12th Congress of the European Hematology ... - Haematologica

12 th Congress of the European Hematology Association
ured before cryopreservation. Results. The mean total nucleated cells,
viable CD34 + cells, and CFU colonies recovery was 47%, 72% and 55%
respectively for the Voluven washing solution. The mean total nucleated
cells viable CD34 + cells, and CFU colonies recovery was 37%, 54%
and 60% respectively for the albumin, NaCl and ACD washing solution.
The comparison of the total nucleated cells viability and the CD34 viability
between the Voluven solution and the albumin, NaCl and ACD
solution showed respectively 55% and 45% viable TNC, and respectively
78.8% and 68% viable CD34 + cells. Conclusions . The washing of cord
blood with a solution containing Voluven improves significantly TNC
and CD34 + cells recovery and viability. No significant difference was
observed on CFU colonies recovery.
1190
NEW CHROMOSOME ABERRATIONS IN B-CELL CHRONIC LYMPHOCYTIC LEUKEMIA
R. Boesmueller, 1 S. Moshir, 2 U. Jaeger, 3 C. Fonatsch4 1 MUW, VIENNA, Austria; 2 MUW, Department of Medical Genetics, VIEN-
NA, Austria; 3 MUW, Department of Internal Medicine I, VIENNA, Austria;
4 MUW, Department of Medical Genetik, VIENNA, Austria
B-cell chronic lymphocytic leukemia (CLL) is a common leukemia
that more frequently affects elderly people. The clinical course of CLL
is variable and in early stages a prediction of disease progression is difficult.
Specific karyotype deviations are important prognostic factors, but
within the group of patients with a normal karyotype that is in general
associated with a good prognosis a number of patients show an aggressive
clinical course. The aim of our study is the detection and characterization
of new chromosome aberrations by classical and molecular cytogenetics.
In comparison to fluorescence in situ hybridization (FISH) conventional
cytogenetic analysis by classical chromosome banding plays
a minor prognostic role in CLL due to the low proliferation rate of malignant
B-cells in vitro. As traditionally used sets of mitogens result in poor
proliferation response of leukemic B-cells, new cultivation techniques
using optimal mitogen combinations (OMC: TNFα + IL-2, SAC + IL-2,
TNFα+ TPA10) and immunostimulatory CpG-Oligodinucleotide DSP 30
(COD) plus IL-2 were established in order to enhance the yield of
detectable chromosome aberrations in CLL cells. Successful metaphase
stimulation by culturing the cells with OMC and/or COD plus IL-2 was
observed in 45 CLL cases. Most of these karyotypes showed the same
aberrations as obtained by parallel performed interphase FISH. In addition
six novel chromosomal aberrations were identified and characterized.
Detailed depiction of novel aberrations was possible by FISH using
whole chromosome painting probes, centromere-specific, single copy
probes and/or multicolor FISH. The relevance of these novel chromosome
anomalies in CLL detected by using refined culture techniques
and the combination of classical chromosome banding and molecular
cytogenetic analyses with respect to other prognostic parameters and to
the clinical course will be examined.
1191
HEMOSTASIS EVALUATION IN PATIENT WITH ACUTE RENAL FAILURE (ARF) DURING
HEMODIALYSIS
F. Valeri, P. Schinco, A. Valpreda, C. Aguzzi, A. Borchiellini,
E. Beggiato, F. Bermond, M. Boccadoro
Molinette Hospital Turin, TORINO, Italy
Background. Critically ill patients often undergo ARF, for which
hemodialysis (HD) is indicated; in about 20% of cases coagulation of the
dialytic filter occurs despite anticoagulant therapy and the procedure
must be stopped. So far, no prognostic factor has been identified to recognize
patients at risk of dialyser clotting. Aims. We analysed the role of
three different anticoagulation strategies on the occurrence of filter clotting,
and we tried to identify a number of hemostatic parameters predictors
for dialyser coagulation. Methods. 22 critically ill patients were
studied before and during HD; 7 were anticoagulated with unfractionated
heparin (UFH), 4 dermatan-sulfate (DS), 11 received no anticoagulation
because at bleeding risk. Factor VIII:C (VIII:C), von Willebrand factor
(vWF), P-selectin, Protein C (P:C), antithrombin III and thromboelastography
(TEG) were assayed at T0 (before start of filtration), T1 (4
hours after start), T2 (7 hours after start or at filter clotting). We performed
qualitative analysis of vWF by agarose gel electrophoresis, aimed
to identify the presence of abnormal multimeric forms (HMWM), at all
times. Platelet function was also analysed with PFA-100 instrumentation,
in order to find out if this methodology could supply information on
platelet (hyper)reactivity. PFA-100 analysis was performed at all times.
Anti-heparin/PF4 antibodies (HIT-Ab) were assayed at T0 in all patients.
436 | haematologica/the hematology journal | 2007; 92(s1)
Results. 4 patients out of 22 underwent filter coagulation before end of
procedure; of these, 3 were treated with UFH, 0 with DS and 1 was not
anticoagulated; 2 of these were the only ones positive for HIT Ab. VIII:C
level was significantly increased in all patients at baseline and remained
unvaried during HD. Mean VIII:C level was 3.03 U/mL± 1.30 (vs 1.00±
0.40 normal range). vWF was also increased (mean 5.72 U/mL± 2.75 vs
1.00 U/mL± 0.40 normal range) and interestingly in 11 out of 22 patients
qualitative analysis revealed circulating HMWM. P-selectin was raised
in all patients and did not vary throughout HD ( 3920 ng/mL± 34 at T0,
1702 ng/mL±1926 at T1, 2108 ng/mL±1012 at T2 vs 137±18 ng/mL n.v.
). In most patients (14/22, 64%) P:C was heavily reduced, often below
30% ( 0.28 U/mL±0.9 ). The PFA-100 instrumentation showed to be
unable to detect cell hyperreactivity, and closure times did not correlate
with filter coagulation. Conclusions. Dialyser clotting in critical patients
undergoing HD seems to be a polyfactorial event: indeed pre-term filter
coagulation occurred in patients with multiple hemostasis pro-thrombotic
derangements and was more frequent among those with the presence
of HMW multimers in plasma. Assay of plasma coagulation
inhibitors and promotors may be a useful tool to asses the hemostatic
balance of such patients and to identify those at higher risk of filter preterm
coagulation. A scoring system is proposed for this purpose.
1192
INTRATHECAL RITUXIMAB IN COMPLEX TREATMENT OF REFRACTORY CD20-POSITIVE
PRIMARY CNS LYMPHOMA
I. Savic, I. Urosevic, S. Popovic
Clinical Centre Novi Sad, NOVI SAD, Serbia
Background. Rituximab binds specifically to the antigen CD20 which
is expressed on more than 90% of B-cell NHL and primary CNS lymphoma
(PCNSL) but is not expressed by normal neurons and glia in the
brain. Patients and methods. Three patients with recurrent CD20+ PCNSL,
IELSG score 1-2, were treated. High-dose methotrexate (MTX) was the
basic systemic chemotherapy in one patient while others received 6
cycles of R-CHOP plus intrathecal therapy (MTX, AraC and hydrocortisone)
and 40 Gy whole brain irradiation. All patients showed out MRI
evidence of residual brain parenchymal disease. Nine planned intrathecal
injections of rituximab in a dose of 20 mg each were given over 5week
period. Rituximab was administered in 2 mL of normal saline during
2 minutes. Tumor response was assessed by weekly CSF cytology
and immunocytochemistry, neurological examination twice weekly, and
MRI scanning at 5th week compared with baseline. MRI scan, physical
and neurological examination were repeated at 9th week (4 week after
final injection). Results. Intrathecal administration of rituximab was well
tolerated and all patients exhibited cytological and biochemical response
without CD20 + B cells detectable in CSF and no MRI evidence of PCNSL.
Consolidation therapy included 12,5 mg MTX and 100 mg hydrocortisone
intrathecaly weekly for 6 weeks. All patients are still in complete
remission without CD + B cells in CSF, and normal brain MRI. One patient
with IELSG prognostic score 1 is still receiving intensive MTX and
Cytarabine treatment which will be followed by high-dose chemotherapy
(BEAM) and ASCT. Conclusions. Overall survival of patients ranged
from 15 to 30 months (15, 18, and 30 months, respectively) and there is
no progression of disease after intrathecal rituximab therapy. These suggest
that intrathecal application of rituximab may have a role in the
treatment of refractory PCNSL.
1193
A MODIFIED BEAC-CONDITIONING PROTOCOL: A SAFE AND EFFECTIVE THERAPY
FOR HIGH RISK ALL
O. Krieger, H. Kasparu, M. Binder, J. König, S. Machherndl-Spandl,
O. Zach, D. Lutz
Elisabethinen Hospital, LINZ, Austria
Stemcelltransplantation is an established strategy for high-risk ALL.
Conventional conditioning schedules (eg.TBI+cyclophosphamide and
others) do have a substantial toxicity which has to to be regarded seriously.
Therefore we used a 7 days lasting chemotherapy conditioning
regimen consisting of BCNU (300 mg/m2 ), Etoposide (800 mg/m2 ), ARA-
C (3 g/m2 ), Cyclophosphamide (120 mg/kg) and 2-Chlorodeoxyadenosine
(24 mg/m2 ) for these patients. For unrelated donor-transplantations
(UD-Tx) ATG was added for two days. Patients. So far, 12 adult patients
(6 male, 6 female) with a Ph + pos. ALL (8), Pre- T-ALL (2), Pro-B ALL (1)
in first CR and 1 relapsed cALL, with a median age of 41 years (19-53)
were treated. In 6 patients a HLA ID Allo-PBPCT was performed, 1 HLA