation, endothelial cell function and fibrinolytic potential in homozygous sickle cell disease and betathalassemia *GOUBRAN F, *NEVENE RAMSS1S, and **ATEF A *Clinical Pathology and **Pediatric Dep. Suez Canal University, Egypt Several studies have indicated that vaso-occlusion in sickle cell disease (SCD) is a multi-factorial process rather than a simple obstruction by sickled cells. Red blood cells from patients with h o m o z y g o u s betathalassemia behaves as procoagulant cells. W e tried to study thrombin generation and endothelial cell markers in relation to the fibrinolytic potential in SCD patients (n=9) during crisis and in the steady state as well as in h o m o z y g o u s beta-thalassemia patients (n= 11). The obtained results were compared to those o f age matched controls. Thrombin/antithrombin III complex (TAT), prothrombin fragment 1+2 (Fl+2) and thrombomodulin (TM) levels, as assessed by commercial E L I S A ' s , were mostly elevated in SCD during crisis followed by SCD in quiescent states and thalassemics when compared to controls. The activity of tissue type plasminogen activator (t-PA), plasminogen activator in-
hibitor type 1 (PAl-l), o~2-antiplasmin and histidine rich glycoprotein (HRG), determined by chromogenic substrate assays, were slightly increased during crisis in S C D and were not changed from control levels in the quiescent states and thalassemic patients. The amplitude o f thrombin generation was not asscociated with similar elevation in the fibrinolytic activity in SCD during crisis denoting that defective fibrinolysis participates in the vast-occlusive complication of SCD.