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

treatment of acute promyelocityc leukemia and some other hematological
malignancies. Aim. To estimate the elimination speed of MLL
rearrangements in patients enrolled in phase I/II of single institutional
study. Methods. Since September 2003 4 patients from our clinic with
MLL rearrangements have been enrolled in this study. All of them have
had pro-pro-B (BI) immunophenotype. Leucocytes from bone marrow
were obtained. Nested PCR for MLL/AF4 (A. Borkhardt et al., 1994) and
MLL/ENL fusion genes (FG) were performed. Design of primers, probe
and real-time quantitative PCR (qRT-PCR) conditions for MLL/AF4 FG
detection was previously described (J. Gabert et al., 2003). β2-microglobulin
was used as control gene. Normalized copy number (NCN) of
MLL/AF4 was calculated. This value was multiplied by 10 000 000 and
Log10 was taken. In accordance with treatment design patient with
MLL/ENL has been treated by intermediate risk arm, while 3 others
infants with MLL/AF4 have been enrolled to the high risk (HR) schedule.
Treatment and diagnostics has been approved by Institutional Ethics
committee. Parents’ informed consent were obtained in all cases. Results.
Qualitative nested PCR revealed that 3 patients had MLL/AF4 and 1 had
MLL/ENL FG transcripts. In 2 patients with MLL-AF4 and 1 patient with
MLL/ENL total elimination of FG were detected after first course of
ATRA administration on day 43. FG were not detected by means of
nested PCR with sensitivity of 10-5. In 1 patient elimination of MLL/AF4
was found on the first day of protocol II after 7-th ATRA pulse. In this
patient qRT-PCR was performed on days 1, 15, 36 as well as first days
of HR blocks 1(1), 3(3), first day of protocol II. There was not available
material for qRT-PCR on first days of HR blocks 2(2), 1(4), 2(5), 3(6).
Although in all above mentioned time points nested PCR of MLL/AF4
FG transcripts was performed. On the day 1 Log10 NCN of MLL/AF4
was 4.77. Within induction therapy Log10 NCN values were almost
equal-2.36 on day 15 and 2.33 day 36, respectively. Significant reduction
was found after first course of ATRA, NCN was 1.95. After that NCN
was going down by degrees and on the first day of HR blocks 3(3) it was
0.39. Since the first day of protocol II MLL/AF4 FG transcript has been
detected neither in qRT-PCR nor in nested PCR. All subsequent PCR
examinations have not been detected the MLL/AF4 FG. Conclusions. It has
been proved the effectiveness of ATRA-content regimen for infants with
MLL/AF4 and MLL/ENL FG transcripts. Administration of ATRA led to
significant reduction of MLL/AF4 and MLL/ENL transcripts down to
undetectable level.
1246
DIFFERENT TYPES OF BONE MARROW INVOLVEMENT BY DIFFUSE LARGE B-CELL
LYMPHOMA: SPECIAL FEATURES AND RELATIVE BM CHANGES
E. Chigrinova, A. Pavlovskaya, M. Frenkel, I. Poddubnaya, N. Tupitsyn
N.N. Blokhin Cancer Research Center, MOSCOW, Russian Federation
Background. The aim of the study was to evaluate the special features of
bone marrow involvement by Diffuse Large B-cell Lymphoma (DLB-
CL), relative BM changes and possible immunomorphological discordance
with other involved sites. Methods. We studied bone marrow (BM)
samples (24 aspirates and 23 trephine biopsies) from 24 patients consecutively
diagnosed with DLBCL in a single institution during a 3-year
period (2003-2006). The main including criteria was prominent lymphoid
infiltration in BM trephine biopsy’s samples and atypical lymphoid
(blasts) cells presence or lymphocytosis > 28% in BM aspirate’s
slides. For the diagnosis of DLBCL the current WHO criteria was used.
14 (58%) cases were nodal and 10 (42%)-extranodal. Sites of the extranodal
disease were: soft tissues-4 cases, Waldeyer’s ring (WR)-2, GI tract-
2, primary BM/bone involvement-1, and breast involvement '1 case.
When available, clinical data at diagnosis, including components of the
International Prognostic Index (IPI), were reviewed. Selected group consisted
of 16-previously untreated patients (7 cases-low risk, 3-intermediate
high and 6 'high IPI groups), 3- currently treated, 2-relapsed and 3with
not knower clinical stage. Immunophenotyping BM study was performed
in 8 cases of aspirate (flow cytometry-FC) and in 17 of trephine
biopsy simples (immunohistochemistry). Panel of monoclonal antibodies
routinely included lineage-associated and immune markers, proliferation
and apoptosis markers, and more recently stage-specific markers
of B- and T-cell differentiation. Results. We observed 3 main types of BM
changes in selected group, according to complete trephine biopsy investigation:
diffuse blasts cells infiltration-10 cases (42%), T-cell rich like pattern
of involvement-5 cases (21%) and clear reactive T-cell patchy infiltration-9
(37%). Diffuse blast cell infiltration was associated with nodal
presentation (9/10), low T-cells count (immunohistochemistry detection
on BM trephine biopsies) high IPI or relapsed group patients. T-cell rich
like BM infiltration pattern in contrast to primary sites composed of
non-neoplastic T cells and only 10 ng/mL (p=0,017; Odds ratio=10,57) and serum lactate
>3 mmol/L (p=0,04; Odds ratio=16) are correlated with occurrence
of septic shock. Serum level of PCT using semi-quantitative test have
equivalent diagnostic and prognostic value for the diagnosis of infection
and for outcome as CRP and serum lactate in neutropenic patient.
1248
ALLOGENEIC HAEMOPOIETIC STEM CELL TRANSPLANTATION IN PATIENTS AT HIGH RISK
FOR TOXICITY USING TREOSULFAN/CYCLOPHOSPHAMIDE±ALEMTUZUMAB
CONDITIONING
A. Fyfe, A.N. Parker, I.G. McQuaker, I.M. Franklin, T.L. Holyoake,
A.D. Clark
Glasgow Royal Infirmary, GLASGOW, United Kingdom
Background. Allogeneic transplantation is associated with well documented
toxicity. Treosulfan is an alkylating agent which has been reported
as offering reduced toxicity compared with busulfan without compromising
engraftment or increasing relapse rates. In patients thought to
be at high risk from toxicity associated with standard radiotherapy based
myeloablative conditioning, but requiring intensive conditioning, we
used Treosulfan (14 g/m2 D -6 to -4), Cyclophosphamide (60 mg/kg D -
3 to -2) and for those with volunteer unrelated or mismatched sibling
donors, MabCampath (10 mg bd iv D -6 to -1). Methods. We retrospectively
evaluated 12 patients (median age 42 range 25 to 50) who received
this regimen at our institution between 2001 and 2006 with a median follow
up of 398 days (range 64-1922). The patients were all at high risk
for relapse and had the following diagnoses AML CR1(3), AML CR2(2),
haematologica/the hematology journal | 2007; 92(s1) | 453