ALBERT

Description: In respect to B lymphocyte-mediated immunity, characteristics of human cord blood are low counts of mature B lymphocytes, deficient expression of CD40L and cytokine production in CD4+ T lymphocytes, defect in the isotype switch of immunoglobulin and the activation of B lymphocytes, and low IgG production of B lymphocytes. These characteristics of the B lymphocyte from human cord blood lead to a delayed B lymphocyte-mediated immune reconstitution and an increased susceptibility to infections after a cord blood transplantation. The mechanism of immunological recostitution after cord blood transplantation has been examined from a variety of viewpoints in experimental models as well as clinical studies. However, problems of sustained immunodeficiency after cord blood transplantation remain to be resolved. The aim of the present study is to establish culture conditions that support the effective B lymphocyte expansion of human cord blood using IL-4, IL-10, and CD40L, to which cytokines are defected in B lymphocyte of human cord blood, and established conditions are compared to previously established cytokine combinations, TPO+SCF+FL in our Lab (Br J Haematol 107:176–185, 1999 & Stem Cells 21:228–235, 2003). To elucidate the effective B lymphocyte-mediated immune reconstitution of cord blood after ex vivo expansion, mononuclear cells, separated from density gradient of Ficoll system, and CD34+ purified cells, isolated from immunomicrobead(MiniMACS) system, were cultured with various combinations of cytokines (TPO+FL+SCF and/or IL-4, IL-10 and CD40L) for 2 weeks or 4 weeks. This then allowed for cytometric analysis after immunofluorescence stain with CD34, CD38 (for HSC analysis) and CD19, IgG and IgM (for B lymphocyte-mediated immune reconstitution) and CD4 (for T helper cell) and CD25 (for lymphocyte activation assay) to be performed. In the B lymphocyte expansion aspect, the immunoglobulin expression, and functional activity, expansion with the TPO+FL+SCF+IL-4+IL-10 combination showed best results in the expression of CD19, CD25, IgG, and IgM. However, the addition of CD40L to those culture condition did not increase expression of CD19, CD25, IgG, and IgM after the expansion of human cord blood. Expansion of CD34+ purified cells was superior to MNCs in the expression of CD19, CD25, IgG, and IgM. In consideration for the duration of cultures, the 2 week culture was superior to the 4 week culture with respect to graft stemness (CD34+CD38- fraction). Our data suggests most superior results were observed from the ex vivo expansion of CD34+ purified cells cultured for 2 weeks with TPO+FL+SCF+IL-4+IL-10, in the B lymphocyte-mediated immune reconstitution and graft stemness aspect. The results of this study warrant further investigation on effective B lymphocyte-mediated immune reconstitution after cord blood transplantation in vivo using ex vivo expanded cord blood.

Description: The aim of this retrospective cohort study was to analyze the impact of surgery on the outcomes and qualities of life (QOL) in patients with intestinal diffuse large B-cell lymphoma (DLBCL). We assessed 345 patients with either localized or disseminated intestinal DLBCL and compared them according to treatment: surgical resection followed by chemotherapy versus chemotherapy alone. In patients with localized disease (Lugano stage I/II), surgery plus chemotherapy yielded a lower relapse rate (15.3%) than did chemotherapy alone (36.8%, P 〈 .001). The 3-year overall survival rate was 91% in the surgery plus chemotherapy group and 62% in the chemotherapy-alone group (P 〈 .001). The predominant pattern in the chemotherapy group was local relapse (27.6%). When rituximab was used with cyclophosphamide, doxorubicin, vincristine, and prednisolone (CHOP), there was no improvement of the outcomes in patients treated with primary surgical resection. The QOL of patients who underwent surgery and chemotherapy was lower than chemotherapy alone, but its difference was acceptable. Multivariate analysis showed that surgical resection plus chemotherapy was an independent prognostic factor for overall survival. Surgical resection followed by chemotherapy might be an effective treatment strategy with acceptable QOL deterioration for localized intestinal DLBCL. This study was registered at www.clinicaltrials.gov as #NCT01043302.

Description: There is abundant evidence that S1P can function as a second messenger important for regulation of calcium homeostasis, cell growth, and suppression of apoptosis. In many cases, the intracellular level of S1P and ceramide, another important sphingolipid metabolite associated with cell death and cell growth arrest, coordinately determine cell fate. N, N-dimethylsphingosine(DMS) potentiates TNF- and FasL- induced apoptosis in the human acute leukemia jurkat, U937 and HL-60 cell line, which suggests that combining DMS with cytotoxic drugs might be a useful chemotherapeutic approach. In this study, we investigated the role of S1P on proliferation, survival and its signaling mechanisms on the NB4 cell, the human acute promyelocytic leukemia cell line, which has chromosomal abnormality, t(15:17). NB4 cells were exposed to S1P at various concentrations (1, 5, 10 μM) for 48 hours, and then the cell growth was determined by MTT assay. We studied whether NB4 cells have S1P receptors or not. To investigate the signaling mechanisms of S1P on the NB4 cell, we used MAP kinase inhibitors and a PI3K inhibitor including PD98059, U0126 (specific ERK pathway inhibitor), SB203580 (specific p38 pathway inhibitor), and LY294002 (specific PI3K pathway inhibitor). We also examined the effect of pertussis toxin (PTX), a Gi protein inhibitor, on the proliferation of NB4 cells after S1P treatment. RT-PCR analysis revealed a putative S1P receptor, Edg-3 mRNA expressed in NB4 cells. In the MTT assay, S1P inhibited cell proliferation in a dose-dependent manner. At 10 μM, NB4 cell proliferation was most inhibited. In addition, we found that the inhibitory effect of S1P on the NB4 cell proliferation was inhibited by PD98059, U0126, and PTX. However, SB203580 and LY294002 had not caused an inhibitory effect of S1P on the NB4 cell proliferation. In contrast to previous knowledge that S1P increases the survival of leukemic cells because apoptosis is prevented by caspase inactivation, our data suggests that S1P inhibits NB4 cell proliferation, through ERK pathway activation, not p38 nor JNK pathways. In addition, the pertussis toxin(PTX)-sensitive GTP-binding protein-coupled receptor, Edg-3, seems to be involved in the antiproliferative signaling of S1P to the NB4 cell. Understanding the signaling mechanisms of anti-proliferation by S1P can uncover new therapeutic targets for APL and the results of this study warrant further investigation on synergism of S1P with known therapeutic agent, ATRA and Arsenic trioxide on APL cells.

Description: Background: Due to the rarity of the disease, prospective clinical trials on optimal treatment of intestinal non-Hodgkin’s lymphoma have been scarcely performed. Methods: Patients with; age 〉 18 years, pathologically proven diffuse large B cell lymphoma (DLBL), normal bone marrow, liver, renal and cardiac function were eligible. Patients with Burkitt’s lymphoma, lymphoblastic lymphoma, mantle-cell lymphoma, or infection with human immunodeficiency virus were excluded. All patients were staged after surgery according to the Ann Arbor classification modified by Musshoff et al. and Paris staging system (TNM system). Postoperative chemotherapy with CHOP regimen was administered for 6 cycles. CHOP consisted of cyclophosphamide 750 mg/m2 d1, doxorubicin 50 mg/m2 d1, vincristine 1.4 mg/m2 (max 2.0 mg/m2) d1, and prednisolone 100mg d1–5 repeated every 3 weeks. Results: From 1999 to 2004, 41 patients were enrolled. The median age was 50 years (range, 26 – 84) and male:female ratio was 29:12. All patients had clinical stage I/IIE DLBL and underwent surgical resection. Of the 41 patients, 3 patients refused postoperative CHOP chemotherapy. The 5-year DFS and OS were 83.3% and 89.1%, respectively. Surgicopathologic staging of all patients according to the Ann Arbor classification revealed 34 patients with stage I, 11 patients stage II1, 1 patent stage II2 and 3 patients with stage IIE. The Ann Arbor stage significantly predicted the survival of intestinal DLBL (p = 〈 0.0001). The T staging according to Paris staging system did not correlate with survival (p = 0.1456) while the N staging showed significant predictability (p =

Description: Abstract 4992 Introduction Intestine is the one of commonly involved extranodal sites of non-Hodgkin's lymphoma (NHL). Thus, small and large Intestine account for approximately 30-40% of primary gastrointestinal tract lymphoma. More than 70% of intestinal lymphoma presents as localized disease, and surgery such as bowel resection is performed in many patients with intestinal lymphoma for diagnosis and treatment. However, it is still unclear whether surgical resection followed by chemotherapy is superior to systemic chemotherapy alone in terms of treatment outcome and quality of life (QOL). Thus, we retrospectively analyzed the clinical features and treatment outcome of patients with NHL of intestine, and performed a multicenter cross-sectional study about the QOL in survivors of intestine NHL. Patients and methods We evaluated 463 patients with intestine NHL from 15 hospitals affiliated with the Consortium for Improving Survival of Lymphoma (CISL) in Korea. The QOL was assessed in 84 survivors who completed their treatment using the EORTC QLQ-C30 questionnaire. Results The median age was 55 years old (range 15-92), and male to female ratio was 1.79:1. 389 patients (84.0%) had the ECOG performance status less than 2. More than a half of patients (59.8%) presented as a localized disease: Ann Arbor stage IE (n = 127, 27.7%) and IIE (n = 150, 31.3%). The cases involving two or more than two extranodal sites were found in 26.6%, and the elevation of serum LDH was observed in 173 patients at diagnosis (37.4%). Thus, the majority of patients had low risk of IPI (50.8%, n=235) while high risk was 10.2% (n=47). The presence of B symptoms and invasion of bone marrow were relatively less frequent event (19.0% and 9.7%, respectively). The most common histological subtype was DLBCL (71.3%, n=330), and the frequency of other subtypes was as follows: MALT lymphoma (8.2%, 38), Burkitt lymphoma (6.7%, 31), PTCL (6.7%, 31), mantle cell lymphoma (3.7%, 17), and others. The frequency of involved sites was as follows: small intestine including terminal ileum and jejunum (61.7%), large intestine (26.6%), and small and large intestine (11.7%). The major treatment modality was surgery followed by chemotherapy or chemotherapy alone. Thus, 205 patients received surgery followed by chemotherapy while 170 patients received chemotherapy alone. The 5-year overall survival (OS) was 69.8% (95% CI: 64.18-72.82), and 5-year progression-free survival (PFS) was 67.2% (95% CI: 61.72-70.53). Because DLBCL accounts for more than 90% of patients treated with surgery plus chemotherapy or chemotherapy alone, the survival outcome was compared in stage IE/IIE of DLBCL as a subgroup analysis. 5-year OS was significantly higher in the group with surgery plus chemotherapy (86.67%, 95% CI: 81.43-91.9) than the group with chemotherapy alone (66.23%, 95% CI: 56.96-75.49, P 〈 0.001). However, there was no significant difference of OS in DLBCL patients with stage III/IV. When we compared the survival outcome according to the use of rituximab-CHOP versus CHOP, the addition of rituximab failed to show additional survival benefit in DLBCL patients with stage IE/IIE. The QOL of survivors was not significantly different based on the treatment modality except a tendency of better physical and role functioning in patients treated with chemotherapy alone. The global health status was comparable between two groups. Conclusion Surgery followed by chemotherapy might be a better treatment strategy for localized non-Hodgkin lymphoma of intestine in terms of survival and quality of life. Disclosures No relevant conflicts of interest to declare.

Description: Purpose: The International Prognostic Index (IPI) and Follicular Lymphoma Prognostic Index (FLIPI) are used as prognostic indices for NHL and indolent lymphoma. However, marginal zone B-cell Lymphoma (MZL) evidences a distinctive clinical presentation and a natural course; thus, in this study, we attempted to devise an adequate prognostic index for MZL. Patients and method: From 1990 to 2005, 205 patients diagnosed with MZL were retrospectively reviewed. After analysis of the prognostic factors, progression free survival (PFS) and overall survival (OS), we constructed a prognostic index of MZL (MZLPI) via the summation of each factor. We then compared PFS and OS with IPI, FLIPI, and MZLPI. Results: According to our multivariate analysis of PFS and OS of MZL, nodal MZL, ECOG performance ≥ 2 and advanced stage were composed of MZLPI. MZLPI was grouped as follows: score 0 as a low risk group, score 1 as an intermediate risk group, and score ≥2 as a high risk group. The PFS curve, according to MZLPI results, evidenced a more discriminated pattern than IPI and FLIPI, and this was especially true in the intermediate risk group. In OS, MZLPI (P=0.0007) evidenced a more discriminated pattern than IPI (P=NS) or FLIPI (P=0.0044). Conclusion: MZLPI, which is constructed of relatively simple factors, may represent a useful prognostic index for the prediction of PFS and OS in MZL, and may also be used as a substitute for IPI or FLIPI.

Description: Purpose: Nongastric marginal zone B-cell lymphoma (NG-MZL) is a relatively uncommon indolent lymphoma. We report a retrospective analysis of 247 patients with .NG-MZL, presenting their clinical features and therapeutic outcomes. Methods: From 1990 to 2005, a total of 247 patients with histologically confirmed NG-MZL were analyzed. Results: The median age was 49 years (range, 13–89 years). The study involved 129 males (52.2%) and 118 females (47.8%) The most common involving site was orbit and ocular adnexa (48.6%) followed by lymph node and lymphatic organs (17.8%), bowel (9.3%), lung (6.1%), thyroid gland (4.9%), salivary gland (4.5%) in the decreasing order of frequency. Ann Abor stage I/II disease was present in 78%(167 out of 215). BM involvement was less than 10%(19 out of 211). B symptom was observed in only 2%. One and eighty-six patients out of 208 were in low or low-intermediate risk group (89%) according to international prognostic index(IPI). Eighty percents (172/215) were in low risk group in follicular lymphoma international prognostic index (FLIPI). Patients were treated with a variety of therapeutic strategies. Complete and partial remissions were achieved in 139(92%) and 8(5.3%) of the 151 stage I/II patients, respectively, with an overall response rate of 97.3%. Especially, radiation containing treatment achieved 96% CR rate (108 out of 113). In 38 patients with stage III/IV, CR and PR were achieved in 17(44.7%) and 11(28.9%). The estimated 5-year overall survival (OS) and progression free survival (PFS) were 93.8% and 70.1%, respectively. Although anthracyclin contaning regimen could achieve higher CR rate, it did not improve PFS. Stage III/IV, low hemoglobin, high/high-intermediate IPI, poor risk FLIPI and nodal MZL were poor prognostic factors for PFS in multivariate analysis. Conclusion: NG-MZL is an indolent disease. For localized disease radiation achieved excellent local control. Anthracyclin containing chemotherapy cannot improve outcome. Both IPI and FLIPI can be applied to predict the prognosis.

Description: Background: Hepatic veno-occlusive disease (VOD) is a devastating regimen-related toxicity after hematopoietic stem cell transplantation (HSCT). There is no safe and proven therapy for established VOD, and focus has been on its prevention. Previous studies have shown that ATIII level is markedly decreased in patients with VOD. We conducted a prospective, randomized trial to compare heparin plus ATIII versus heparin alone in HSCT recipients. Methods: A clinical diagnosis of hepatic VOD was defined as satisfying two of the following criteria: bilirubin 〉 2.0 mg/dl, unexplained weight gain of 〉 2% from baseline, and hepatomegaly. The levels of PAI-1, TFG-β and TNF-α were measured at weekly basis for correlative analysis. Patients were randomized to receive or not receive prophylactic ATIII 1000U twice daily from day 1 until day +14 post-HSCT plus heparin 5U/kg/hr from day 1 until day +21 or discharge after HSCT. Results: Thirty patients were enrolled for the study. Fourteen patients received heparin + ATIII and 16 patients received heparin alone for VOD prophylaxis during HSCT. There were 9 AML, 6 NHL, 4 ALL, 3 CML, 2 ABL, 2 MM, 1 MDS, 1 HD, 1 AA and 1 SLL patient. Of the 30 patients enrolled, 13 patients received sibling allogeneic HSCT, 13 autologous, 3 unrelated allogeneic and 1 cord blood HSCT. VOD occurred in 4 patients (25%) in the heparin alone group and none in the heparin + ATIII group (0%, P = 0.044). The incidence of bleeding complication was higher in the heparin alone group (4/16, 25%) than the heparin + ATIII group (1/14, 7%) without statistical significance (P = 0.190). The mean baseline level of PAI-1 was slightly higher in the heparin + ATIII group (30.9 ± 31.0 ng/ml) when compared to the heparin group (18.9 ± 13.0 ng/ml, P = 0.062). The PAI-1 level decreased markedly in the heparin + ATIII group (23.0 ±10.1 ng/ml) by the third week following VOD prophylaxis therapy (heparin group; PAI level 40.3 ± 40 ng/ml, P = 0.010). The mean baseline levels of TGF-β and TNF-α did not differ significantly between the two groups. There was no VOD-related death. Conclusions: Although it was a preliminary analysis, the combination of ATIII and heparin for VOD prophylaxis in HSCT recipients seems to be more effective than heparin alone without increasing the hemorrhagic complication.