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

nomenon is unclear. It has been suggested that the observed T cell clonality
may simply be related to the normal aging process or that it may
result from the immune response to malignant cells or viral infection.
During normal T cell maturation in the thymus, the T cell receptor (TCR)
genes undergo a complex process of V(D)J segment rearrangement to
produce a wide repertoire of antigen specificity. As it is an early event
in T cell development, TCRG gene rearrangement is commonly used as
a marker of clonality. The TCRB gene is also routinely investigated due
to its greater combinatorial diversity. Lack of sufficiently sensitive detection
methodology is often a limiting factor in the discrimination between
clonal and polyclonal T-cell populations. Aims. The aim of this study
was to use high resolution heteroduplex gel and fluorescence GeneScan
analysis to screen for TCRG and TCRB gene rearrangements respectively
in a cohort of B-CLL patients and to ascertain if the clonal rearrangement
occurred in either B cells or T cells. Methods. B and T cells were purified
from peripheral blood using a CD19 + or CD3 + magnetic bead system
(autoMACS). DNA was extracted from purified B or T cells from 34
B-CLL patients and the TCRG and TCRB genes were amplified by BIO-
MED-2 multiplex PCR assays (InVivoScribe Technologies). Heteroduplex
and GeneScan analysis were then performed to detect monoclonal
T-cell expansion. Results. Clonal TCR gene rearrangements were detected
in11/34 (32%) of cases in purified T cell fractions. No clonal TCR gene
rearrangements were found in purified B cell fractions. Clonal TCRB
rearrangements were found in 10/34 (29%) and clonal TCRG in 8/34
(23%). In most instances a weak clonal pattern was observed. Clonal
TCR rearrangements were detected in 7 males and 4 females. Five
patients had mutated IGVH genes, while the remaining 6 possessed
unmutated IGVH genes. Seven patients presented with more advanced
clinical stage (Binet B or C), and 8 patients received treatment.
Summary/conclusions. These results are in agreement with previous work
demonstrating that T-cell clonal/oligoclonal expansions occur frequently
in B-CLL patients. However this is the first report to demonstrate that
the clonal expansions occur in T cells and not in B cells. In most instances
a weak clonal pattern was observed which was probably due to minor
clonal T-cell populations. It is possible that T-cell clonality may be associated
with a poorer prognosis in this disease category. Of interest, we
found that 8/11(73%) patients in our B-CLL sub-group with T-cell expansion
also had advanced stage disease and/or unfavourable molecular
(IgVH gene usage/mutational status) markers. We are currently investigating
the possibility of a specific antigenic stimulus resulting in this
clonal T-cell expansion.
1308
INCIDENCE OF SECOND NEOPLASMS IN CHRONIC LYMPHOCYTIC LEUKAEMIA:
INFLUENCE OF PROGNOSTIC BIOMARKERS
M.A. Catherwood1 , R.J. Middleton2 , T.C.M. Morris1 , H.D. Alexander1 1 Belfast City Hospital Trust, BELFAST, Northern Ireland; 2 Queen’s University
of Belfast, BELFAST, United Kingdom
Background. B-cell chronic lymphocytic leukaemia (B-CLL) is the most
common leukaemia in the west, with a median survival of approximately
10 years. It is a heterogenous disorder with a highly variable clinical
course and it is well recognized that patients with CLL are at a higher
risk of developing second malignancies when compared to age and sexmatched
controls. IGVH mutational status provides prognostic information
in CLL, with unmutated IGVH status conveying a poor prognosis
in comparison to mutated IGVH genes. Aims. To determine what influence
the mutational status of the IGVH gene, gene usage and acquired
cytogenetic aberrations play in the development of a second malignancy
in patients with CLL. Methods. Three hundred and twenty patients
were recruited from the Haematology Outpatient Clinic and surrounding
regional hospitals. Clinical staging, immunophenotyping, lymphocyte
doubling time (LDT) and time to treatment (TTT) were available on
all patients. IGVH mutational status and gene usage were determined
using multiplex BIOMED-2 primers (InVivoScribe Technologies) and
sequence analysis. Chromosomal abnormalities were determined using
interphase fluorescence in situ hybridisation (FISH). Results. Results
revealed that solid tumours occurred in 57 patients (18%) of which 24
were second malignancies and 33 were malignancies occurring before
the diagnosis of CLL. The second malignancies occurred in the following
sites: skin (14) lung (4), breast (2), bowel (2), prostate (1), and 1 case
of stomach cancer. Median time from diagnosis of CLL to second malignancy
was 48 months (10-204 months). Eighteen patients were male
and six were female. Interestingly 14 patients had mutated IGVH genes,
while the remaining 10 possessed unmutated IGVH genes. No specific
gene usage was associated with second malignancies. Material for FISH
analysis was available on all cases. The mutated case showed no
12 th Congress of the European Hematology Association
detectable abnormality (n=6) and biallelic/del13q14 (n=8). The unmutated
sub-group consisted of: biallelic/del 13q14 (n=6) and trisomy 12 (n=4).
Nine patients presented with more advanced clinical stage (Binet B or C),
and 12 patients received treatment. Summary/conclusions. It is well documented
that patients with CLL have an increased risk of developing second
malignancies, over and above that expected for a matched age
group. This may be due to several factors, either singly or synergistically,
including defective immune surveillance due to an impaired adaptive
immune system associated with CLL, or it may be treatment related,
associated with, for example, the use of alkylating agents such as chlorambucil.
Our data shows that the second malignancies occur in both
treated and untreated patients. Secondary neoplasms are also found in
mutated and unmutated cases. Our data suggests that the development
of a secondary neoplasm is dependent on the length of disease duration
rather than therapy duration.
1309
THE REVERSE CORRELATION OF CD38 + /CD62L- VERSUS CD62L + /CD38- IN B-CELL
CHRONIC LYMPHOCYTIC LEUKEMIA
G. Baxevanos, 1 L. Dova, 2 H. Kapsali, 3 M. Ovrenovits, 4 H. Dokou, 2
N.I. Kolaitis, 4 K.L. Bourantas, 3 G. Vartholomatos2 1 University Hospital of Ioannina, IOANNINA; 2 University Hospital/Molecular
Biology, IOANNINA; 3 University Hospital/Haematology Dpt, IOANNINA;
4 University Hospital/Haematology Lab, IOANNINA, Greece
Background. CD38 is a transmembrane glycoprotein expressed on the
surface of leukemic cells in a significant percentage of patients with Bcell
chronic lymphocytic leukemia (B-CLL). A recent study suggested
that CD38 expression has negative prognostic value in CLL.The leukocyte
selectin (CD62-L) is expressed on hematopoietic stem-progenitors
and mediates adhesive interactions with other receptors. Aims. The aim
of this work was to study the correlation between the expression of
CD38 + and CD62L + on the pathogenic B-CLL cells. Methods. Peripheral
blood samples from 40 patients with B-CLL were analyzed by flow
cytometry for CD38 and CD62L expression on CD5 + CD19 + leukemic
cells. We chose to consider only typical B-CLL patients, having a score
of 4-5, according to the classification of typical/atypical B-CLL proposed
by Matutes et al.1994. The analysis was performed by the FACScan Flow
cytometer (Becton-Dickinson, Mountain View, CA) and CellQuest software
(Becton-Dickinson). In our patient cohort, CD38 expression was
evaluated as a classical diagnostic marker. Considering the CD38 antigenic
expression, the patients were classified into two groups: those
with > or = 20% were considered positive (CD38 + ) and those with <
20% were considered negative (CD38-). Our study focused on the
expression of CD62L on these two groups. Results. CD38 was expressed
in 20% or more of leukemic cells in 17 patients (42,5%), while 23
patients (57,5%) were negative for CD38. The over-expression of CD62L
was detected on 17 patients (74%) who were negative for CD38. Six
patients (26%) of this group had low expression of CD62L.On the other
hand patients who were positive for CD38 had absolute lack of
expression of CD62L (100%). Patients with CD38 + samples have significantly
aggressive disease regardless of their clinical stage. The group of
the patients with expression of CD62L has good clinical progress by far.
Conclusions. The over-expression of CD62L is usually associated with
the absence of CD38 and represented the immunophenotypic signature
of good prognosis in B-CLL. CD62L is an adhesion molecule, which is
involved in the cross-talk between B-lymphocytes with neighboring
endothelial and/or T-cells within the lymph node microenvironment.
CD62L may be used as a diagnostic/prognostic marker for B-CLL, but
more studies are necessary
1310
AQUAGENIC PRURITUS IN POLYCYTHEMIA VERA (PV): HOW IT INFULENCES QUALITY
OF LIFE AND HOW IT CAN BE TREATED: FIRST RESULTS OF A WRITTEN SURVEY OF 123
PATIENTS IN GERMANY
F.P. Siegel, P.E. Petrides
Hematology Oncology Center Munich, MUNICH, Germany
Background. Aquagenic pruritus (AP) is a debilitating condition occuring
in patients with PV. It is characterised by strong itching or stinging
following contact with water without visible changes of the skin. Treatment
of this condition is difficult, but very important as it affects quality
of life in affected patients and is responsible for most of the suffering
in polycythemia vera. Very little is known about the frequency of this
symptom, its precise description, its influence on the quality of life and
its optimal treatment. Aims. For these reasons we intended to document
haematologica/the hematology journal | 2007; 92(s1) | 473