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

0217
TUBERCULOSIS AMONG A COHORT OF 35 PATIENTS WITH BLOOD DISEASES
C. Ulibarrena, 1 E. Lavilla, 2 J.L. Sastre 1
1 Complexo Hospitalario de Ourense, OURENSE; 2 Complexo Hospitalario Xer-
al-Calde, LUGO, Spain
Tuberculosis (TBC) is still a common infection in Spain, especially in
the North-Western region of Galicia, with Incidence/prevalence rates
over 40 cases /100,000 inhabitants. Tuberculous infection in hematological
patients has been scarcely studied. Methods.We describe the pattern
of infection in 35 patients from 3 Hospitals in Galicia. Data were
obtained from a questionnaire sent to Hematology Services, which gathered
cases form their databases, necropsic reports and reports from
Microbiology Services. The diagnostic criteria for TBC were those
defined by WHO (010 to 018 in CIE-9 classification); infections needed
to be documented with microbiological and/or pathological data. 'Blood
diseases' included acute leukemias, myeloproliferative disorders (MPD),
myelodysplastic syndromes (MDS), myeloma (MM), lymphoma and
aplastic anemia. The diagnosis or TBC should have been made concomitant
or subsequent to the hemopathy, not priorly. Results. 35 HIV (-)
patients were included (median age, 73.7; range 53-86): 15 non-Hodgkin
lymphoma; 6 MDS; 5 MPD (3 essential thrombocythemia; 2 CML); 3
MM; 3 acute leukaemia; 3 chronic lymphocytic leukaemia. The involved
organs were: lymph nodes, 15 (41%); lung, 14 (35%); kidney, 1; skin, 1;
peritoneum, 1; bone, 1; pericardium, 1; 3 cases presented with the miliary
form. Diagnosis was obtained by: detection of acid-fast bacilli, 12
cases; culture, 24; examination of tissue, 18 (2 fine-needle aspiration, 16
biopsies). The majority of biopsies came from adenopathies (neck, 10;
axilar, 2; mediastinum, 1; 2 unknown). 26 patients have died when data
were collected. Post-mortem diagnosis was made in 5 cases (4 lung; 1
peritoneal); the infection could have contributed to the death of other 13.
In 12 cases, the diagnosis of TBC and the hematologic disease was concomitant
(lung, lymph nodes); in the other 23, TBC was found after the
other disease (median interval, 31.7 months; range: 1-90 mo.). In 3
patients there was a history of probable TBC more than 30 yr. before.
Only 12 patients had recorded data of PPD test: it was negative in 4; due
to several reasons, positivity was followed by prophylaxis with isoniazid
only in 1of the remaining 8 (who developed TBC anyway 9 yr. after).
According to IDSA criteria, former risk procedures were: glucocorticoids,
19 cases; chemotherapy (including low dose araC), 15; in 11, no adscription
to risk groups could be established; 4 received only hydroxyurea.
29 patients diagnosed while being alive received anti-TBC therapy. Conclusions.
TBC may be extremely proteiform in hematologic patients
(especially in the elderly), with a majority of extrapulmonary cases,
affecting unusual areas (lymph nodes predominantly). The infection may
occur concomitantly or after the start the hemopathy, in both cases posing
a difficult diagnostic problem due to the coincidence of symptoms
with blood diseases. In highly prevalent areas like ours, TBC must be
always taken into account, even in patients not belonging to risk groups,
since the delay of therapy may be lethal. PPD test should also routinely
performed in hematologic patients, even though its negativity does not
preclude latent infection and prophylaxis is not completely protective nor
safe.
12 th Congress of the European Hematology Association
Myelodysplastic syndromes I
0218
PRION-LIKE DOPPEL GENE (PRND): A NEW MOLECULAR MARKER POTENTIALLY
INVOLVED IN LEUKEMOGENESIS
R. Invernizzi, 1 E. Travaglino, 1 C. Benatti, 1 A. Azzalin, 2 B. Rovati, 1
S. Comincini2 1 2 Fondazione IRCCS Policlinico S. Matteo, PAVIA; University of Pavia, PAVIA,
Italy
The PRND gene encodes Doppel (Dpl), a protein that is strongly
expressed in testis and at much lower levels in other tissues. Despite the
recent discovery of Dpl involvement in spermiogenesis and in apoptotic
death of cerebellar neurons, the physiological role of this prion-like
protein remains unknown. Recently, we observed a weak Dpl expression
in normal CD34 + bone marrow cells, whereas high levels of PRND
transcripts were detected in leukemic cell lines as well as in bone marrow
cells from patients with acute myeloid leukemia (AML) or
myelodysplastic syndrome (MDS). In order to clarify the clinical and
biological relevance of Dpl overexpression in these disorders, we
searched for possible correlations among Dpl expression, some biological
parameters and clinical-pathological features. In some MDS patients
we sequentially studied Dpl levels, to evaluate their changes in the time
as well as the influence of the disease progression and of therapy, whereas
in AML patients treated with aggressive polychemotherapy Dpl levels
were sequentially quantified to assess minimal residual disease.
Moreover, we characterized PRND transcriptional and translational patterns.
Immunocytochemistry, flow cytometry, biochemical and molecular
(real-time quantitative PCR) studies were carried out on bone marrow
cells from 64 AML patients at diagnosis, after induction therapy
and at relapse, and from 98 MDS patients at diagnosis and during disease
progression. Controls were 16 non-hemopathic subjects. Dpl, barely
detectable in normal controls, was detected in almost all AML and
MDS cases, with median percentages of positive cells of 11.5% (IQR 7-
24%), and 17.5% (IQR 11-29%) respectively. In AML no significant relationship
was observed between Dpl levels and clinical and laboratory
features nor did Dpl levels predict response to therapy. In patients achieving
complete remission (22/34) a significant reduction of both transcript
and protein levels (p=0.02) was observed. In 5 relapsing patients Dpl
was again overexpressed at levels similar to those observed at onset.
Also in MDS, Dpl levels were unrelated to clinical and laboratory aspects
nor did they predict disease progression. Their behaviour was variable
during evolution towards acute leukemia. In pathological samples Dpl
was abnormally localized in the cell cytoplasm, while normal CD34+
cells exhibited the expected membrane localization. The ectopic localization
was probably dependent on abnormal cellular trafficking because
of glycosylation pattern modifications of the protein. Moreover, in
pathological samples an abnormal nuclear retention of the transcript was
observed. In conclusion, our findings confirm the clinical usefulness of
Dpl evaluation for AML or MDS diagnosis and for the assessment of
minimal residual disease. Moreover, they suggest its possible physiopathological
role at least in the early phases of cell transformation.
Studies are in progress to better understand which factors may contribute
to the modulation of PRND activity: identifying the promoter
region and critical elements for the activation of the gene may provide
new insights into the involvement of Dpl in leukemic transformation.
0219
BIOLOGICAL AND CLINICAL RELEVANCE OF MATRIX METALLOPROTEINASES 2 AND 9 IN
ACUTE MYELOID LEUKEMIAS AND MYELODYSPLASTIC SYNDROMES
R. Invernizzi, E. Travaglino, C. Benatti, L. Malcovati, M.G. Della Porta,
M. Cazzola
Fondazione IRCCS Policlinico S. Matteo, PAVIA, Italy
Matrix metalloproteinases (MMP) are a family of zinc-dependent
endopeptidases which are able to degrade all the protein components of
the extracellular matrix. MMP-2 (type IV collagenase, gelatinase A) and
MMP-9 (type V collagenase, gelatinase B) have been implicated in tumor
progression and metastasis and, recently, it was suggested that these
enzymes may also contribute to leukemic dissemination. We analyzed
the expression of MMP-2 and MMP-9 in bone marrow cells from 54
patients with acute myeloid leukemia (AML), 153 patients with
myelodysplastic syndrome (MDS) (68 RA, 32 RARS, 31 RAEB, 5 RAEBt,
17 CMML), not previously treated, and 52 non hemopathic subjects,
haematologica/the hematology journal | 2007; 92(s1) | 79