Type Of B-cell non-Hodgkin Lymphoma Research Articles

Quantitation of the JAK2 V617F Mutation in Microdissected Bone Marrow Trephines: Equal Mutational Load in Myeloid Lineages and Rare Involvement of Lymphoid Cells

The JAK2V617F mutation is an essential oncogenic event in Philadelphia negative chronic myeloproliferative disorders (Ph-cMPD). It is still unclear how a unique tyrosine kinase mutation can give rise to the broad clinical and morphologic spectrum of Ph-cMPD. One possible explanation could be differences in the JAK2V617F gene dosage, or different maturation stages on which myeloid lineages are affected by the mutation. The extent of lymphoid lineage involvement in JAK2V617F-positive cMPD is still controversial. We comparatively studied the zygosity status of microdissected megakaryocytes, nonmegakaryocytic hematopoietic cells, and reactive as well as neoplastic lymphoid nodules from bone marrow trephines of 61 patients with Ph-cMPD. The presence of the mutation and mutant gene dosage were determined by allele-specific polymerase chain reaction and TaqMan analysis, respectively. The mutation was detected in 22/32 (68%) cases of essential thrombocythemia, all cases of polycythemia vera, and 4/8 (50%) idiopathic myelofibrosis. Comparison of whole bone marrow sections and the different myeloid lineages showed similar percentages of the mutated allele. Restriction to a particular lineage or major differences in allele dosage were not observed, except for 2 cases in which megakaryocytes revealed a higher frequency of the mutated allele. A heterozygous JAK2V617F mutation was detected in 3/8 "reactive" lymphoid nodule in patients with Ph-cMPD, whereas all concomitant non-Hodgkin lymphoma of B-cell type were negative. These results demonstrate that different myeloid lineages usually show similar frequencies of the JAK2V617F allele. The occasional detection of JAK2V617F in benign lymphocytes points to involvement of the lympho-myeloid stem cell.

Non-Hodgkin lymphoma as a cause of obstructive jaundice with simultaneous extrahepatic portal vein obstruction: A case report

Non-Hodgkin lymphoma is a rare cause of biliary obstruction. To the best of our knowledge, non-Hodgkin lymphoma in the peripancreatic region causing obstructive jaundice with simultaneous portal vein (PV) invasion has not yet been reported. We present a 50-year-old patient with obstructive jaundice whose extrahepatic portal vein was obstructed by the invasion of a peripancreatic non-Hodgkin lymphoma. The patient denied any other symptoms such as recurrent fever, night sweat and loss of body weight. Computed tomography (CT) revealed a 10 cm mass in the retroperitoneal space behind the head of the pancreas causing obstruction of the distal bile duct and the PV. A pylorus-preserving pancreaticoduodenectomy combined with a PV resection was performed. The PV was reconstructed using an autologous right internal jugular vein graft. The resected specimen showed endoluminal invasion of both the bile duct and the PV. Histological examination showed the mass consisting of diffuse sheets of large malignant lymphoid cells. These cells were positive for CD20 and CD79a, partially positive for CD10, and negative for CD3, CD4, CD5, CD8 and CD30. The pathologic diagnosis was diffuse large B-cell type non-Hodgkin lymphoma and the patient was transferred to the Department of Hematology and Oncology for chemotherapy. He received four cycles of combined chemotherapy including cyclophosphamide, doxorubicin, vincristine and prednisone plus rituximab, and three cycles of intrathecal chemoprophylaxis including methotorexate, cytosine arbinoside and prednisone. The patient is alive with no evidence of the disease for 7 mo after operation and will receive additional courses of chemotherapy.

Gene Expression Analysis of B-Cell Post Transplant Lymphoproliferative Disorders Provides Insights into Disease Biology.

Background: B-cell post transplant lymphoproliferative disorders (PTLDs) encompass a diverse group of entities, the majority representing EBV-driven lymphoproliferations, which arise in the setting of iatrogenic immunosuppression. Their pathogenesis, especially the genetic relationship of PTLD with other types of B-cell non-Hodgkin lymphomas (B-NHLs), is not well understood and the association of polymorphic PTLD (P-PTLD) with monomorphic PTLD (M-PTLD) is presently unclear. Thus, in order to address these issues we analyzed gene expression profiles of PTLD and compared them with those derived from other types of B-NHL, normal B-cell subsets, and B-cell derived cell lines.Material and methods: Paraffin embedded and H&E stained sections were used for morphologic analysis and PTLD were classified according to the WHO classification. Phenotypic characterization was performed using a comprehensive panel of antibodies and in situ hybridization for EBER. Frozen tissue from PTLD, different B-NHL, purified normal B-cell subsets, EBV-transformed lymphoblastoid cell lines (LCLs) and multiple myeloma cell lines, was used to generate complementary RNA (cRNA) that was hybridized to HGU95 arrays (Affymetrix). Gene expression data were analyzed using unsupervised and supervised clustering algorithms and classifiers were generated to determine the relationship between PTLD and cells corresponding to different stages of mature B-cell development.Results: The 12 PTLD analyzed represented an infectious mononucleosis-like (IM-like) lesion, 5 P-PTLD, and 6 M-PTLD. All P-PTLD and 5 M-PTLD had late-germinal center (GC)/post-GC phenotypes and 8 PTLD, including the IM-like lesion, were EBV+, while 4 PTLD were EBV−. The expression profiles of late GC/post GC PTLD appeared homogeneous and distinct from other B-NHL, however no segregation of P-PTLD and M-PTLD or EBV+ and EBV− PTLD was noted. Surprisingly, on supervised analysis, only 4 genes were expressed at higher levels in PTLD compared to B-NHL and normal B-cell subsets (BLIMP-1, XBP-1, IFITM3, PCMT1) and only 2 genes were differentially upregulated in EBV+ PTLD (CD27 ligand, MTDH). The expression profiles of PTLD appeared most similar to LCLs.Conclusions: B-cell PTLDs of late-GC/post-GC phenotype represent a distinct type of B-NHL, which are possibly derived from activated B-cells. Despite morphologic distinctions, P-PTLD and M-PTLD have overlapping genetic profiles suggesting disease heterogeneity within different categories.

Differential expression of the human MIXL1 gene product in non-Hodgkin and Hodgkin lymphomas

The Mix1 homeobox-like (MIXL1) gene encodes a paired class homeobox transcription factor that is involved in embryogenesis. Previous studies have shown that the MIXL1 gene product is expressed in B- and T-cell progenitors of normal bone marrow and, in some cell lines derived from hematopoietic neoplasms. The status of MIXL1 expression and subcellular localization in human lymphomas is unknown. Using a highly specific antibody, we assessed for MIXL1 expression in lymphoma cell lines of B- and T-cell lineage by reverse transcriptase-polymerase chain reaction, Western blot analysis, and immunohistochemistry. We also assessed for MIXL1 expression using immunohistochemical methods in 193 lymphoid tumors, including 140 B-cell non-Hodgkin lymphomas (NHL), 36 T-cell NHL, and 17 Hodgkin lymphomas (HL). MIXL1 was detected predominantly in the nuclear fraction of all cell lines tested and was predominantly nuclear in primary tumor specimens. Based on the distribution of the staining results (histogram), a 50% cutoff was selected for high versus low MIXL1 expression. High MIXL1 expression was detected more frequently in Burkitt lymphoma and diffuse large B-cell lymphoma compared with other types of B-cell NHL (P < .0001, chi(2) test). Most cases of T-cell NHL and all cases of HL also highly expressed MIXL1. Most plasma cell myelomas were negative for MIXL1, but rare cases had low MIXL1 expression. MIXL1 expression significantly correlated with proliferation index (Ki-67) in B-cell NHL (P < .0001). The frequent and high expression of MIXL1 in aggressive B-cell NHL, T-cell NHL, and HL suggests that MIXL1 may be involved in lymphomagenesis.

Preliminary Report of a Phase 1 Study of CMC-544, an Antibody-Targeted Chemotherapy Agent, in Patients with B-Cell Non-Hodgkin's Lymphoma (NHL).

Introduction: CMC-544 is an antibody-targeted chemotherapy agent composed of a monoclonal antibody, which specifically targets the CD22 antigen, conjugated to calicheamicin, a potent cytotoxic antitumor antibiotic. Malignant cells of mature B-lymphocyte lineage express CD22 and thus, CMC-544 may be useful for treating lymphomas of B-cell origin.Methods: A phase 1, dose escalation trial of CMC-544 is ongoing in patients (pts) with relapsed or refractory B-cell NHL across 13 European and US sites. CMC-544 was administered intravenously every 3–4 weeks at doses of 0.4, 0.8, 1.34, 1.8, and 2.4 mg/m2. Standard safety and pharmacokinetic data and preliminary efficacy data (assessed using the International Workshop to Standardize Response Criteria for NHL) are being collected. Enrollment of pts with any type of B-cell NHL, except for Burkitt's and lymphoblastic lymphomas, was allowed in the dose escalation phase (1–6 pts per cohort). After the maximum tolerated dose (MTD) was identified, 15 pts with follicular lymphoma and 15 pts with diffuse large B-cell lymphoma will be enrolled in an expanded MTD cohort.Results: As of June 2005, 34 pts (8 women, median 71 yrs; 26 men, median 62 yrs; number of previous treatments, median 4, range 2–11) were enrolled. Dose escalation was based on 1st cycle safety evaluations of pt cohorts. Dose-limiting toxicities (DLTs) were reported for dose levels of 1.34 mg/m2 (grade 4 thrombocytopenia, 2/11 pts), 1.8 mg/m2 (bleeding requiring platelet transfusion, 1/6), and 2.4 mg/m2 (grade 4 thrombocytopenia, 1/6; grade 4 neutropenia for 7 days, 1/6). Thus, the MTD, the dose level prior to the one where ≥ 33% DLTs occurred, was 1.8 mg/m2. The most common drug-related adverse events (AEs, all grades) were: thrombocytopenia (65%), asthenia (47%), nausea (41%), neutropenia (29%), elevated liver function tests (27%), anorexia (14%), and epistaxis (12%). Grade 3–4 AEs that occurred with a frequency ≥ 10% included: thrombocytopenia (38%), asthenia (12%), and neutropenia (12%). The nadir of the thrombocytopenia was 9±2 days and platelet counts recovered spontaneously. No major bleeding episodes were reported. At the 1.8 and 2.4 mg/m2 dose levels, data suggested a dose-dependent component of platelet recovery to baseline levels and dose delays were required between successive doses. CMC-544 and total calicheamicin exposures in serum increased with dose. Clearance after 2nd or 3rd doses decreased approximately 8–14-fold compared with 1st dose, and half-life increased from approximately 1 day to 4 days. Available data suggested an association of peak CMC-544 exposures and/or total calicheamicin levels with decreases in platelet counts. Accordingly, pts in the expanded MTD cohort receive 1.8 mg/m2 CMC-544 every 4 weeks. Preliminary antitumor activity was observed in most cohorts. Complete and/or partial responses were observed in the 0.8 mg/m2 (1/3 pts), 1.34 mg/m2 (3/9), 1.8 mg/m2 (2/5), and 2.4 mg/m2 (2/5) cohorts, and in the 1st 6 pts in the expanded MTD cohort (4/6).Conclusion: CMC-544 shows promising preliminary efficacy in pts with B-cell NHL, with an associated thrombocytopenia that is clinically manageable. The expanded MTD cohort (1.8 mg/m2 every 4 weeks) continues to be enrolled.

A Pilot Study of Yttrium 90 (90Y) Ibritumomab Tiuxetan (Zevalin®) in Patients with Relapsed and Refractory Classical Hodgkin Lymphoma (HL): Delivering Cross Fire Radiation by Targeting Reactive B Cells in the Microenvironment.

Both non-Hodgkin lymphoma (NHL) and HL are radiosensitive tumors. Although the efficacy of radioimmunotherapy (RIT) targeting CD20 antigen has been established in several types of B-cell NHL, no data exisit on the potential use of this approach in classical HL. The malignant Hodgkin and Reed-Sternberg (HRS) cells of HL infrequently express CD20 suggesting that classical HL may not be the optimal tumor to be targeted by this approach. However, HRS cells are frequently surrounded by reactive B lymphocytes that express CD20. Therefore, we hypothesized that targeting reactive B cells in HL microenvironment by RIT could deliver effective radiation dose to HL lymph nodes killing HRS cells by cross-fire effect. To test this hypothesis, we treated 4 patients with relapsed classical HL with Zevalin RIT. During the imaging phase, patients received a slow intravenous infusion of rituximab (250 mg/m2) followed by an intravenous injection of 4–5 mCi of In-111 labeled anti-CD 20 (In-111 Zevalin). Whole-body scans were performed at 2–4 hours and 40–48 hours after injection using a Siemens dual-head gamma camera. Simultaneous anterior and posterior projections were obtained at these time points. After ruling out any unfavorable biodistribution of Zevalin was confirmed on images at both time points, patients underwent the treatment phase which consisted of rituximab (250 mg/m2) followed by Y-90 anti-CD 20 (Y-90 Zevalin). To-date 4 patients are enrolled on study (ages 25, 31, 65, and 74 years). Two patients received prior ASCT, and 2 were treated after failing ABVD chemotherapy alone. All patients received 0.4 mCi/Kg up to a maximum dose of 32 mCi. Normal biodistribution of Zevalin was confirmed in all 4 patients during the imaging phase. No Zevalin-avid lesions was noted in one patient. There was faint visualization of some of the known lesions (by Ga-67 SPECT, CT or F-18 FDG PET) in the axilla and abdomen in 2 of the 4 patients. Intense Zevalin-uptake sites were identified in the supraclavicular, infraclavicular, mediastinal, hepatic and abdominal lymphatic chains in another patient. Imaging results and treatment responses are shown in the following table:Patient ResponsePrior ASCTIn-111 Zevalin-avidityClinical1YesnonePD2Nofaint, right axillaPD3Yesvery faint, abdomenNC4NointenseNCTreatment was well tolerated with hematologic toxicity similar to those observed in NHL patients. Our preliminary results suggest that RIT targeting reactive B cells in classical HL is feasible and safe. Future studies should incorporate pretreatment evaluation of a selected lymph node by fine needle aspiration to determine the fraction of reactive B cells in the microenvironment prior to treatment with RIT targeting CD20.

Clinicopathologic study of 369 B-cell non-Hodgkin lymphoma cases, with reference to the 2001 World Health Organization classification of lymphoid neoplasms

To describe the relative frequency, morphologic features, immunophenotype and clinical data of different types of B-cell non-Hodgkin lymphoma (B-NHL) and to evaluate the practical application of the 2001 World Health Organization (WHO) classification of lymphoid neoplasms. 369 documented cases of B-NHL were further subtyped according to the 2001 WHO classification of lymphoid neoplasms, on the basis of hematoxylin and eosin staining, immunohistochemistry and in-situ hybridization techniques. Amongst the 369 cases of B-NHL studied, 353 cases could be further classified into 11 subtypes. Diffuse large B-cell lymphoma, extranodal marginal zone lymphoma and follicular lymphoma were the commonest subtypes, accounting for 51.2% (189 cases), 14.9% (55 cases) and 10.6% (39 cases) of all cases respectively. Tumors in lymph nodes were seen in 158 cases (42.8%) and in extra node in 211 cases (57.2%). B-cell prolymphocytic leukemia and hairy cell leukemia were not identified. When comparing the diagnosis based on morphologic examination alone with the diagnosis based on both morphology and immunophenotype, there was a 80% concordance rate. Immunohistochemical study was helpful in reaching the correct diagnosis in many cases and could improve the overall diagnostic accuracy by about 20%. Amongst cases of B-NHL, diffuse large B-cell lymphoma is the commonest subtype, followed by MALToma and follicular lymphoma. While morphologic examination forms the basis for lymphoma diagnosis, immunohistochemical study also plays an important role in further subtyping. A combination of both modalities are sufficient for arriving at an accurate diagnosis in most cases of B-NHL, in keeping with the recommendation of the 2001 WHO classification of lymphoid neoplasms.

Aberrant Somatic Hypermutation Targets an Extensive Set of Genes in Diffuse Large B-Cell Lymphoma.

Genomic instability is a driving force in tumor development that can be achieved by a variety of mechanisms, such as defective chromosome segregation or inactivation of the DNA mismatch repair pathway. Although B-cell lymphomas are associated with chromosomal translocations deregulating oncogene expression, a mechanism for genome-wide instability during lymphomagenesis has long not been described. We have reported that the somatic hypermutation process (SHM), which normally targets the immunoglobulin variable region (IgV) and BCL6 genes in germinal center (GC) B-cells, functions aberrantly in >50% of diffuse large B-cell lymphoma (DLBCL), the most common type of B-cell non-Hodgkin lymphoma (Pasqualucci et al., Nature 412:341, 2001). As a consequence, multiple somatic mutations are introduced into the 5′ region of genes that do not represent physiologic SHM targets, including known proto-oncogenes such as PIM1, PAX5, RhoH/TTF and cMYC. To further define the extent of this phenomenon, termed aberrant somatic hypermutation (ASHM), and to identify additional hypermutated loci of possible pathogenetic significance in DLBCL, we screened 113 genes for the presence of mutations affecting their 5′ sequences (≥1.3 Kb from the transcription start site, the target region for SHM) in 10 DLBCL cell lines. Fifteen genes (13.3%) were found to harbor a significant number of mutations (p<0.05), with 70% of the cell lines being mutated in 7 or more genes; among these, six B-cell specific loci -BCL7A, CIITA, IRF4, LRMP, NCOA3 and SIAT1- carried 9–53 mutational events distributed in 20 to 70% of the cases, corresponding to an overall mutation frequency of 0.032–0.15% (frequency in the mutated cases: 0.07–0.25%). The same genes were found hypermutated in a panel of 20 primary DLBCL biopsies, which displayed an overall mutation load of 7 to 45 distinct events/gene (total N=125). Mutations were of somatic origin, independent of chromosomal translocations to the Ig loci and were restricted to the first 1.5–2 Kb from the promoter. In addition, analogous to previously identified SHM and ASHM targets, the mutations exhibited characteristic features, including a bias for transitions over transversions, preferential hotspot (RGYW/WRCY motifs) targeting, and higher frequencies at G:C pairs. However, in contrast to physiologic SHM targets such as IgV and BCL6, none of the 4 newly identified hypermutated genes that have been analyzed so far (BCL7A, CIITA, SIAT1, LRMP) displayed significant levels of mutations in purified normal GC B-cells as well as in other B-cell malignancies. This finding indicates that these genes represent aberrant hypermutation targets resulting from a tumor-associated malfunction, possibly a loss of target specificity of the physiologic SHM process. Considering previous results and the present survey, 17 (13%) out of 130 genes investigated have been found involved in ASHM, suggesting that this aberrant activity may involve an extensive set of target genes in DLBCL. Since the mutations affect both regulatory and coding sequences of the targeted genes, aberrant SHM may represent a major contributor to the pathogenesis of this disease and may explain in part its phenotypic and clinical heterogeneity.

A case of primary B-cell type Non-Hodgkin lymphoma originating in the iris

Purpose To report a case of primary malignant B-cell type non-Hodgkin lymphoma originating in the iris. Design Interventional case report. Methods An 83-year-old woman presented with anterior uveitis resulting from primary malignant lymphoma in the iris. Ultrasound biomicroscopy and indocyanine green angiography using a scanning laser ophthalmoscope showed abnormalities in the iris. Diagnostic biopsy of the iris revealed B-cell type non-Hodgkin lymphoma. Result The patient was treated with radiotherapy, and the tumorous lesion resolved. Conclusion Primary lymphoma localized in the iris only is rare. In this case, diagnostic biopsy and radiotherapy of the iris lymphoma provided good results.

Characterization of 4 Mantle Cell Lymphoma Cell Lines

Abstract Context.—Mantle cell lymphoma (MCL) is a distinct type of B-cell non-Hodgkin lymphoma characterized by t(11;14)(q13;q32) and cyclin D1 overexpression. The pathogenesis of MCL has not been comprehensively studied, which can be attributed in part to the paucity of well-characterized MCL cell lines. Objectives.—We collected 4 previously developed MCL cell lines and performed extensive characterization, including the susceptibly of these cell lines to transduction by adenovirus vectors. Our aim was to facilitate the establishment of an in vitro model that can be reliably used to study the pathogenesis of MCL. Methods.—Standard techniques were used to compare the morphologic, immunophenotypic, and cytogenetic features of the 4 cell lines. In addition, Western blotting was used to investigate the presence of several cell cycle- and apoptosis-related proteins. TP53 DNA sequencing was also performed on the cell lines. The adenoviral transduction efficiency was assessed using an adenoviral vector carrying...

Characterization of 4 mantle cell lymphoma cell lines.

Mantle cell lymphoma (MCL) is a distinct type of B-cell non-Hodgkin lymphoma characterized by t(11;14)(q13;q32) and cyclin D1 overexpression. The pathogenesis of MCL has not been comprehensively studied, which can be attributed in part to the paucity of well-characterized MCL cell lines. We collected 4 previously developed MCL cell lines and performed extensive characterization, including the susceptibly of these cell lines to transduction by adenovirus vectors. Our aim was to facilitate the establishment of an in vitro model that can be reliably used to study the pathogenesis of MCL. Standard techniques were used to compare the morphologic, immunophenotypic, and cytogenetic features of the 4 cell lines. In addition, Western blotting was used to investigate the presence of several cell cycle- and apoptosis-related proteins. TP53 DNA sequencing was also performed on the cell lines. The adenoviral transduction efficiency was assessed using an adenoviral vector carrying the gene encoding for the green fluorescence protein (Ad-GFP). All cell lines demonstrated evidence of t(11;14)(q13;q32) and overexpression of cyclin D1. Cyclin D2 was not detectable in all cell lines, whereas cyclin D3 was weakly expressed in JeKo-1 and SP-53. Other abnormalities of the cell cycle G1 phase regulatory pathway were detected, including loss of expression of p53 (JeKo-1) and p16(INK4a) (SP-53 and Granta 519), as well as TP53 mutation (Mino). All cell lines express high levels of cyclin E, c-Myc, Bcl-2, Bax, Bcl-x(L), and Mcl-1. Retinoblastoma protein is hyperphosphorylated in all cell lines. With the exception of Mino, MCL cell lines are highly transducible with adenoviral vectors. These cell lines are representative of MCL and can be used as an in vitro model to further explore the pathogenesis of this disease. The susceptibility of these cell lines to gene transfer provides opportunities to evaluate the importance of various oncogenes and tumor suppressor genes that may have an impact on developing effective therapeutic regimens for MCL.

Monoclonal antibody FMC7 detects a conformational epitope on the CD20 molecule: evidence from phenotyping after rituxan therapy and transfectant cell analyses.

Numerous studies have reported that monoclonal antibody (mAb) FMC7 detects an antigen present on only a subset of circulating B lymphocytes. In particular, this mAb may distinguish typical B-cell chronic lymphocytic leukemia (FMC7 negative) from other types of B-cell non-Hodgkin lymphoma (B-NHL; FMC7 positive). We treated patients with B-NHL with Rituxan, a chimeric CD20 mAb, and observed abrogation of staining not only with prototype CD20 mAb B-1 but also with mAb FMC7. To investigate the relation between antigens CD20 and FMC7, we performed mutual blocking studies that showed mutual inhibition of FMC7 and CD20. In addition, FMC7 modulated CD23 expression and confirmed the presence of mAb B-1 in B-lymphoblastoid cell lines CESS and JVM. Transient transfection of myeloid cell line K562 with plasmid containing CD20-encoding cDNA produced de novo expressions of CD20 and FMC7. Our data indicate that FMC7 binds to a particular conformation of the CD20 antigen, probably to a multimeric CD20 complex. We assume that FMC7 stains positively only when CD20 antigen is present in high densities and in the postulated multimeric complex formation.

Telomerase activity in B-cell non-Hodgkin lymphoma.

Studies have shown telomerase activity to be present in some B-cell non-Hodgkin lymphomas (B-NHLs). However, no large studies have assayed telomerase activity in a systematic and quantitative manner. Furthermore, the relation between telomerase and proliferation suggested by in vitro studies has not been adequately tested in B-NHLs in vivo. This information is necessary to understand the relation between proliferation and telomerase and to predict the efficacy of antitelomerase drugs currently in development. Eighteen benign biopsies and 111 B-NHLs of varying types were classified according to the revised European-American classification of lymphoid neoplasms (REAL classification) and assayed for telomerase activity and proliferation index (PI). All B-NHLs contained telomerase activity except for low grade marginal zone B-cell lymphomas (MZBCLs) (96 of 111, 86%) (chi(2) 95.90, P < 0.001). Telomerase activity correlated with PI (r = 0.7536, r(2) = 0.5678, t = 10.51, P < 0.001) and showed a threshold whereby telomerase activity was not present below a PI of 9.2% (t = 4.875, P < 0.001). The level of telomerase activity fell within characteristic ranges and generally correlated with the clinical aggressiveness of each B-NHL category. Low grade MZBCLs of extranodal, nodal, and splenic types were unique among the categories of B-NHL in lacking or containing very little telomerase activity. The association between telomerase activity and PI is evidence that telomerase is controlled in vivo along with the cell cycle and is not constitutively active in B-NHL. These data provide evidence that antitelomerase drugs may be efficacious in most types of B-NHL.

Non-Hodgkin's Lymphoma Associated with Gaucher's Disease

Gaucher's disease is an uncommon disorder which has been reported to be associated with an increased risk of lymphoproliferative disorders, including Non-Hodgkin's lymphoma (NHL). A new instance of such an association is described here. This was a 58 year-old-patient with adult type I Gaucher's disease who, one and a half year after the above diagnosis, presented with supraclavicular lymphadenopathy, massive splenomegaly, prominent retroperitoneal lymphadenopathy and increased serum LDH levels. This led to the diagnosis of large-cell NHL of B-cell type, successfully treated with chemotherapy. The previously published cases of Gaucher's disease associated with NHL as well as the possible mechanisms leading to this association are reviewed here.

Prognostic significance of BCL-2 expression and bcl-2 major breakpoint region rearrangement in diffuse large cell non-Hodgkin's lymphoma: a British National Lymphoma Investigation Study

The Bcl-2 protein is capable of preventing apoptosis, and in vitro evidence suggests a role in drug resistance. It is expressed and the gene is rearranged in a proportion of cases of large-cell non-Hodgkin's lymphoma (NHL), but the clinical significance of these findings is controversial. The purpose of this study was to determine the influence of both Bcl-2 expression and major breakpoint region (MBR) bcl-2 rearrangement in a large cohort of prospectively accrued patients with intermediate-grade B-cell NHL treated in a standardized manner. All patients with Working Formulation F, G, or H NHL treated with cyclophosphamide, doxorubicin, vincristine, and prednisone (CHOP) chemotherapy in British National Lymphoma investigation studies between July 1974 and April 1992 were considered for this study if the appropriate paraffin blocks were available. Paraffin sections from the diagnostic specimen were analyzed for evidence of MBR rearrangement using a polymerase chain reaction-based method, and for Bcl-2 expression using immunohistochemistry. Failure to achieve complete remission (CR), relapse, death from NHL, and deaths from all causes were used as end points to measure CR rate, actuarial relapse rate, actuarial survival from NHL, and actuarial overall survival. One hundred sixty-one suitable patients were identified and tested for the bcl-2 MBR translocation, with 27 (17%) found to be positive; 153 of these patients were tested with immunocytochemistry, and 84 (55%) showed evidence of Bcl-2 expression. For patients who achieved CR from the initial treatment, the relapse rate was significantly higher in those with Bcl-2 expression than in those without. In addition, multivariate analysis identified Bcl-2 expression as the only factor significantly related to relapse rate in the subjects measured. The cause-specific survival for NHL in the series as a whole was significantly lower in patients with Bcl-2 expression than in those without. MBR status had no significant influence on any of the outcome measures, but the number of MBR-positive patients was relatively small, and larger studies are required. In conclusion, in Working Formulation F, G, and H NHL of B-cell type, expression of Bcl-2 protein predicted independently for relapse.

Prognostic Significance of BCL-2 Expression and bcl-2 Major Breakpoint Region Rearrangement in Diffuse Large Cell Non-Hodgkin's Lymphoma: A British National Lymphoma Investigation Study

The Bcl-2 protein is capable of preventing apoptosis, and in vitro evidence suggests a role in drug resistance. It is expressed and the gene is rearranged in a proportion of cases of large-cell non-Hodgkin's lymphoma (NHL), but the clinical significance of these findings is controversial. The purpose of this study was to determine the influence of both Bcl-2 expression and major breakpoint region (MBR) bcl-2 rearrangement in a large cohort of prospectively accrued patients with intermediate-grade B-cell NHL treated in a standardized manner. All patients with Working Formulation F, G, or H NHL treated with cyclophosphamide, doxorubicin, vincristine, and prednisone (CHOP) chemotherapy in British National Lymphoma investigation studies between July 1974 and April 1992 were considered for this study if the appropriate paraffin blocks were available. Paraffin sections from the diagnostic specimen were analyzed for evidence of MBR rearrangement using a polymerase chain reaction-based method, and for Bcl-2 expression using immunohistochemistry. Failure to achieve complete remission (CR), relapse, death from NHL, and deaths from all causes were used as end points to measure CR rate, actuarial relapse rate, actuarial survival from NHL, and actuarial overall survival. One hundred sixty-one suitable patients were identified and tested for the bcl-2 MBR translocation, with 27 (17%) found to be positive; 153 of these patients were tested with immunocytochemistry, and 84 (55%) showed evidence of Bcl-2 expression. For patients who achieved CR from the initial treatment, the relapse rate was significantly higher in those with Bcl-2 expression than in those without. In addition, multivariate analysis identified Bcl-2 expression as the only factor significantly related to relapse rate in the subjects measured. The cause-specific survival for NHL in the series as a whole was significantly lower in patients with Bcl-2 expression than in those without. MBR status had no significant influence on any of the outcome measures, but the number of MBR-positive patients was relatively small, and larger studies are required. In conclusion, in Working Formulation F, G, and H NHL of B-cell type, expression of Bcl-2 protein predicted independently for relapse.

Detecting of the minimal residual disease contaminated in peripheral blood stem cell transplantation in the B-cell malignant lymphoma patients.

For sufficient collection of hemopoietic stem cells from peripheral blood for autologous peripheral blood stem cell transplantation (PBSCT), four patients with B-cell-type non-Hodgkin lymphoma (B-NHL) were examined for the appearance of circulating hemopoietic progenitors in blood (PSC) during the hemopoietic recovery phase following marrow ablative therapy in combination with or without administration of recombinant human granulocyte colony-stimulating factor (rhG-CSF). Each patient received only chemotherapy in the first course, and rhG-CSF (1 microgram/kg/day) was administered for 14 consecutive days from the last day of the second chemotherapy. In the second chemotherapy course with rhG-CSF administration, white blood cell (WBC) counts demonstrated two peaks, and the appearance of granulocyte-macrophage precursor cells (CFU-GM) in blood at the maximum level was coincident with the second peak of WBC elevation. Erythroid precursor cells (BFU-E) were also detectable in blood after chemotherapy but the peak level was not enhanced by the use of rhG-CSF. To determine whether the minimal residual disease (MRD) cells were contaminated in PSC corrected from blood, kappa-lambda imaging (KLI) analysis was performed to detect the malignant B-cell population (mBp) before and after chemotherapy. No mBp was found in two of four patients in blood, although three of them were involved with mBp in bone marrow. The presence of mBp was detected in two patients both before and after chemotherapy, even though these cells were hardly detected morphologically, suggesting the necessity of judging for the incidence of contamination of MRD cells when collecting PSCs.

Development of Non-Hodgkin Lymphoma in a Cohort of Patients with Severe Human Immunodeficiency Virus (HIV) Infection on Long-Term Antiretroviral Therapy

To describe the incidence of non-Hodgkin lymphoma in a group of patients with symptomatic human immunodeficiency virus (HIV) infection receiving long-term dideoxynucleoside antiretroviral therapy. We examined the records of all patients with the acquired immunodeficiency syndrome (AIDS) or severe AIDS-related complex who were entered into three long-term phase I trials of zidovudine (azidothymidine, AZT) or zidovudine-containing regimens at the National Cancer Institute between 1985 and 1987. The Warren G. Magnuson Clinical Center, National Institutes of Health, Bethesda, Maryland. Fifty-five HIV-infected patients with AIDS or severe AIDS-related complex. Eight of fifty-five patients (14.5%; 95% CI, 6.5% to 26.7%) developed a high-grade non-Hodgkin lymphoma of B-cell type, a median of 23.8 months (range, 13 to 35 months) after starting antiretroviral treatment. Using the method of Kaplan and Meier, the estimated probability of developing lymphoma by 30 months of therapy was 28.6% (CI, 13.7% to 50.3%) and by 36 months, 46.4% (CI, 19.6% to 75.5%). The patients who developed lymphoma had less than 100 T4 cells/mm3 for a median of 17.8 months (range, 7 to 35 months) and less than 50 T4 cells/mm3 for a median of 15.3 months (range, 5.5 to 35 months) before the diagnosis. All patients presented with non-Hodgkin lymphoma in extranodal sites, and two developed primary brain involvement in the setting of Toxoplasma infection. Patients with symptomatic HIV infection who survive for up to 3 years on antiretroviral therapy may have a relatively high probability of developing non-Hodgkin lymphoma. Prolonged survival in the setting of profound immunosuppression with substantial T4-cell depletion is probably an important factor in the development of these lymphomas. However, a direct role of therapy itself cannot be totally discounted. As improved therapies for the treatment of HIV infection and its complications result in prolonged survival, non-Hodgkin lymphoma may become an increasingly significant problem.