ALBERT

Description: Background T-cell large granular lymphocytic leukemia (T-LGLL) is a rare lymphoproliferative disorder of cytotoxic T cells (CTLs) and often associated with autoimmune disorders. The signal transducer and activator of transcription 3 (STAT3) is an oncogene, and its activation plays a key role in cell signaling transduction pathways in many types of cancer. The aim of the current study was to analyze the characteristics of T-LGLL. Methods We did this by determining the mutation status of STAT3 in 28 patients presenting with T-LGLL and evaluating STAT3 status in association with serum level of lacticdehydrogenase (LDH) and β2-microglobulin (β2-MG). Flow cytometric analysis for immunophenotype and TCR variable β-chain (Vβ) was performed in the patients. Results FC-Vβ analysis was performed in 26 patients, and 22 (84.6%) patients had a restricted Vβ reactivity pattern, with predominance of a single Vβ mAb reactivity. There was no significant difference between serum LDH levels, gender, age or symptoms at diagnosis (P=0.062), lymphocytosis, anemia (P=0.057), thrombocytopenia, splenomegaly, LGL count or STAT3 mutation status. However, high β2-MG levels (P=0.005), neutropenia (P=0.018) and pure red blood cell aplasia (PRCA) (P=0.001) all displayed a significant association with STAT3 mutations. In univariate analysis, treatment-free survival (TFS) was affected by STAT3 mutation status (P=0.008) and β2-MG (P=0.006). In multivariate analysis, only anemia (P

Description: Abstract 4387 Chronic lymphocytic leukemia (CLL) is one of the most common lymphoid malignancies in the Western countries, however, infrequent in Asian populations. Although the median survival is around 10 years, CLL is a disease with an extremely variable clinical course with overall survival times ranging from months to decades; some patients never need treatment, while others require intensive treatment early after diagnosis. Some factors, such as clinical stages, IGHV mutational status, cytogenetic abnormalities, ZAP-70, and the expression of CD38 in leukaemic cells, were strong indicator of prognosis in CLL. However, the prognostic factors of Chinese patients with CLL compared with the Western countries have not yet been clarified. The aim of this study was to explore the influence of factors on the prognosis of Chinese patients with CLL. One hundred and twenty-nine patients with CLL were enrolled in this study. Multiplex PCR and sequencing, fluorescence in situ hybridization (FISH), and flow cytometry were used to detect IGHV mutational status, cytogenetic abnormalities, and the expression of ZAP-70 and CD38, respectively. A panel of FISH probes included 13q14 (D13S319), 17p13 (p53 gene), 11q23 (ATM gene), 6q23(MYB gene), the centromere of chromosome 12 (D12Z3) and 14q32 (IGHC/IGHV). In 129 CLL patients, according to the Binet clinical staging system, 65 (50.4 %) patients were in Binet A, 28 (21.7 %) in Binet B and 36 (27.9 %) in Binet C. Eighty-four (65.1%) patients had mutated IGHV, and 45 (34.9%) had unmutated IGHV. The most frequently expressed VH gene family was found to be VH3 (50.4%) followed by VH4 (32.6%), VH1 (10.9%), VH2 (2.3%), VH5(2.3%) and VH7 (1.6%), with no expression of VH6 gene families. VH1-69 and VH3-21 which commonly overused in Western CLL patients were very low in our cohort (0.8% and 3.1%, respectively). Molecular cytogenetic aberrations were found in 94 patients (72.9%) and 36 patients (27.9%) with more than two abnormalities. The most frequent abnormalities detected in our patients was del(13q14), with an incidence of 53.0%, followed by 14q32 translocation of 20.2%, +12 of 18.3%, del(11q23) of 10.8%, del(17p13) of 10.o%, and del(6q23) of 6.1%. Forty-one patients (31.8%) were positive for ZAP-70 (≥20%), and 51 patients (39.5%) were positive for CD38 (≥30%). With a median follow-up of 32 months (range, 4-58 months), eight patients (6.2%) died (CLL-related deaths). In univariate analysis for survival, advanced Binet stage (P=0.023), unmutated IGHV status (P=0.002), deletions of 17p13 or 11q23 (P=0.003), high expression of ZAP-70 (P=0.034), and high expression of CD38 (P=0.046) were poor prognostic factors. The prognostic factors with statistical significance were further used in a two-variables Cox analysis, which comparing unmutated IGHV status to other prognostic factors individually to show prognostic independence. The unmutated IGHV status were the independent prognostic factors and strongly associated with OS. This study demonstrates that the frequencies of IGHV gene families indicated significant difference in Chinese CLL patients compared with Western patients, suggesting involvement of ethnic and/or environmental factors in CLL disease initiation. The unmutated IGHV status, Binet clinical stages, Chromosomal aberrations of del(17p13) and del(11q23), high expression of ZAP-70 and CD38 have been shown highly predictive prognostic value for Chinese patients with CLL. Disclosures: No relevant conflicts of interest to declare.

Description: Abstract 3450 Poster Board III-338 Chronic lymphocytic leukaemia (CLL) is characterized by a progressive accumulation of monoclonal B lymphocytes in the bone marrow, lymphatic organs and blood. Even with the use of newer therapies, many patients with CLL develop progressive, treatment-resistant disease. Rituximab (RTX), a monoclonal antibody, targets CD20 on B lymphocytes and widely used in indolent B cell neoplasms. The lower density of CD20 on CLL cells may play a role, however, the causes of the lesser susceptibility of CLL to RTX remain poorly understood. Complement deficiencies have been identified in many CLL patients and appear to be more pronounced in patients with more advanced disease. Since complement mediated cell lysis is one of the major mechanisms of RTX action, we hypothesized that the therapeutic effect of RTX in CLL may be enhanced by the provision of complement through concurrent administration of fresh frozen plasma (FFP). Between August 2008 and April 2009, twelve patients with refractory advanced CLL were enrolled in this open clinical trial. All had been previously treated with fludarabine and two also failed treatment with RTX. The median age patients was 57 years (range, 48-82 years). According to the Binet staging system, 2 patients were in Binet B, 10 patients in Binet C. Florescence in situ hybridization (FISH) studies identified del(13q14) in 8 patients, and 4 patients with del(13q14) as the sole abnormality. Trisomy 12 was found in 3 patients, del(11q22.3) in 2 paitents, del(17p13) in 2 patients, del(6q23) in 1 patient, and IgH translocation in 1 patient, respectively. By flow cytometry, CD38 was positive in 9 patients, and ZAP-70 positive in 8 patients. Immunoglobulin heavy chain variable gene (IgVH) mutational status was detected by multiplex PCR, and six patients had unmutated VH. Two units of FFP followed with RTX 375 mg/m2 as a single agent, repeated every 1-2 weeks up to a total of 2-5 cycles. Complete response (CR) was achieved in 4 patients (33.3%), partial remission (PR) in 6 patients (50.0%). A major improvement of clinical signs and symptoms was achieved in all patients. Lymphocyte counts dropped markedly followed by shrinkage of lymph nodes and spleen and improvement of the anaemia and thrombocytopenia. This could be maintained over 9 months (range, 3-11 months) without progression. Toxicity was minimal and the treatment was well tolerated in all cases. It was show that RTX combining with FFP may provide a useful therapeutic option in patients with refractory advanced CLL. Disclosures No relevant conflicts of interest to declare.

Description: Objective To investigate serum thymidine kinase (TK) level in Chineses patients with chronic lymphocytic leukemia (CLL) and its correlation with other prognostic factors, including Binet stages, absolute lymphocyte count (ALC), lactate dehydrogenase (LDH), immunoglobulin heavy-chain variable region (IgVH) gene mutation status, ZAP-70 protein and CD38 expression level, and cytogenetic aberrations. Methods Serum TK1 level in 39 CLL patients was detected by TK monoclonal antibody (Anti-TK mAb) and enhanced chemiluminecence (ECL). IgVH mutation status was detected by multiplex PCR and sequencing of purified PCR amplification products. A panel of monoclonal antibodies and multiparametic flow cytometry were employed to immunophenotype and determine the expression of ZAP-70 protein and CD38. A panel of probes and interphase fluorescence in stu hybridization (FISH) were used to detect cytogenetic aberrations including 6q-, 11q-, +12, 13q-,17p- and IgH translocation. Results The level of TK1 was higher in CLL patients that in normal control (P

Description: Chronic lymphocytic leukemia (CLL) is the most common type of adult leukemias in the Western countries, however, infrequent in the Eastern. Autoimmune hemolytic anemia (AHA) is a complication in chronic lymphocytic leukemia (CLL). The direct antiglobulin test (DAT) may be positive at some time during the disease course in up to 35% of cases, but overt AHA occurs less frequently. The aim of the study was to explore the prognostic impact of positive DAT in Chinese patients with CLL and its correlation with other prognostic factors, including Binet stages, lymphocyte count in peripheral blood, lactate dehydrogenase (LDH), β2-microglobulin (β2-MG), IgVH mutation status, ZAP-70, CD38 and cytogenetic abnormalities. Out of the 80 Chinese patients with CLL, positive DAT was found in 21 (30.6%) cases. The incidence of positive was 12.5% in Binet A, 23.8% and 44.4% in Binet B and C, respectively. The incidence of positive DAT was significantly increased at Binet C, compared with Binet A (P=0.006), and the presence of higher LDH and β2-MG levels correlated strongly with positive DAT (P=0.006 and P=0.004, respectively). Patients with unmutated IgVH genes had higher incidence of positive DAT than did patients with IgVH mutations (P=0.042), and positive DAT was also associated with higher level of ZAP-70 and CD38 (P=0.004 and P