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

12 th Congress of the European Hematology Association
smoldering myeloma that reaches complete remission after HAART.
According to this clinical observation, if plasma cell dyscrasia is diagnosed
in an HIV-patient, physicians should be aware of the possible
benefit of HAART, and reevaluate the disease after immune restoration
and viral load control.
1349
TREATMENT OF CYTOMEGALOVIRUS REACTIVATION DURING THERAPY
WITH ALEMTUZUMAB IN PATIENTS WITH CHRONIC LYMPHOCYTIC LEUKEMIA
A DILEMMA TO SOLVE
M. Wach, A.D. Dmoszynska, E.W-S. Wasik-Szczepanek, M.C. Cioch
University School, LUBLIN, Poland
Alemtuzumab (Campath), monoclonal antibody, anti-CD52, is a new
drug for a treatment of chronic lymphocytic leukemia (CLL), that has an
importance as monotherapy and also in combination with chemotherapy.
Among side effects of alemtuzumab reactivation of
cytomegalovirus (CMV) and its therapeutic management seems to be an
important problem. Therapeutic standard in reactivation of CMV is to
stop the treatment with alemtuzumab and to start the treatment with
ganciclovir until CMV negativity. Here it is presented own experience
concerning CMV reactivation in ten patients with progressive CLL, treated
with alemtuzumab as a first-line treatment. They were three women
and seven men, aged 53-75 years (median age -59.7years). Two of them
were in stage I according Rai classiffication, five in stage II, two in stage
III and one in stage IV. Median time from diagnosis to treatment varied
between 1 month to 101 months (median time-19.7months).Absolute
lymphocyte count before treatement was 56.7G/l to 228.0G/l (median
count-107.0G/).All patients were CMV seropositive before treatment
with alemtuzumab and anti-CMV IgG titre was 1:230 to 1:49 000 (median
'1:16 653).The patients were reported to have negative anti-CMV
IgM and also serum CMV-DNA assay by PCR method was below 500
copies/mL. Patients received alemtuzumab at a dose 30mg I.V. three
times weekly during 12 weeks. Prophylaxis of infection consisted with
aciclovir at a dose 800 mg daily and co-trimoxazole at a dose 1920 mg
three times weekly during treatment period and two months after stopping
the therapy. Serum quantitive CMV-DNA was estimated every
week using PCR method and patients were underwent physical examination
3 times weekly. During treatment period in eight patients (80%)
reactivation of CMV occurred, in one case symptoms of CMV disease
was observed (fever is the only symptom) and in one patient neither
reactivation nor CMV disease occurred. The CMV reactivation appeared
between 3-8 weeks of treatment . Serum CMV-DNA titre was; 700 to
19 000 copies/mL: only in one patient it was 380 000 copies/mL (this
patient had symptomatic CMV disease and received the treatment with
ganciclovir). We took into consideration the treatment with ganciclovir
in all patients but the decision depended on the presence of CMV symptoms,
titre of serum CMV-DNA during reactivation and dynamics of
increasing CMV-DNA titres which were estimated every week. Two
patients received the treatment with ganciclovir; one of them had CMV
disease, seven patients despite positivity of CMV-DNA had not received
any treatment. None of these seven patients had symptoms of CMV disease.
In all these patients during 1 to 4 weeks spontaneous decreasing
of CMV-DNA were observed. In five patients with a low titre of serum
CMV-DNA, without increasing this titre, the treatment with alemtuzumab
was not interrupted. It is concluded, that in the case of CMV
reactivation in CLL patients treated with alemtuzumab, without symptomatic
CMV disease, very careful clinical observation , every-week
CMV-DNA assay and prophylaxis with aciclovir is a sufficient management.
In other patients with CMV reactivation, particulary exposed on
CMV disease (even slight symptoms of CMV disease, high rate of CMV-
DNA titre observed during reactivation and high rate of anti-CMV IgG
before the treatment) the therapy with ganciclovir is recommended.
486 | haematologica/the hematology journal | 2007; 92(s1)
1350
MOLECULAR SCREENING OF PDGFRA AND PDGFRB GENES IN KIT AND FLT3 NEGATIVE
CORE BINDING FACTOR LEUKEMIAS
R. Cairoli, 1 A. Trojani, 1 C.B. Ripamonti, 1 ,S. Penco, 1 A. Beghini, 2
G.P. Nadali, 3 F. Elice, 4 A. Viola, 5 C. Castagnola, 6 P. Colapietro, 2
G. Grillo, 1 P. Pezzetti, 1 E. Ravelli, 7 A. Marocchi, 1 M.C. Patrosso, 1
A. Cuneo, 8 F. Ferrara, 5 F. Rodeghiero, 4 M. Lazzarino, 6 G. Pizzolo, 3
E. Morra1 1 Ospedale Niguarda, MILAN; 2 University of Milan, MILAN; 3 University of
Verona, VERONA; 4 San Bortolo Hospital, VICENZA; 5 Cardarelli Hospital,
NAPLES; 6 University of Pavia, PAVIA; 7 Niguarda Hospital, MILAN; 8 S.Anna
Hospital, FERRARA, Italy
Mutations involving KIT and FLT3 genes, encoding tyrosine kinase
(TK) membrane receptors, are frequently detected in core binding factor
leukemia (CBFL) adult patients and are reported as mutually exclusive.
PDFGRA and PDGFRB encode class III TK receptors and are
involved both in physiological processes, such as fibrosis, and in the
pathogenesis of haematological and solid tumours. Mutations in
PDGFRA are found in gastrointestinal stromal tumours (GIST), rarely in
synovial sarcomas (SSs) and in malignant peripheral nerve sheath
tumours (MPNST), whereas FIP1L1-PDGFRA fusion product occurs in
systemic mastocytosis associated with eosinophilia, in idiopathic hypereosinophilic
syndrome, chronic eosinophilic leukemia and in polycythemia
vera patients. Many different PDGFRB chimeras are described
in BCR-ABL negative chronic myeloproliferative disorders. In order to
detect PDGFR mutations in CBFL, we screened 35 KIT and FLT3 negative
patients by specific SSCP analysis and direct sequencing. For analysis
of the juxtamembrane and TK domains of PDGFRA, exons 9, 11-15,
17-20 were amplified. We studied exons 12 and 18 for analysis of the
TK domain of PDGFRB. PCR were loaded on non denaturing gradient
gels (5-20%) at two different temperature conditions (12°C and 23°C)
for 20 hours. Samples displaying abnormal migration patterns underwent
direct sequencing with forward and reverse primers. Three types
of single-nucleotide variations (SNP) were detected in PDGFRA gene: in
exon 13 (rs10028020-SNP ID) 5 out of 35 patients showed a GCG>GCA
change in heterozygous form; in exon 18 (rs2228230-SNP ID) 5/35
patients displayed a GTC>GTT change in heterozygous form and in
exon 12 (rs1873778-SNP ID) 4/35 patients showed a CCA>CCG change
in homozygous form. Moreover, exon 13 and exon 18 polymorphisms
were both present in 3 patients. All three SNPs in PDGFRA gene were
previously described. No molecular alteration was detected in PDGFRB
gene. In conclusion, no pathogenic mutation in PDGFR genes was
detected among our KIT and FLT3 negative CBFL patients and further
molecular investigations will be needed to better clarify their genetic
characteristics.
1351
ASSOCIATION OF CD38 EXPRESSION AND DIAGNOSTIC IMMUNOPHENOTYPIC SCORE
IN B-CELL CHRONIC LYMPHOCYTIC LEUKEMIA
C. Kartsios, C. Kartsios, E. Yiannaki, M. Ditsa, E. Katodritou,
P. Konstandinidou, D. Mihou, E. Sotiriadou, K. Zervas, D. Markala
Theagenion Cancer Hospital, THESSALONIKI, Greece
Background-aims. The typical B-cell chronic lymphocytic leukaemia
(B-CLL) phenotype is CD5 + , CD23 + , FMC7- , dim expression of surface
Ig (sIg) and weak/absent expression of membrane CD22 and CD79b.
Because there is no specific marker for B-CLL, a composite phenotype
considering the above markers compounded into a scoring system, helps
to distinguish CLL from the other B-cell malignancies. According to this
system, 92% of B-CLL cases score 4 or 5, 6% score 3 and 2% score 1 or
2. CD38 is a membrane protein that marks cellular activation and it is
now considered as an independent prognostic marker of outcome. We
study the possible relationships between a prognostic marker (CD38)
and the immunophenotypic score-consisting antigens. Patients and Methods.
From 2000 to 2006, 176 previously untreated B-CLL patients [M/F:
99/77, median age: 68 (36-87) years] were diagnosed at our centre. The
lymphocyte morphology was typical in 171 cases (97%) and the majority
of the patients [160/176, (91%)] were classified as Rai stage 0-2.
Three-color flow cytometric analysis was performed on their blood samples
and the reactivity with CD2, CD5, CD19, CD20, CD22, CD23,
CD25, CD38, CD79b, FMC7, and immunoglobulin kappa and lambda
light chains, was analyzed. Positivity for antigen expression was set at
30% while two cut-off points (7% and 30%) were used for CD38
expression. Mann Whitney U test and Spearman’s rho test were used for