Ex Vivo Induction of Multiple Myeloma-specific Immune Responses by Monocyte-derived Dendritic Cells Following Stimulation by Whole-tumor Antigen of Autologous Myeloma Cells.

The introduction of novel agents has significantly expanded treatment options for multiple myeloma (MM), albeit long-term disease control cannot be achieved in the majority of patients. Vaccination with MM antigen-loaded dendritic cells (DCs) represents an alternative strategy that is currently being explored. The aim of this study was to assess the immunogenic potential of ex vivo-generated monocyte-derived DCs (moDCs), following stimulation with the whole-antigen array of autologous myeloma cells (AMC). MoDCs were loaded with antigens of myeloma cells by 2 different methods: phagocytosis of apoptotic bodies from γ-irradiated AMC, or transfection with AMC total RNA by square-wave electroporation. Twenty patients with MM were enrolled in the study. Following stimulation and maturation, moDCs were tested for their capacity to induce T-helper 1 and cytotoxic T lymphocyte responses in vitro. Both strategies were effective in the induction of myeloma-specific cytotoxic T lymphocyte and T-helper 1 cells, as demonstrated by cytotoxicity and ELISpot assays. On the whole, T-cell responses were observed in 18 cases by either method of DC pulsing. We conclude that both whole-tumor antigen approaches are efficient in priming autologous antimyeloma T-cell responses and warrant further study aiming at the development of individualized DC vaccines for MM patients.

BCR-ABL fusion protein detection in peripheral blood and bone marrow samples of adult precursor B-cell acute lymphoblastic leukemia patients using the flow cytometric immunobead assay.

BACKGROUND: The ability to detect the BCR-ABL fusion gene in precursor B-cell acute lymphoblastic leukemia (pB-ALL) is essential for making accurate treatment decisions. METHODS: We used a new flow cytometric immunobead assay for BCR-ABL fusion protein detection in peripheral blood and/or bone marrow samples from 38 adult pB-ALL patients and the results were compared with polymerase chain reaction (PCR) detection of BCR-ABL transcript. RESULTS: The fusion protein was detected in peripheral blood and bone marrow samples from seven of the 38 (18%) patients, and results for both the p190 and p210 were confirmed by PCR. One case, which was positive by cytogenetics and fluorescence in situ hybridization (FISH), was negative by PCR but positive by flow cytometry. Another case, which was positive by PCR and negative by flow cytometry, was from a patient on steroid treatment. CONCLUSIONS: The cytometric immunobead assay for BCR-ABL fusion protein detection was found to be suitable for the investigation of pB-ALL patients. This assay is reliable, rapid and simple to use for peripheral blood and bone marrow samples.

Severity of coronary artery disease in postmenopausal women: association with the androgen receptor gene (CAG)n repeat polymorphism.

OBJECTIVE: Androgen may adversely affect vascular health in women. We investigated the associations between the androgen receptor gene (CAG)n repeat polymorphism, which affects androgen receptor transcriptional activity, and the severity of coronary artery disease (CAD) in women undergoing coronary angiography. METHODS: We examined 131 postmenopausal women (46-82 y). CAD severity was assessed by the number of vessels with greater than 50% stenosis. The history of angina, myocardial infarctions, and biochemical parameters were recorded. CAG repeats ranged between 13 and 24 in the shorter allele and 16 and 28 in the longer allele. The mean lowest quartile corresponded to 19, and the highest, to 22 repeats. RESULTS: Carriers of 19 repeats or less in their shorter allele had severe disease (≥2 vessels affected) and a history of angina more frequently than those carrying 22 or more (39.2% vs 9.5%, P = 0.009 and 80.8% vs 55%, P = 0.037, respectively, using the Fisher exact test). A higher percentage of women carrying 19 repeats or less had one and two myocardial infarctions (28.6% and 10.7%, respectively) compared with women with more than 19 repeats (18.2% and 1.45%, respectively, P = 0.019). Women homozygous for two longer alleles (≥22 repeats) had less severe CAD, significantly higher sex hormone-binding globulin levels, and less frequent antilipid drug therapy compared with those homozygous for shorter alleles (P < 0.05). Total cholesterol and low-density lipoprotein cholesterol levels were negatively correlated with the number of repeats in the shorter allele (P < 0.04). CONCLUSIONS: Shorter polyglutamine stretch in the androgen receptor correlates with more severe CAD and worse predisposing factors in postmenopausal women undergoing coronary angiography. This association may support the adverse cardiovascular effect of lifelong androgenic exposure in this selected group of women.

Acute promyelocytic leukemia: an experience on 95 greek patients treated in the all-trans-retinoic Acid era.

Acute promyelocytic leukemia (APL) is highly curable with the combination of all-transretinoic acid (ATRA) and anthracycline based chemotherapy, but the percentage of early deaths remains high. In the present study, we report the clinical, immunophenotypic, cytogenetic and molecular characteristics and outcome of APL patients diagnosed and treated in various Hospitals of Greece and Cyprus.We describe the data of ninety-five APL patients who were diagnosed during the last 15 years. Seven (7.4%) newly diagnosed APL patients died due to intracranial hemorrhage within 72 hours of presentation. All but two patients were induced with ATRA alone or ATRA plus chemotherapy. The early death rate was 14.9%. After induction all 80 evaluable patients achieved complete hematologic remission. The cumulative incidence of relapse was 18.3%. Eight of the ten relapsed patients were successfully salvaged, while both patients with molecularly resistant disease died during salvage treatment. Overall survival (OS) at 5 years was 78.4% and disease free survival (DFS) 73.6%. In multivariate analysis of OS age over 60 years, DIC at diagnosis and marginally major hemorrhage at presentation were identified as adverse prognostic factors. In the subgroup of patients with available data on FLT3 mutation status (49 out of 94), ITD positivity also remained as an independent prognostic factor in the final model of OS, together with major hemorrhage and marginally high Sanz score. We found a close correlation between the CD2 expression and the development of the differentiation syndrome (DS). In conclusion, the main problem in managing patients with APL is still the high early death rate.

5'RARA submicroscopic deletion from new variant translocation involving chromosomes 15, 17, and 18, in a case of acute promyelocytic leukemia.

Submicroscopic deletions of the PML-RARA fusion genes constitute rare rearrangements in acute promyelocytic leukemia (APL). We describe a rare case of APL carrying a novel complex translocation involving chromosomes 15, 17, and 18 associated with a submicroscopic deletion of the 5' part of the RARA gene, as evidenced by fluorescence in situ hybridization (FISH). A PML/RARA dual-fusion probe did not reveal the RARA-PML fusion signal on the der(17q), usually detected in the typical t(15;17). The RARA break-apart probe showed a deletion hybridization pattern with loss of the signal corresponding to the 5' portion of the RARA gene. Reverse transcriptase-polymerase chain reaction confirmed the absence of the fusion RARA-PML transcript. The patient achieved complete remission, but died during consolidation therapy, 2 months after diagnosis. To our knowledge, this is the first reported case of APL with a complex variant t(15;17) involving chromosome 18 at band q12 and one of the very rare described cases displaying a submicroscopic deletion of the RARA 5' region. Further cases are needed to delineate the incidence of submicroscopic deletions in APL and elucidate their prognostic impact.