ALBERT

Description: Within the broad problem of host immune surveillance versus tumor immune evasion, a most intriguing question is how the cellular immunity can cope with cancerous cells that have gotten rid of the classical antigen-presenting machinery. One such option stems from (1) the fact that HLA loss is often attended with expression of Hsp70 on the tumor cell surface, and (2) our findings that human lymphocytes express a protein Tag7 (also known as PGRP-S) capable of tight and specific interaction with cognate Hsp70. Here we show that a subpopulation of human CD4+CD25+ lymphocytes, obtained either in culture as lymphokine-activated killers or directly from healthy donors, carry Tag7 and FasL on their surface and can indeed kill the HLA-negative tumor-derived cells K562 and MOLT-4 that expose Hsp70 and Fas. The primary binding of lymphocyte Tag7 to target-cell Hsp70 is very specific (eg, it is blocked by preincubating either cell with minimal peptides from the “partner” protein), and secures cell contact indispensable for subsequent FasL/Fas-triggered apoptosis. Unrelated to natural killer cell action or the putative role of Hsp as an antigen-presenting substitute, this novel mechanism is rather a backup analog of orthodox (CD8+) target recognition (Tag7 acting as built-in T-cell receptor and Hsp70 itself as ligand).

Description: The population of new ANHL patients is heterogeneous in terms of their biological parameters and the IPI scoring system is widely used to make the disease prognosis. Recently heterogeneity of ANHL patients in terms of their QoL has been shown. There are four groups of patients with the following grades of QoL impairment: none, mild, significant, and critical impairment. We aimed to study the distribution of new ANHL patients according to the grades of QoL impairment and to identify the relationship between the QoL impairment groups and the IPI groups. 73 new aggressive NHL lymphoma patients (Stage IIB-IV, mean age 55.2 SD=17.6, males/females – 39/34) were involved in this study. QoL assessment was conducted before treatment using SF-36. The method of integral profiles was used to calculate the integral QoL index (IQLI). Patients were stratified using IQLI. Gamma correlation between ranges of QoL impairment groups and IPI groups was used. The following distribution of ANHL patients was demonstrated on the basis of different grades of QoL impairment: 9% - no impairment (IQLI - 0.6); 12% - mild impairment (0.3); 24% - significant impairment (0.13); 55% - critical impairment (0.02). The distribution of patients in these groups who were classified as high-intermediate or high risk according to the IPI was 28%, 67%, 83%, and 90%, respectively. High correlation between ranges of QoL impairment groups and IPI groups was observed: Gamma correlation – 0.6 (p

Description: During the last decade HDCT+ASCT is more often used as a therapeutic option for MS patients. The major treatment outcomes for MS patients are disease-progressive free period and improvement of patient’s quality of life (QoL). We aimed to study treatment outcomes in MS patients after early, conventional and salvage HDCT+ASCT. Thirty two patients with MS (secondary progressive - 16 patients, primary progressive - 8, progressive-relapsing - 1, relapsing-remitting - 7) from 6 medical centers were included in this study (mean age - 32.0, range: 19–49; male/female - 12/20). Two patients underwent early HDCT +ASCT, 27 patients - conventional HDCT+ASCT and 3 patients - salvage HDCT+ASCT. Median EDSS at base-line was 6.0 (range 2.0 – 8.0). The median follow-up duration was 18 months (range 6 – 78 months). All of the patients had previously undergone conventional treatment. Neurological and QoL evaluation was provided at baseline, at discharge, 3, 6, 9, 12 months, and then every 6 months after HDCT+ASCT. MRI was conducted at baseline, at 6, 12 months, and at the end of follow-up. FACT-BMT and FAMS were used for QoL evaluation. QoL response was evaluated using Integral QoL index. All 27 patients with the follow-up longer than 1 year, included in the analysis, experienced a clinical stabilization or improvement. More than half of them improved: 6 patients showed significant improvement in EDSS (by more than 1.0 point), 4 patients improved by 1.0 point, and 5 patients - by 0.5 points on EDSS. Twelve patients achieved stabilization. Two patients deteriorated to a worse score after 18 months of stabilization; 2 other patients progressed after 12 and 30 months of improvement, respectively. All the patients with clinical stabilization and improvement exhibited negative MRI scans. Out of 21 patients included in the analysis of QoL response 19 exhibited improved QoL 6 months post-transplantation. At one year after HDCT+ASCT 1 patient exhibited excellent QoL response; 3 patients - good QoL response; 7 patients - moderate QoL response and 8 patients - minimal QoL response. At 2.5 years post-transplantation 3 more patients had excellent QoL response. Further QoL improvement was observed at longer follow-up. One of the patients who experienced progression after stabilization had no QoL response; another patient who progressed after stabilization had stable QoL during 6 months post-transplant and significantly worsened at 9 months. In conclusion, clinical response was observed in 100% of MS patients after early, conventional and salvages HDCT+ASCT. The majority of patients with clinical response had a good or moderate QoL response. Further studies should be done to investigate the clinical and patient-reported outcomes of HDCT+ASCT in MS patients to better define treatment success.

Description: HDCT+ASCT is a new and promising therapy for MS patients. Among a number of unclear questions is the terms of conducting HDCT+ASCT. According to our concept there are 3 strategies of HDCT+ASCT depending on the terms of disease process: early, conventional and salvage. Another important consideration is that the major treatment outcomes for MS patients are disease-progressive free period and improvement of patient’s quality of life (QoL). With this in mind, evaluation of both clinical and patient-reported outcomes in MS patients after HDCT+ASCT is worthwhile. We aimed to study the clinical and QoL response in MS patients after early, conventional and salvage HDCT +ASCT. 17 patients with MS (secondary progressive − 8 patients, primary progressive − 6, progressive relapsing − 2, relapsing remitting − 1) were included in the study (mean age − 32.3, SD − 6.6; male/female − 4/13). Fifteen patients underwent conventional HDCT +ASCT; one patient (relapsing-remitting MS) - early HDCT+ASCT and one patient (primary progressive MS) - salvage HDCT+ASCT. Median EDSS at base-line was 6.0 (range 2.0 – 7.5). The median follow-up duration was 12 months (range 6 – 72 months). All of the patients had previously undergone conventional treatment. Neurological and QoL evaluation was provided at baseline, at discharge, 3, 6, 9, 12 months, and then every 6 months. MRI was conducted at baseline, at 6, 12 months, and at the end of follow-up. FACT-BMT and FAMS were used for QoL evaluation. QoL response was evaluated using Integral QoL index, which was calculated by the method of integral profiles. 14 (82.4%) out of 17 patients including 12 patients with conventional HDCT+ ASCT, one patient with early HDCT+ ASCT and one patient with salvage HDCT+ ASCT experienced a clinical stabilization or improvement. Three patients showed significant improvement in EDSS (by more than 1.0 point), 2 patients improved by 1.0, and 3 patients - by 0.5. Six cases remained stable. All of the patients with clinical stabilization and improvement exhibited negative MRI scans. One patient continuously worsened and died 3 years after the transplantation. Two other patients worsened by 0.5 points in their EDSS. All patients with clinical response exhibited improved QoL at 6 months post-transplant and they preserved improved QoL at the end of follow-up. At one year after HDCT+ASCT the patients exhibited a good or excellent QoL response. These levels of QoL response were preserved throughout the follow-up period. In conclusion, clinical response was observed in 82.4% MS patients after HDCT+ASCT. Notably, patients undergoing early, conventional and salvage HDCT+ASCT exhibited clinical response. All patients with clinical response had good or excellent QoL response. Thus, HDCT+ASCT appears to be an effective treatment for MS both in terms of clinical and patient-reported outcomes. The data obtained point to feasibility of early, conventional and salvage HDCT+ASCT in MS patients. Further studies should be done to investigate clinical and QoL response in MS patients receiving early, conventional and salvage HDCT+ASCT to better define treatment success.

Description: Unique and shared cytogenetic abnormalities have been documented for marginal zone lymphomas (MZLs) arising at different sites. Recently, homozygous deletions of the chromosomal band 6q23, involving the tumor necrosis factor alpha–induced protein 3 (TNFAIP3, A20) gene, a negative regulator of NF-κB, were described in ocular adnexal MZL, suggesting a role for A20 as a tumor suppressor in this disease. Here, we investigated inactivation of A20 by DNA mutations or deletions in a panel of extranodal MZL (EMZL), nodal MZL (NMZL), and splenic MZL (SMZL). Inactivating mutations encoding truncated A20 proteins were identified in 6 (19%) of 32 MZLs, including 2 (18%) of 11 EMZLs, 3 (33%) of 9 NMZLs, and 1 (8%) of 12 SMZLs. Two additional unmutated nonsplenic MZLs also showed monoallelic or biallelic A20 deletions by fluorescent in situ hybridization (FISH) and/or SNP-arrays. Thus, A20 inactivation by either somatic mutation and/or deletion represents a common genetic aberration across all MZL subtypes, which may contribute to lymphomagenesis by inducing constitutive NF-κB activation.

Description: Introduction: HSCT has been developed in few Wm cases and is nowadays challenged by other innovative approaches. However, high dose therapy followed by autologous HSCT (HD-auto) produces high response rate and some long term responses while allogeneic HSCT performed after either myeloablative (MA-allo) or reduced intensity conditioning (RIC-allo) regimens may be cure of Wm (Dreger 1998, Tournilhac 2003, Maloney 2006). Methods: We updated and extended our retrospective experience on 32 HD-auto, 11 MA-allo and 11-RIC-allo performed from 1990 to 2006 in 51 patients from 18 institutions. A MA-allo and a RIC-allo were performed in 1 and 2 cases respectively following relapse after a 1st HD-auto. Results: Data are presented in the table. HD-auto MA-allo RIC-allo Nb 32 11 11 Median age at transplant 56 46 56 Interval: diagnosis-transplant 38 50 74 Chemoresistance at transplant 25% 36% 55% Conditioning regimen BEAM (13), TBI/melphalan (9), TBI/endoxan (7), other (3) TBI/endoxan (9), other (2) TBI/fluda (10), other (1) Donor Sibling (9), unrelated (2) Sibling (8), unrelated (2), cord blood (1) Median follow up (m) 45 (3–121) 68 (3–132) 22(2–60) Relapse 18 (56%) 4 (36%) 0 Transplant related mortality 12,5% (one 2th cancer) 36% 27% (one 2th cancer) overall survival (1;3;5y) 87%, 77%, 58% 64%, 54%, 54% 82%, 68%, 68% Event free survival (5y) 25% 48% 68% Acute GVHD developed following 9 MA-allo [Grade III-IV (n=1)] and 8 RIC-allo [Grade III-IV (n=1)]. Chronic GVHD developed following 7 MA-allo [limited (n=5), extensive (n=2)] and 5 RIC-allo [limited (n=2), extensive (n=3)]. Conclusion: We confirm that autologous HSCT achieves some long term responses even in heavily pretreated patients. Allogeneic HSCT induces very long term disease control and may cure WM. Specially, the RIC-allo gives impressive results on disease control in a set of older patients, with refractory disease, mostly heavily pretreated.

Description: Thiopurine S-methyltransferase (TPMT) is a key enzyme in the catabolism of 6-mercaptopurine (6MP), a key component of therapy for childhood acute lymphoblastic and promyelocytic leukemias and lymphoblastic non-Hodgkin lymphoma. TPMT mutations reduce TPMT enzymatic activity; therefore, TPMT-deficient patients develop severe hematopoietic toxicity when treated with standard doses of 6MP (50–75 mg/m2/day). The frequency of TPMT alleles has not previously been evaluated in the Russian Federation. We determined the allele frequency and the pattern of mutant TPMT alleles in Russian children with hematologic malignancies (leukemia/lymphoma) and other diseases, and in healthy adults, in different areas of the Russian Federation. A microchip was designed to detect the main mutant alleles of human TPMT (TPMT*2, *3A, *3B, *3D, *7, and *8) and was used to assay 982 samples. Only 3 inactivating mutations of the TPMT gene were identified: 45 subjects (4.6%) had TPMT genotype *1/*3A, 8 (0.8%) had *1/*3C, and 2 (0.2%) had *1/*2. No homozygous genotypes and no mutant *3B, *3D, *7, or *8 alleles were found. The estimated frequency of the wild-type allele was 97.1% (95% CI, 93.8%–100%) and that of variant alleles was 2.9% (95% CI, 0%–6%). Most individuals with the *1/*3A genotype were of Slavic background, and genotype *1/*3C was found in a buryat (Mongolian race) and in residents of the North Caucasian and the Urals regions. These findings indicate ethnic differences in the frequency of nonfunctional TPMT alleles. The overall frequency of mutant TPMT genotypes in the Russian Federation is 5.6%, which is similar to that observed in the white populations of the United States and Europe.

Description: Conventional therapies do not provide satisfactory control of multiple sclerosis (MS) due to their inability to eradicate self-specific T cell clones. During the last decade high-dose immunosuppressive therapy (HDIT) with autologous haematopoietic stem cell transplantation (AHSCT) has been used with increasing frequency as a therapeutic option for MS patients. We have identified 4 strategies of HDIT+AHSCT depending on the stage in disease process: early, conventional, late, and salvage. In the current study we aimed to study clinical and patient-reported outcomes in MS patients after different strategies of HDIT+AHSCT. One hundred and twenty patients with MS (secondary progressive – 53 patients, primary progressive –21, progressive-relapsing – 9, relapsing-remitting – 37) were included in this study (mean age - 33.0, range: 17–54; male/female – 53/67). Thirty seven patients underwent early transplantation (EDSS 1.0–3.0), 72 patients – conventional transplantation (EDSS 3.5–6.0), 6 patients – late transplantation (EDSS 6.5–7.0), and 5 patients – salvage transplantation (EDSS 7.5–8.5). Median EDSS at baseline was 5.0 (range 1.5 – 8.0). The mean follow-up duration was 18 months (range 6 – 108 months). Neurological and quality of life (QoL) evaluation was performed at baseline, at discharge, at 3, 6, 9, 12 months, and every 6 months thereafter following AHSCT; MRI examinations - at baseline, at 6, 12 months, and at the end of follow-up. Notably, transplantation procedure was well tolerated by the patients with no transplant-related deaths. The efficacy analysis was performed in 79 patients. At 6 months post transplant the following distribution of patients according to clinical response was observed: 42 patients (53%) achieved an objective improvement of neurological symptoms; 37 patients (47%) had disease stabilization. At long-term follow-up clinical response was classified as improvement in 40 patients (50.6%); stabilization in 34 patients (43.1%); progression in 5 patients (1 patient after early transplantation, 3 patients after conventional transplantation, and 1 patient after late transplantation). No active, new or enlarging lesions were registered in patients without disease progression. Furthermore, out of 44 patients included in QoL analysis 39 exhibited improved QoL 6 months post-transplantation. Further QoL improvement was observed at longer follow-up in patients without disease progression. All the patients without disease progression were off therapy throughout the post-transplant period. Thus, AHSCT appears to be an effective treatment for MS both in terms of clinical and patient-reported outcomes. Further studies should be done to establish the best timing for transplantation and to validate high-dose immunosuppressive therapy with ASCT regimens in patients receiving early, conventional, late and salvage transplantation.

Description: Abstract 3518 Poster Board III-455 Introduction Thrombocytopenia [TCP] is a complication of advanced chronic liver disease [CLD]. We examined clinical factors related to the development of severe TCP among patients with initially normal platelet counts using laboratory, clinical, and administrative data from the Veterans Administration Healthcare System, New England region. Methods All patients with a diagnosis of CLD or hepatitis C who were seen in a VA hospital or in a VA outpatient clinic between 10/2002 and 9/2008 were included in this analysis. CLD diagnosis definitions included: 1) ICD9-CM code of 51.0 – 571.9, excluding 571.1at a clinic visit or hospital discharge 2) positive hepatitis C [Hep-C] antibody, and 3) elevated ALT (〉 84 IU/dL) on two occasions at least 60 days. Subjects were required to have normal platelet counts (platelet count 〉150,000/μL) at baseline. Subjects were excluded if they had the following: 1) hepatorenal syndrome, coagulation disorders, immune TCP purpura, lupus erythematosis, rheumatoid arthritis, pernicious anemia, aplastic anemia or septicemia. Severe TCP was defined as a platelet count 〈 50,000/μL. Risk factors evaluated for the development of TCP included age, comorbidities (diabetes, alcoholic and/or drug dependence, chronic kidney disease, Hepatitis-C [Hep-C] status, hepatorenal syndrome, congestive heart failure [CHF], coronary artery disease, hypertension), resource utilization (infectious disease, gastrointestinal [GI] nephrology and/or hematology clinic visit), and current laboratory values (hemoglobin, ALT and MELD Score). All potential predictors were treated as time-varying exposures over the follow-up and updated at each interval. Relative risk was estimated using logistic regression with pooled repeated observations, using 3-month periods where risk factors at the beginning of a 3-month interval were used to predict severe TCP in that interval. Results 6,102 subjects with Hep-C contributed a median follow-up of 1,414 days and had 82 events (1.34%) during the study. 5,358 Non Hep-C subjects contributed a median follow-up of 1,238 days and had 97 events (1.81%) during the study. The median platelet count among CLD patients at baseline was 238,000/μL (IQR 203,000/μL- 282,000/μL) and among Hep-C patients at baseline was 240,000 (IQR 205,000/μL- 285,000/μL). After multivariate adjustment, alcohol dependence [Odds ratio = 2.24, 95% confidence interval: (1.14, 4.39)], CHF [2.6 (1.05,6.43)], 5 unit change in MELD [1.13 (1.05,1.21)], GI clinic visits [1.89 (0.93,3.87)] and anti retroviral therapy [3.84 (1.60, 7.61)] increased the odds of severe TCP; while drug dependence [0.62 (0.34, 1.13)] and increasing hemoglobin [0.74 (0.65,0.84)] reduced the odds of severe TCP. In non Hep-C subjects, alcohol dependence [2.88 (1.58,5.25)], CHF [1.81 (0.9, 3.65)], 5 unit change in MELD [1.12 (1.03, 1.23)], hematology clinic [3.64 (1.91, 6.93)] current anti-retroviral therapy [6.65 (1.96, 22.6)] and ALT 42-84g/dL vs. ALT 〈 42g/dL [1.55 (0.78, 3.06)] increased the odds of severe TCP; while drug dependence [0.46 (0.17, 1.22)], increasing hemoglobin [0.68 (0.60, 0.78)] and ALT 〉 84g/dL vs. ALT 〈 42 reduced the odds of severe TCP. Conclusions In a population of veterans with chronic liver disease or hepatitis C infection with normal platelet counts 〉 150,000/μL at baseline, we estimate the incidence of development of severe thrombocytopenia to be between 1 and 2% over an average of 3 years of follow-up. Key clinical predictors of development of severe TCP included prevalent CHF diagnosis and changing MELD score. Specialist care referrals to hematologists and gastroenterologists, also identified patients likely to become severe TCP cases. As expected, antiretroviral therapies were associated with an increased risk of severe TCP over the follow-up. Acknowledgments GlaxoSmithKline provided research support for this project, with no restrictions on publication. The study was conducted using resources of the VA Cooperative Studies Program and VA Boston Healthcare System. Disclosures: Lawler: GlaxoSmithKline: Research Funding. Horblyuk:GlaxoSmithKline: Employment. Grotzinger:GlaxoSmithKline: Employment, Research Funding.

Description: Ninety ET patients diagnosed according to the PVSG criteria were enrolled in a phase II study (sponsored by the Schering-Plough Company) designed to evaluate the efficacy, safety and tolerability of a two years treatment with PEG Interferon alpha-2 b (PEG Intron). The patients, observed in 16 Italian Centres belonging to the GIMEMA Cooperative Group and judged at high risk, had been previously treated with cytoreductive (97%) and antiplatelet (91%) drugs. At the study start the patients, 60 F and 30 M, mean age 45 years, showed splenomegaly in 22% of cases. The Hematological Response (HR: PLT