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

12 th Congress of the European Hematology Association
detection of the JAK2 V617F mutation were compared by testing blood
samples from 130 patients (115 CMPD, 15 non-CMPD) after informed
consent according to local clinical legacy had been taken. CMPDpatients
were classified as essential thrombocythemia (ET, n=56),
osteomyelofibrosis (MF, n=10), polycythemia vera (PV, n=47) and two
patients with unclassified CMPD. Methods. DNA was prepared out of
200 µL total blood and up to 200ng DNA were tested. On the one hand,
a qualitative PCR was used (Baxter EJ: Lancet 365, 2005) whereby mutated
as well as wildtyp JAK2 sequences are amplified in parallel. PCRproducts
were separated on agarose-gels. On the other hand, samples
were tested with a quantitative real-time PCR (Kroger N: Blood 109,
2007). In this method only mutated JAK2 molecules are detectable with
specific fluorescence-labelled TaqMan probes. The proportion of JAK2
V617F was calculated by a correlation with GAPDH . Results Corresponding
results with both assays were achieved in 34 negative patients
(15 ET, 4 MF, 1 PV, 14 non-CMPD) and 76 positive patients (25 ET, 5 MF,
45 PV, 1 unclassified CMPD). In patients with ET the median proportion
of mutated JAK2 was 15,7% (2,3%-52,7%), in MF 45,3% (28,8%-77%),
in PV 42% (4,4%-66,2%) and 17% in one patient with unclassified
CMPD. In addition, 10 patients (8 ET, 1 RARS-T, 1 unclassified CMPD)
with only weak mutated JAK2 DNA-bands on agarose-gels were clearly
positive in real-time PCR with a median proportion of mutated JAK2
of 4% (1,8%-8%). Finally, 10 qualitative PCR negative patients (8 ET, 1
MF, 1 PV) were positive in real-time PCR with a median proportion of
2,8% (1,1%-28,8%). Homozygosity was indicated in 10 Patients (8 PV,
1 ET, 1 MF) by JAK2 V617F proportions of equal or higher than 50%.
Overall, 41/56 (73%) patients with ET, 6/10 (60%) with MF and 46/47
(98%) with PV were JAK2 V617F positive. Conclusions. The quantitative
real-time PCR is more sensitive than qualitative PCR since only mutated
JAK2 DNA sequences are detectable. Ten of 44 (23%) qualitative
PCR negative patients were positive in real-time PCR. Differences in
the incidences of JAK2 V617F positive patients reported may, at least in
part, be caused by the detection-methods in use. In addition, the proportion
of mutated JAK2 differs in CMPD subgroups, indicating a possible
diagnostic value of the V617F quantification.
1545
THERAPY OF ANTIBODY-MEDIATED FACTOR VIII DEFICIENCY BY IMMUNOSUPPRESSIVE
THERAPY CONTAINING CYCLOPHOSPHAMIDE, DEXAMETHASONE AND RITUXIMAB:
A CASE REPORT
R. Thoedtmann, S. Rosenlechner, J. Unger, W. Patsch, R. Greil
University Hospital, SALZBURG, Austria
Background. Aquired factor VIII deficiency is a rare coagulation disorder
with an incidence of 1.34 cases per million population per year with
malignancy, post-partal status and autoimmune disease as most commonly
associated diseases. Besides control of active bleeding by substitution
with coagulation factor concentrates, immunosuppressive therapy
is the backbone of therapy. We report succesfull therapy of aquired
factor VIII deficiency with cyclophosphamide, dexamethasone and rituximab
in a patient with aquired factor VIII deficiency. Case. A 64 year
old male patient presented at our hospital with spontaneous hematoma
of the gluteal region and of both feet. Initial laboratory analysis revealed
prolonged partial thromboplastin time (ptt) (65 seconds), enhanced fibrinogen
activity (539 mg/dL) with normal prothrombin time and ATIII
level. Detailed analysis of coagulation parameters showed decreased factor
VIII activity (8.1%), increased von Willebrand factor antigen (> 250%)
and factor VIII inhibitor (35 Bethesda Unit, BU). Autoimmune diseases,
underlying malignant conditions, pulmonary diseases as well as inducing
concomitant medications were ruled out clinically. Thus the diagnosis
of aquired idiopathic antibody-mediated factor VIII deficiency was
established. The patient was treated with intravenous cyclophosphamide
750 mg/m2 day 1, oral dexamethasone 20 mg day 1 to day 7 and weekly
intravenous rituximab 375 mg/m2 day for 4 consecutive weeks. The
initial hematoma in the gluteal region as well as in both feet started disappearing
2 days after initiating of therapy. Normalization of factor VIII
activity was observed on day 49 after start of therapy. Concentration of
factor VIII inhibitor parallely decreased over time. New hematoma or
other signs of active bleeding did not occur. Significant toxicities observed
were an abscess in the gluteal region, which could be managed by surgical
means as well as systemic antibiotics; additionally, the patient developed
dyspnea with an interstitial pulmonary infiltration as underlying
condition. Further diagnostic work up revealed non-infectious pneumonitis,
that was judged to be rituximab-related. Administration of steroids
lead to improvement of the symptoms as well as CT-scan and functional
analysis based findings. One year after diagnosis the patient remained
546 | haematologica/the hematology journal | 2007; 92(s1)
stable. Especially new symptoms suggesting relapse of disease have not
occurred. Conclusion. We report a case of aquired factor VIII deficiency
that was successfully treated with an immunosuppressive regimen composed
of cyclophospamide, dexamethason and rituximab. The symptoms
of coagulopathy as well as the initially pathological laboratory findings
were released relativily fast, however toxicity of the regimen lead
to 2 consecutive hospitalisation of the patient, but could be resolved.
1546
THE IMPACT OF HIGH-DOSE SODIUM SELENITE THERAPY ON BCL-2 EXPRESSION
IN ADULT NON-HODGKIN`S LYMPHOMA PATIENTS :CORRELATION WITH RESPONSE
AND SURVIVAL
I. Asfour
Ain Shams University, CAIRO, Egypt
The present study was undertaken to explore the effect of administration
of high doses of sodium selenite on the expression of Bcl2 in patients
with non-Hodgkin’s Lymphoma. Fifty patients with newly diagnosed
non-hodgkin’s lymphoma were randomly divided into two groups,
Group A-1 received standard chemotherapy while group A-2 received
adjuvant sodium selenite 0.2 mg/kg/d orally for thirty days in addition
to chemotherapy after giving informed consent. Enzyme linked
immunosorbent assay(ELISA)was used ta assess BCL2 at the time of
diagnosis and after therapy in the two groups. Sodium selenite administration
resulted in significant decline of BCL2 level after therapy in group
A-2(8.6+ or - 6.9 ng/mL vs 6.9+ or - 7.9 ng/mL, p0.05), respectively. Down-regulation is not statistically significant
and therefore, the expression of each gene ranges within the
same values, either we examine AML patients with long-term complete
remission, or control individuals. Conclusion. Since our findings suggest
correlation between angiogenesis and apoptosis, additional work is now
desired to understand the role of VEGF and Survivin in the pathophysiology
of haematologic malignancies and the progression of AML. Further
studies are needed in order to determine whether VEGF and Survivin
could be used as prognostic markers, minimal residual disease
(MRD) indicators or promising cancer therapeutic targets.