ALBERT

Description: Host responses controlling blood-stage malaria include both innate and acquired immune effector mechanisms. During Plasmodium chabaudi infection in mice, a population of CD11bhighLy6C+ monocytes are generated in bone marrow, most of which depend on the chemokine receptor CCR2 for migration from bone marrow to the spleen. In the absence of this receptor mice harbor higher parasitemias. Most importantly, splenic CD11bhighLy6C+ cells from P chabaudi–infected wild-type mice significantly reduce acute-stage parasitemia in CCR2−/− mice. The CD11bhighLy6C+ cells in this malaria infection display effector functions such as production of inducible nitric oxide synthase and reactive oxygen intermediates, and phagocytose P chabaudi parasites in vitro, and in a proportion of the cells, in vivo in the spleen, suggesting possible mechanisms of parasite killing. In contrast to monocyte-derived dendritic cells, CD11bhighLy6C+ cells isolated from malaria-infected mice express low levels of major histocompatibility complex II and have limited ability to present the P chabaudi antigen, merozoite surface protein-1, to specific T-cell receptor transgenic CD4 T cells and fail to activate these T cells. We propose that these monocytes, which are rapidly produced in the bone marrow as part of the early defense mechanism against invading pathogens, are important for controlling blood-stage malaria parasites.

Description: Abstract 6 Acute promyelocytic leukemia (APL) is a curable disease, and contemporary treatment based on the combination of all-trans retinoic acid (ATRA) with anthracyclines results in overall survival (OS) rates of around 90% at five years. Unfortunately, the treatment outcome of patients with APL in developing countries is significantly less. A recent Brazilian study had reported an OS of 53% with a first 5-days mortality of 13.4%. The International Consortium on Acute Promyelocytic Leukemia (IC-APL) is an initiative of the International Members Committee of the ASH and the project aims to reduce this gap through the establishment of international network, which was launched in Brazil, México and Uruguay. All patients with a suspected diagnosis of APL were immediately started on ATRA, while bone marrow samples were shipped to a national central lab where genetic verification of the diagnosis was performed. Results of the immunofluorescence for PML was obtained within hours and upon confirmation of the diagnosis, patients were enrolled in a protocol identical to the PETHEMA-LPA 2005, except for the replacement of idarubicin by daunorubicin. Supportive care aimed at maintaining platelet counts above 30,000/μl and fibrinogen levels above 150 mg/dl. In each country, cases were discussed every other week through internet and whenever needed international experts were involved. As of June 2009, 102 (70 Brazil, 25 Mexico, 7 Uruguay) APL patients were enrolled. The median age was 34 y (range: 9–72y) with 55 males (54%).The median white blood cell counts (WBC) at baseline was 3.6×109 /L(range: 0.2–149.7). The distribution of the relapse risk score at diagnosis according to PETHEMA-GIMEMA criteria was 14 low (14%), 54 intermediate (53%) and 34 high risk(33%) respectively. The incidence of low risk APL appeared lower than the values reported in developed countries. Of 102, 97 patients have toxicity and response data available. Of these 97, 12 (12.3%)experienced at least three symptoms/signs of differentiation syndrome (DS) and 77 (79%) patients achieved a complete remission (CR). Twenty-three deaths occurred and the cause of deaths included 9 hemorrhage, 8 infection, 2 DS . The 7 and 30 day mortality rates were 8% and 19.6%, respectively, and the one- year overall survival was 75% (95%CI:68%–84%). The median follow-up time among survivors was 14 months (range: 1.3–35). Among 77 patients who achieved CR, the 1-year OS and disease-free survival from the date of CR was 95% (95% CI: 89%–100%). Only one patient relapsed. For patients surviving a minimum of 30 days the outcome was similar to that reported by the twin PETHEMA-LPA 2005 protocol in European patients. Prognostic factors for overall survival were examined using log-rank test as well as multivariate Cox models. Factors predicting OS were a high relapse risk score at baseline (1-year OS: 59% for high, 87% for intermediate, 91% for low, p=0.0007) and age. The 1-year OS was 85% for age

Description: Several authors have reported ASCT as a feasible, safe and effective treatment in HIV associated lymphoma patients (pts) receiving Highly Active Antiretroviral Therapy (HAART), particularly when being autografted with chemosensitive disease. To gain a better understanding of the usefulness of ASCT in HIV+ Lymphoma pts a retrospective comparative study (HIV+ vs HIV- lymphoma pts with ASCT) has been performed using the EBMT-LWP Registry. Methodology: Registry-based, retrospective, individually matched case-control analysis. Within the participating centres one or two HIV- controls for each HIV+ pt were selected from the registry with the following inclusion criteria: Known HIV serological status at ASCT, lymphoma (HL or NHL), ASCT performed between 1999 and 2006 and pts over 18 years of age. Two cohorts (HIV+ vs HIV-) were matched for histology, IPI at diagnosis (NHL), stage at diagnosis, disease status at ASCT, age at ASCT, year and country of ASCT. Results: 40 HIV+ lymphoma pts undergoing an ASCT were matched with 46 HIV- pts. Pts and transplant features are shown in table 1. With a median follow up of 36 mo, the differences regarding OS and PFS were not significant: 62% for HIV+ vs 69% for controls, and 56.5 vs 58%, respectively. No differences were seen regarding HL and NHL pts. An overall TRM of 10% was observed in the HIV cohort, mainly related to infections, while no cases of TRM were seen within the control arm. Since survival rates between the HIV+ and the matched HIV- lymphoma populations remained comparable, the HIV condition should not preclude these pts from being treated according to the same criteria as the HIV negative lymphoma population. Nevertheless, infectious complications should be cautiously followed within the HIV+ lymphoma pts undergoing an ASCT. Patients and transplant features HIV+ HIV- n=40 % n=46 % Age [Median (range)] in years 41.4 (29.2–62.5) 44 (16–62.4) p= NS Male sex 35 87.5 25 54.3 p=0.001 Histology     DLBCL 24 60 25 54.3 p= NS     Burkitt lymphoma 2 5 2 4.3 p= NS     T-cell NHL 2 5 3 6.6 p= NS     HL 12 30 16 34.8 p= NS Disease status at ASCT     Complete remission (CR) 21 (12 in CR1) 52.5 (30 in CR1) 20 (11 in CR1) 43.5 (24 in CR1) p= NS     Chemosensitive disease 16 40 25 54.3 p= NS     Chemorefractory disease 3 7.5 1 2.2 p= NS CD34+ cells infused mill/kg [median (range)] 4.9 (1.6–21.2) 4.8 (0.9–21.2) p= NS     G-CSF prior to engraftment 36 90 21 46 p= 0.0001     Neutrophil engraftment 39 98% 46 100 p= NS Cause of Death     Relapse/progression 9 60% 10 84% p=0.08     Secondary malignancy 1 6.7% 1 8% p= NS     Transplant-related deaths 4 26.7% 0 0% p=0.06     Other 1 6.7% 1 8% p= NS

Description: Background: Autologous Transplantation with intermediate or high doses of Melphalan is the most effective therapy in patients with Multiple Myeloma being 〈 65 years old, although this procedure is limited by an high percentage of relapses. Recent advances knowledge of the molecular mechanisms, allowed the development of novel targeted therapies. Several studies have shown that the synergic effect of Bortezomib and Talidomidomide, in association with conventional chemotherapies, induces a more pronounced reduction of tumour mass, an enhancement of global responses and an improvement of Progression Free Survival (PFS). Aims: The present study is aimed at the evaluation of effectiveness and toxicity of MVTD conditioning therapy with stem cell support in Multiple Myeloma relapsed patients. Patients and Methods: From January 2005 to August 2007, 15 patients (7 males and 8 females: median age 62 yrs (46–70)) have been subjected to Autologous Transplantation therapy with MVTD conditioning therapy (Melphalan 100 mg/sqm days - 6 and - 3, Bortezomib 1.3 mg/sqm days - 6 and - 3, Talidomide 200 mg from day - 6 to day - 2, Desametazone from day - 6 to day - 3, CSP infusion at day 0). All patients were in CS III A; median performed therapies 3 (1–5); all had received previously a therapy with Talidomide, two Talidomide + Velcade, one PEG-IFN. Results: 100% of patients obtained a response: 4 patients (26.7%) obtained CR, 5 (33.3%) a nCR, 4 (26.7%) a VGPR and 2 (13.3%) a PR. One patient in CR was subjected to 2 therapeutic lines and 3 patients in CR were subjected to more than 1 therapeutic lines, 5 in nCR from 2 to 4, 4 in VGPR from 2 to 4 and 2 in PR more than 2. At the moment, with a median of 7.5 months (1–21), 11 (73%) are alive, 8 (53%) still keep the obtained response, 3 (20%) after progression are in rescue therapy, 4 patients (27%) died because of progression. PFS median since MVTD beginning is of 6 months (1–24). None of the patients have developed a significant neurotoxicity. All patients have developed grade 4 thrombocytopenia and neutropoenia, with a median of 3 days (1–10) and 6 days (4–12), respectively. 54% of patients had neutropoenia with fever with a median of 2 days (0–8). The median units of platelets and red cells infused was 3 (2–11) and 2 (0–11), respectively. Conclusions: In our experience, Autologous Transplantation therapy with MVTD conditioning, appeared to be effective in relapsed patients with Multiple Myeloma. Although in a small series of patients the response obtained was independent from the number and type of previous therapies, from the clinical state, and β2 microglobulin. No significant neurotoxicity has been observed, whether the haematological toxicity was overlapping to that due to conventional Autologous Transplant. Further studies, in larger series of non pluritreated patients are required.

Description: In paroxysmal nocturnal hemoglobinuria (PNH) hemolytic anemia is due mainly to deficiency of the complement regulator CD59 on the surface of red blood cells (RBCs). Eculizumab, an antibody that targets complement fraction 5 (C5), has proven highly effective in abolishing complement-mediated intravascular hemolysis in PNH; however, the hematologic benefit varies considerably among patients. In the aim to understand the basis for this variable response, we have investigated by flow cytometry the binding of complement fraction 3 (C3) on RBCs from PNH patients before and during eculizumab treatment. There was no evidence of C3 on RBCs of untreated PNH patients; by contrast, in all patients on eculizumab (n = 41) a substantial fraction of RBCs had C3 bound on their surface, and this was entirely restricted to RBCs with the PNH phenotype (CD59−). The proportion of C3+ RBCs correlated significantly with the reticulocyte count and with the hematologic response to eculizumab. In 3 patients in whom 51Cr labeling of RBCs was carried out while on eculizumab, we have demonstrated reduced RBC half-life in vivo, with excess 51Cr uptake in spleen and in liver. Binding of C3 by PNH RBCs may constitute an additional disease mechanism in PNH, strongly enhanced by eculizumab treatment and producing a variable degree of extravascular hemolysis.

Description: Autologous stem cell transplantation (ASCT) has been successfully used in HIV-related lymphoma (HIV-Ly) patients on highly active antiretroviral therapy. We report the first comparative analysis between HIV-Ly and a matched cohort of HIV− lymphoma patients. This retrospective European Group for Blood and Marrow Transplantation study included 53 patients (66% non-Hodgkin and 34% Hodgkin lymphoma) within each cohort. Both groups were comparable except for the higher proportion of males, mixed-cellularity Hodgkin lymphoma and patients receiving granulocyte colony-stimulating factor before engraftment and a smaller proportion receiving total body irradiation-based conditioning within the HIV-Ly cohort. Incidence of relapse, overall survival, and progression-free survival were similar in both cohorts. A higher nonrelapse mortality within the first year after ASCT was observed in the HIV-Ly group (8% vs 2%), predominantly because of early bacterial infections, although this was not statistically significant and did not influence survival. Thus, within the highly active antiretroviral therapy era, HIV patients should be considered for ASCT according to the same criteria adopted for HIV− lymphoma patients.

Description: ASCT has been reported as a feasible, safe and effective treatment in HIV-Ly patients (pts) receiving Highly Active Antiretroviral Therapy (HAART). We hereby present an updated analysis of the EBMT experience on HIV-Ly pts treated with an ASCT since 1999. Sixty eight pts from 20 institutions [56 (82%) males, median age of 41 (29–62) years] were included. Twenty-two pts met AIDS criteria (other than lymphoma) at the time of HIV-Ly diagnosis. Forty-nine pts were diagnosed of NHL (31 DLBCL; 8 Burkitt/Burkitt-like; 4 plasmablastic; 3 anaplastic, 3 PTCL), 80% of them presenting with stage 〉II and 18 pts of HL, 61% of them presenting with stage 〉II. Median (range) lines of therapy before ASCT was 2 (1–5). Thirty-five pts were autografted in CR (16 in CR1), 25 in chemosensitive disease and 8 in chemoresistant disease. Sixty-five pts received the BEAM protocol as a conditioning regimen and the remaining three received TBI-based protocols. Two pts received more than 1 ASCT (censored at time of 2nd ASCT). At the time of ASCT the median number of CD4+ cells was 162 (8–1159)/mcl and 34 pts had undetectable HIV viral loads. HAART was given in 55/57 pts during conditioning but withdrawn in 25% of them. The median number of CD34+ cells infused was 4.5 (1.6–21.2) ×106/kg and G-CSF was used until engraftment in 60/67 pts for a median of 8 (2–21) days. All pts but one who died on day +15 reached neutrophils〉500/ml at a median time of 11 (8–36) days. Platelet count 〉20.000/ml was reached in 61 pts at a median time of 14 (6–455) days. Twenty three pts (34%) died: disease-progression (n=15), acute ASCT-related complications (n=6) [bacterial infections (n=4), multi-organ failure (n=1), other complications (n=1)] and 2 pts died from HIV-related complications. Cumulative incidence of NRM was 4.4% (95%CI 1.5–13.3) and 7.6% (95%CI 3.3–17.6) at 3 and 12 months, respectively. Age 〉 50 years at ASCT [RR 4.37 (95%CI 1.01–18.89), p = 0.05] was the only independent adverse prognostic factor for NRM. Relapse occurred in 19 (28%) pts giving a cumulative incidence of 23.6% (15.2–36.9) and 29.6% (20.0–43.8) at 12 and 24 months, respectively. Median time to progression was 4.5 (0.5–32) months. Histology (NHL other than DLBCL) [RR 3.2 (95%CI 1.0–9.6), p = 0.04], the use of 〉2 previous treatment lines [RR 2.7 (95%CI 1.0–7.2), p = 0.05] and not being in CR at ASCT [RR 3.5 (95%CI 1.2–9.7), p = 0.02] were significantly associated with a higher risk of relapse post-ASCT. With a median follow up time of 32 (2–81) months, PFS and OS were 56% (CI95% 43–70) and 61% (CI95% 48–74) at 3 years, respectively. Pts with refractory disease showed a poorer OS [RR 5.1 (95%CI 1.8–14.6), p = 0.002] and PFS [RR 5.3 (95%CI 2.0–13.7), p = 0.001]. One pt developed an in-situ epithelioma and myelodisplastic syndrome (+4y) and another one a kidney adenocarcinoma (+3y). The results of the largest experience on ASCT for HIV-Ly indicate that this approach is a useful treatment in terms of NRM, long-term OS, and PFS, with significantly better results in patients autografted with chemosensitive disease.

Description: Abstract 1894 Poster Board I-917 Background Chronic Mountain Sickness (CMS) is a clinical entity that occurs in native or long-life residents above 2500 meters of altitude. The disease is characterized by massive erythrocytosis, hyperviscosity syndrome (headache, dyspnea and cyanosis) severe hypoxemia and cardiopulmonary symptoms. The etiology is unknown and no association has been found with Erythropoietin (EPO), Epo receptor (EpoR), Hypoxia Inducible Factor 1a (HIF-1a), von Hippel Lindau (VHL), as well as PHD1, PHD2, PHD3 or PTEN genes. Therapy relies on phlebotomy and oxygen support. Acetazolamide, Medroxyprogesterone and Enalapril have also been tested, but their use has not been largely implemented. Since HMG-CoA inhibitors such as farnesyltransferase inhibitors (Larghero, Blood 2005) may inhibit the in vitro autonomous erythropoiesis of polycythemia vera patients, we studied in CMS the therapeutic potential of statins that have similar pharmacologic activity. Patients and Methods Normal controls (NC, n= 10) and patients were native Bolivians from the city of La Paz, Bolivia (3600–4000 mt altitude). The diagnosis of CMS (n=15) was made according to the consensus statement on this disease (Leon-Velarde, 2005). The diagnosis of Polycythemia Vera (PV, n= 5) or secondary erythrocytosis (SE, n= 10) was done according to WHO criteria or established clinical guidelines. Serum Erythropoietin (sEpo) was assessed by chemiluminescent assay. Burst forming units-erythroid (BFU-E) assay was performed by plating 105 BM mononuclear cells in methylcellulose with or without recombinant human rhEpo (2IU/ml) and Simvastatin (20 mM). Evaluation of apoptosis by Annexin V/7-AAD and JAK2V617F mutational analyses were performed as described (Guerini et al, Leukemia 2008). Results CMS patients (median age 48 years, range 29–58) had median values of hemoglobin and hematocrit (Hb 20.3 gr/dl, range: 19.1–22 and Hct, 62%) significantly higher than observed in NC (Hb 16.2 gr/dl, range 14.8–16.5 and Htc 52%), respectively (p〈 0.001) and significantly lower than SE patients (Hb: 22.8 gr/dl, range 20.2–25 and Htc: 71%), (p

Description: Abstract 1347 Poster Board I-369 Introduction Sepsis is among the top 10 causes of death, but improvements in the diagnostic tests for detecting and monitoring sepsis and infection have been limited in the last years. Neutrophil CD64 expression increases rapidly in the presence of inflammation mediators and in response to infection and tissue damage. We have evaluated changes in the expression of neutrophil CD64 in infected patients in comparison with other markers of infection and sepsis. Methods Prospective analysis of 56 blood samples from patients from the intensive care unit at our institution was performed for neutrophil CD64 expression, C-reactive protein (CRP), automated absolute neuthophil count (ANC), and complete manual leucocyte formula including % of bands (BANDS), and % of metamyelocytes and myelocytes (IG). Neutrophil CD64 expression was measured by flow cytometry using a quantitative method (Leuko64TM, Trillium Diagnostics, LLC). Patients were categorized into 5 groups (CLINIC) based on the clinical history and the degree of a systemic inflammatory response, from 1 (no inflammation) to 5 (septic shock). Statistics were performed using linear regression, correlation coefficient, and Passing-Bablock (P-B) regression. Sensitivity (S), specificity (SP), efficiency (E), and positive and negative predictive values (PPV and NPV respectively) were analyzed for all the parameters measured. Results Our results showed a correlation with CLINIC of 0.417, 0.552, 0.268, and 0.136 for CD64, CRP, BANDS, and ANC, respectively. P-B regression was only good for CD64, with a slope of 1.03 (0.6-1.4). Percentages (%) of S, SP, E, PPV, and NPV for CD64 were of 81%, 72%, 71%, 46% and 92%, respectively for groups 4 and 5. For CRP, S was of 93% with SP of 20%, E of 38%, PPV of 27%, and NPV of 91%. The remaining parameters showed deficient correlation with CLINIC. Correlations between CD64 and CRP, BANDS, and ANC were of 0.435, 0.342, and 0.01, respectively. Conclusions Neutrophil CD64 expression quantitation provides improved diagnostic detection of infection/sepsis compared with the standard diagnostic tests used in current medical practice. CD64 expression showed a better PPV than CRP, and an acceptable NPV. CRP showed deficient SP and E. BANDS, GI, and ANC showed no correlation with CLINIC. CD64 is a new indicator of infection that deserves consideration to be introduced in the daily hematology laboratory analysis. Disclosures No relevant conflicts of interest to declare.

Description: Acute myeloid leukemia (AML) is a malignant disease characterized by abnormal proliferation of clonal precursor cells. Although different strategies have been adopted to obtain complete remission, the disease actually progresses in about 60–70% of patients. Bortezomib has documented antitumor activity in multiple myeloma and other lymphoid malignancies. TRAIL is a member of the TNF family that induces apoptosis in tumor cells while sparing normal cells. Here we examined the sensitivity to Bortezomib alone or in combination with TRAIL of bone marrow cells from AML patients (34 patients: 25 newly-diagnosed, 4 relapsed, 5 refractory). Immunofluorescence analysis showed that NF-κB was located in the nuclei of the AML blasts and it did not translocate after exposure to Bortezomib. Bortezomib induced cell death in blasts from each patient sample. The cytotoxic effect was dose- and time-dependent (concentration from 0.001 to 10 μM for 24 and 48 hours) and was associated with down-regulation of Bcl-xL and Mcl-1, up-regulation of TRAIL-R1, TRAIL-R2, p21 and activation of executioner caspases. Moreover, low doses of Bortezomib primed TRAIL-resistant AML cells for enhanced TRAIL-mediated killing. Thus, the combination of proteasome inhibitors and TRAIL is effective for treating AML patients, even patients who are refractory to conventional chemotherapy.