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

12 th Congress of the European Hematology Association
compared the values to15 control males at the same age. Results. 1. In all
group we found lower values of inhibin B than in control (116.85±107.71
ng/mL vs. 196.53 ng/mL±66.8, p=0.02). The patients treated before
puberty also presented low levels of inhibin B (111.72±77.26 ng/mL).
The values of testosterone, FSH and LH were normal. 2. In patients
treated for ALL, the values of inhibin B were lower (91.39±77.26 ng/mL,
p=0.0002) than in control and lower than in patients treated for NHL
(157.17±137.77 ng/mL). The lowest inhibin B values were found in
patients received radiotherapy for CNS (87.26±37.88 ng/mL, p=0.0001).
3. The values of inhibin B lower than 2 SD were observed in 10 patients
treated for ALL and 5 patients treated for NHL. Nine were treated before
puberty. Conclusions. Antileukemic treatment disturbs germinal cell function
without influence on steroidogenesis. Prepubertal state does not
protect the gonads from their failure by anticancer treatment.
1103
CD39 EXPRESSION IN T-LYMPHOCYTES IS ABNORMAL IN CHRONIC LYMPHOCYTIC
LEUKEMIA
D. Pulte, 1 M. Broekman, 2 J. Kizer, 2 R. Furman, 2 A. Marcus1 1 VA NY Harbor Healthcare System, NEW YORK, USA; 2 Weill Medical College
of Cornell Univ, NEW YORK, USA
Background/Aims. Alterations in the T-lymphocyte population in
patients with the B-lymphocyte malignancy chronic lymphocytic
leukemia (CLL) have been identified. Early studies noted an increase in
the CD8 + subpopulation, leading to an abnormal CD4:CD8 ratio. More
recent studies have demonstrated an abnormal response to antigens,
increased CD45RO + cells, and a variety of alterations in CD25 expression
in the T-lymphocytes of CLL patients. CD39 (NTDPase-1) is a cell
surface molecule that mediates platelet reactivity and immune function
via metabolism of ATP and ADP. CD39 is observed on the majority of
B-lymphocytes and a T-lymphocyte sub-population, including subsets
of both CD4 + and CD8 + cells. CD39 expression is associated with activation
and memory cells in T-lymphocytes. We previously demonstrated
that CD39 is present and active on the neoplastic cells in CLL. Our
observations suggested that CD39 activity on B-lymphocytes decreases
as the disease progresses. We now report on changes in CD39 expression
on T-lymphocytes in CLL patients. Methods. Whole blood from
patients with CLL and controls was stained with antibodies against
CD3, CD4, CD8, CD19, CD25, CD39, and CD45RO and incubated for
1 hour in dark with shaking. Samples were lysed with FACS lysis solution
and washed twice with phosphate buffered sodium (PBS). Cells
were resuspended in 0.5 mL PBS and analyzed on a FACS Canto using
FACS Diva software. Statistics were derived using Student’s 2-way Ttest
with unequal variances. Results. Blood samples from 25 patients
with CLL and 23 controls were analyzed. CD39 expression was
increased on T-lymphocytes from patients with CLL as compared to
controls in all T-lymphocytes (Table 1), in both the CD4 + and CD8 + population
(Table 1).
Table 1. CD39 positivity as % all cells of given sub-type.
Subgroup analysis of CLL patients showed that T-lymphocyte CD39
expression was higher in patients with Rai stage III-IV vs 0-II disease
(13.8% vs 28.2%, p=0.05) and those who had received chemotherapy
406 | haematologica/the hematology journal | 2007; 92(s1)
(12% vs 27%, p=0.01). We found a trend toward decreased CD25
expression in CLL patients versus controls; however, the differences
were not statistically significant. The CLL patients had fewer
CD3/CD25/CD39 triple positives compared with controls (18% vs
42%, p=0.006). Conclusions. T-lymphocyte CD39 expression is increased
in patients with CLL. In contrast to B-lymphocytes, increased T-lymphocyte
CD39 expression is associated with more advanced disease in CLL.
CD39 is associated with activation and memory cells; therefore, the
increase in CD39 seen in patients may be due to an increase in memory
T-lymphocytes. However, CD39 expression was seen in some CD25-
/CD45RO- T-lymphocytes in patients, where it is rarely observed in
controls, suggesting that some of the CD39 expression seen in T-lymphocytes
may be aberrant. It has been reported that secretion of IL-2
requires extracellular ATP. High CD39 expression may decrease extracellular
ATP thereby decreasing IL-2 secretion from T-lymphocytes. This
could explain the previously reported observation that IL-2 secretion is
decreased in a mixed lymphocyte reaction of CLL T-lymphocytes and
the CD25 aberrations seen in CLL patients. These findings provide further
support for the thesis that T-lymphocyte abnormalities are involved
in the pathogenesis of CLL and suggest further avenues of research into
the etiology of CLL.
1104
DIFFERENT METHODS FOR ESTIMATION OF ANGIOGENESIS IN PATIENTS
WITH MULTIPLE MYELOMA
O. Markovic, 1 D. Marisavljevic, 1 V. Cemerikic, 2 M. Perunicic, 3
A. Vidovic, 3 M. Bakrac, 3 I. Elezovic, 3 M. Colovic3 1 KBC Bezanijska kosa, BELGRADE, Serbia; 2 Histolab, BELGRADE; 3 Institute
of Hematology, KCS, BELGRADE, Serbia
Background. Angiogenesis is an essential component in the growth
and progression of myeloma plasma cells. Different methods were used
to estimate the intensity of angiogenesis in patients with multiple
myeloma (MM). Aims. The purpose of the study was to establish the
intensity of angiogenesis in patients with MM and to compare two
methods for angiogenesis estimation. In addition, clinical significance of
angiogenesis in patients with MM was particularly analyzed. Patients and
Methods. We analyzed bone marrow biopsy specimens obtained from
59 patients with de novo MM. The clinical staging was done according
to the Durie and Salmon classification (four patients had disease stage
I, 16 patients stage II and 39 patients stage III). Bone marrow angiogenesis
was analyzed using standard imunohistochemical analysis of B5fixed
and routinely processed, paraffin-embedded bone marrow specimens
with antibody against CD34. Bone marrow angiogenesis was estimated
by two different methods. Microvessel density (MVD) was estimated
by counting number of microvessels in three hot spots at magnification
x400, according to the method of Weidner et al. (N Engl J Med
1991; 324:1-8). Semiquantitative estimation of angiogenesis was based
on visual assessment of slides at magnification x100, according to the
method of Rajkumar et al. (Clin Canc Res 2000; 6:3111-16). Each slide
was assigned as low, intermediate or high angiogenesis intensity. Results.
Median MVD was 15 (range: 1-89). Intensity of angiogenesis estimated
by semiquantitative method was assigned as low in 24 (40.67%)
patients, intermediate in 17 (28.81%) patients and high in 18 (30.50%)
patients. There was a statistically significant association between MVD
and semiquantitatively estimated intensity of angiogenesis (p