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

1691GA and 1 homozygous 1691AA. The prevalence of FV 1691 mutation
was found to be 2.6% with an allele frequency of 1.49%. Factor V
leiden was found in 3 males and 3 females as heterozygous and in a
male as homozygous. Summary/Conclusions. For the first time the allele
frequencies for prothrombin G20210A and FV Leiden for one of ethnic
groups (Kurds) living in Iran is reported. The frequency was found for
FV 1691A allele is in the range of those reported for Caucasoid subpopulations
(1-8.5%) and is similar to some of Asian countries including
North India and Saudi Arabia. However, the frequency of prothrombin
20210A allele in our population is higher than those reported for some
Middle East countries. Our study indicates that factor V Leiden, and prothrombin
gene 20210A are not rare in Iranian population and need to be
considered in patients with venous thrombosis.
1225
CHANGES OF MDM2 AND P53 PROTEINS EXPRESSION IN RELATION TO EARLY
TREATMENT RESPONSE IN CHILDREN WITH ACUTE LYMPHOBLASTIC LEUKEMIA
T. Ociepa, E. Maloney, E. Kamienska, T. Urasinski, E. Urasinska
Pomeranian Medical University, SZCZECIN, Poland
Background. Several pieces of evidence suggest that ineffective apoptosis
can be one of the main causes of treatment failure in childhood
acute lymphoblastic leukemia (ALL). P53 is the pivotal gene in controlling
apoptosis of human cells. MDM2, the endogenous inhibitor of apoptosis
is an oncoprotein which downregulates the functional activity of
p53 protein. Thus, increased activation of MDM2 can be among factors
responsible for treatment failure in childhood ALL. This study aimed to
evaluate changes in MDM2 and p53 expression in peripheral blood
mononuclear cells collected prior to and after 6 and 12 hours of prednisone
administration, and the relation between them and early treatment
response. Patients. The study was comprised of 35 children (aged
6-192 months, median 95 months) with newly diagnosed ALL. The diagnosis
was based on morphologic examination of bone marrow smears
stained with MGG and by results of flow cytometric immunophenotyping
of bone marrow leukemic cells. All children were treated according
to the ALL BFM 90 protocol. Patients with absolute leukemic cell counts
of less than 1000/mL of peripheral blood after 7 days of prednisone treatment
(plus one intrathecal injection of Mtx) and with leukemic cells
below 5% in bone marrow smears aspirated on day 15 were classified
as early good responders (n=24), whereas the remaining patients were
classified as early poor responders (n=11). The follow up time was 1-42
months (median 21 months). Methods. Peripheral blood mononuclear
cells were collected prior to and after 6 and 12 hours from prednisone
administration. Cytospin preparations of these cells were stained with
mouse monoclonal anti-MDM2 antibodies (DakoCytomation) followed
by goat anti-mouse antibodies conjugated with APC (Molecular Probes)
and mouse monoclonal anti-p53 antibodies conjugated with FITC
(DakoCytomation) respectively. Nuclear DNA was stained with propidium
iodide (PI). MDM2 - associated long red fluorescence and p53associated
green fluorescence and were measured by laser scanning
cytometer (LSC, CompuCyte, USA). In order to assess the rates of apoptotic
cells respective slides were stained with polyclonal rabbit anti-PARP
p85 fragment followed by FITC conjugated swine anti-rabbit antibody.
Cell expressing p89 fragment of PARP were considered apoptotic. Red
fluorescence of DNA-bound PI was used as a contouring parameter. Values
of long red integrated fluorescence and green integrated fluorescence
were recorded as .FCS 3.0 files by WinCyte 3.4 software. Results. The
mean pretreatment values of p53 expression in both groups did not differ,
whereas mean pretreatment values of MDM2 expression were significantly
higher in the group of early poor responders (p=0,03).After 12
hours from prednisone administration mean values of p53 expression
were significantly higher in the group of good early responders (p=0,01).
Higher expression of p53 in these patients was associated with significantly
higher rates of apoptotic cells as compared with poor early treatment
responders (p=0,03). Moreover, p-EFS in the group of early treatment
responders was significantly higher than for the remaining patients
(1,0 vs 0,635; p=0,002). Conclusions. These data seem to indicate that pretreatment
overexpression of MDM2 protein may contribute to poor early
treatment response, thus influencing the outcome for these patients.
12 th Congress of the European Hematology Association
1226
EARLY AND LATE COMPLICATIONS OF CHEMOTHERAPY IN ACUTE LYMPHOBLASTIC
LEUKAEMIA. A SINGLE PAEDIATRIC INSTITUTION EXPERIENCE
A. Mangione, 1 G. Menna, 1 N. Marra, 1 A. Barberi, 1 A. Antignano, 1
L. Foggia, 1 F. Cecere, 2 V. Poggi, 1 R. Parasole1 1 2 „Santobono-Pausilipon„ Hospital, NAPLES, Italy; Ruggi d’Aragona Hospital,
SALERNO, Italy
Background. Improvement in survival rates after acute lymphoblastic
leukaemia (ALL) have sharpened the need for research on adverse events
after treatment. These sequelae can have negative effects on quality of
life of survivors and when severe, they are cause of morbidity. The current
study assesses the incidence of early and late effects by a single paediatric
institution. Patients and Methods. Between December 2005 and
January 2007, one hundred and eighty-eight patients, with off-therapy
ALL, were followed-up by a specialized multidisciplinary team. The
median age of children at diagnosis was 4 years and 6 months (range 3
months-15 years and 6 months), 96 males and 92 female. The median
time of follow-up was 51 months (range 3 months-19 years and 6
months). One hundred and eighty-three were treated with ALL-AIEOP
protocols, three with infant-ALL protocol and two patients with
relapsed-ALL ones. According to immunophenotype, 155 (82,5%) were
B-ALL and 16 (8.5%) T-ALL, 1 biclonal (0.5%) and 16 (8.5%) were
unclassified. The cranial irradiation was performed on 33 (17.5%)
patients and only one child received testis radiotherapy. All patients
underwent a periodic clinical, haematological, endocrine, viral, cardiac
and radiological checks. A careful long-term follow-up was performed
for a rapid identification of organ-specific sequelae or secondary malignancy.
Results. Seventy-two (38.3%) patients presented at least one early
complication and 20 (27.7%) two or more effects. The earliest event
were infections (40.6%), followed by neurological sequelae (30.8%).
Long-term complications were observed in 121 (64,4%) patients, prevalently
endocrine sequelae (51.9%) (especially obesity) and dental complications
(14.5%). Few data were available on fertility or sexual dysfunctions,
due to young age of patients; only three pregnancies were
observed in two female patients. In a median follow-up of 51 months
after treatment, only one secondary malignancy was observed, an
abdominal ganglioneuroma, completely surgically removed. During routine
controls, two relapses (testis and optic nerve) were precociously
diagnosed. Conclusions. These results suggest that new therapeutic
approaches in acute lymphoblastic leukaemia have significantly
improved survival rate but it is burdened by high incidence of late effects
that sometimes affect negatively the quality of life of survivors. Then,
we believe that the patient and family have to be well informed about
all possible complications related to treatment. Duty of clinicians is to
perform a careful, prolonged follow-up for a precocious identification
and treatment of long-term effects. Future studies should focus on strategies
aimed at reducing late sequelae, probably using treatment regimen
at reduced toxicity.
1227
HODGKIN LYMPHOMA IN MUTATED ZAP-70 NEGATIVE B-CELL CHRONIC LYMPHOCYTIC
LEUKEMIA: A CASE REPORT
P. Niscola, 1 M.I. Del Principe, 2 M. Palombi, 1 S. Fratoni, 3 D. Piccioni, 1
L. Maurillo, 1 P. De Fabritiis, 1 S Amadori, 2 G Del Poeta1 1 Tor Vergata University, S.Eugenio Hosp, ROME; 2 Tor Vergata University,
ROME; 3 Pathology Dept, Sant’Eugenio Hospital, ROME, Italy
Background. The transformation of B-cell Chronic Lymphocytic
Leukemia (CLL) into more aggressive lymphoproliferative disorders in the
form of Hodgkin lymphoma (HL) is a rare event portraying a very poor
prognosis. From the analysis of CLL biologic features predisposing to this
complication arise some concerns which can be elucidated by the following
presentation of a case recently observed by us. Case Report. A 60 year
old man was diagnosed as having a classic B-CLL (stage II according to
Rai classification) in November 1990 and followed for 11 years until the
progression to stage IV. The molecular analysis of genes encoding for the
variable region of immunoglobulin heavy chains (IgVH) revealed a mutated
status (VH: 4-61; D: 7-27; JH: 4;% of mutation: 5.4). CD38 and ZAP-
70 determined by flow cytometry were negative and FISH analysis
showed no abnormalities. He underwent standard treatment with fludarabine
achieving a complete response (CR) consolidated (January 2002)
by four weekly standard doses of rituximab. CR was maintained until
May 2006 when the patient’s clinical conditions suddenly worsened.
Indeed, he presented general malaise, emaciation, mild splenomegaly
without palpable lymph nodes and severe hypocromic anemia. A CT-
haematologica/the hematology journal | 2007; 92(s1) | 447