A novel B cell line established from Ki-1-positive diffuse large cell lymphoma

A novel cell line, designated KIS-1, was established from a patient with Ki-1-positive diffuse large cell lymphoma. Multiple phenotypic analysis of the KIS-1 cells was carried out with a total of 22 monoclonal antibodies defining hematopoietic cell subsets and lineages. The KIS-1 cells were positive for Ki-1, B4, HLA-DR, and 2D1 (common leucocyte) antigens, but were negative for the antigens reportedly specific for T cells, natural killer cells, granulocytes, monocytes, interdigitating reticulum cells and dendritic reticulum cells. The genomic analysis of the KIS-1 cells showed not only the rearrangement of JH and J kappa genes but also the probable rearrangement of C lambda genes. Moreover, the cells produced immunoglobulin lambda chains. Thus, KIS-1 was considered to be of B-cell lineage. The lymphoma-cell derivation of KIS-1 was based on the following facts. The cytochemical, immunologic, cytogenetic properties and the results of the molecular genomic analysis in the KIS-1 cells were essentially the same as those of the original tumor cells, and the KIS-1 cells were negative for Epstein-Barr virus-associated nuclear antigen. KIS-1 is the only known B-cell line derived from Ki-1-positive diffuse large cell lymphoma, and should be useful for defining the biological implications of Ki-1 antigen.     

Establishment of a novel cell line with T-lineage phenotype (HPB-MLp-W) from a non-Hodgkin's lymphoma patient

We report the characterization of a novel human T-cell line, HPB-MLp-W, which was established from blastic cells of a lymph node specimen from a patient with non-Hodgkin's lymphoma. They demonstrated the T-cell association antigens, CD2 and CD4, but no CD3, CD8, CD1, CD5, CD7 nor T-cell antigen receptor on their cell surfaces. They were also positive for Ia and Ki-1 antigen, and negative for CD25 (Tac-1). The cell line HPB-MLp-W had the same pattern of antigen expression as the patient's cells. Southern-blot analysis of DNA showed a rearrangement of the T-cell receptor-alpha and beta genes. To our knowledge, this is a novel cell line with unique T-lineage marker, to be established from a case of non-Hodgkin's lymphoma.     

t(2;5) (p23;q35)-A Specific Chromosome Abnormality in Large Cell Anaplastic (Ki-1) Lymphoma

Chromosome abnormalities of three patients with Ki-1 lymphoma are presented. In order to be included in the study each case fulfilled the following criteria: the majority of the tumour cells were positive for the Ki-1 antigen (CD30), and the cells were large with relatively abundant, slightly basophilic cytoplasm. In all cases, a major proportion of mitoses contained a complicated clonal chromosome abnormality. Two patients had a 2;5 translocation; and the third had break points at 14q32 and 2p12. The latter patient showed expression of B-cell antigens and had rearrangements in the immunoglobulin heavy chain and kappa light chain genes. The two patients with the 2;5 translocation were epithelial membrane antigen positive, but did not exhibit rearrangements of immunoglobulin/T-cell receptor genes or expression of T-/B-cell antigens.     

Establishment of a novel B-lymphoma cell line, CTB-1, with strong Pas antigen expression having chromosomal translocation (14;22)

We established a new lymphoma cell line, designated CTB-1, from pericardial effusion of a patient with diffuse large B-cell lymphoma. This cell line showing vigorous growth ability has undergone 260 passages over a period of 34 months in suspension culture, and is heterotransplantable to nude mice. The cultured cells were positive for CD10, CD19, CD20, CD21, HLA-DR, and surface IgG kappa, and negative for T cell antigens. Chromosomal analysis revealed a t(14;22)(q32;q11) that is consistent with original lymphoma cells. CTB-1 cells show the high cell surface expression level of Fas antigen/APO-1. However, ligation of Fas antigen with anti-Fas monoclonal antibody (clone CH-11) did not induce apoptosis of CTB-1 cells. This suggests that Fas itself or the downstream signaling pathways of Fas may be impaired in this cell line. This new cell line may provide a useful in vitro system to study the biology and pathogenesis of B-cell lymphoma which is independent of Fas-mediated apoptosis.     

The prognostic value of Ki-67 antigen in non-Hodgkin lymphoma of Waldeyer ring and the nasal cavity.

BACKGROUND. A monoclonal antibody, Ki-67, recognizes an antigen expressed in all phases of the cell cycle, except G0, and can be used as a simple histologic marker of cell proliferation. To assess the prognostic value of the growth fraction in non-Hodgkin lymphoma of Waldeyer ring (W-NHL) and the nasal cavity (N-NHL), the authors applied Ki-67 immunostaining combined with image analysis on such lymphomas. METHODS. The authors studied 29 patients (18 with W-NHL and 11 with N-NHL), applying Ki-67 to frozen sections. The number of Ki-67-positive cells in a unit area (0.044 mm2), as an indicator of proliferative activity, and the mean area per Ki-67-positive cell (microns2), as an indicator of DNA content, were measured by the image processing system. RESULTS. High-grade lymphomas showed a significantly larger number of Ki-67-positive cells than intermediate-grade lymphomas (102.5 +/- 21.6 in high-grade and 46.8 +/- 8.92 in intermediate-grade lymphomas, P = 0.03), even when analyzed separately by immunophenotypes. A large mean area per Ki-67-positive cell was associated significantly with a T-cell phenotype (36.3 +/- 7.69 microns2 in T-cell lymphomas and 19.4 +/- 2.33 microns2 in B-cell lymphomas, P = 0.034) and an unfavorable clinical outcome. High proliferative activity, defined as nuclear Ki-67 expression in 2000 or more B-cell lymphoma cells and 1000 or more T-cell lymphoma cells in a 1-mm2 area, was found to be a strong predictor of poor survival among these patients (P = 0.048 and P = 0.009, respectively). CONCLUSIONS. Ki-67 immunostaining, combined with image analysis, is a novel method for determining a tumor proliferative index that provides useful clinical data regarding head and neck lymphomas.     

Long-term survival in Ki-1 lymphoma

Three patients with histologic and immunologic features of Ki-1-positive large cell lymphoma, who experienced long-term survival, are presented. These three patients at 2, 28, and 49 years of age had adenopathy; all cases had been initially misdiagnosed as metastatic carcinoma or malignant histiocytosis. On subsequent review, they had sinusal and diffuse growth of large pleomorphic cells that were Ber-H2 (Ki-1; CD 30) positive. One case marked as a T-cell lymphoma with UCHL1, one case expressed T-cell and B-cell markers, and one case was negative for both T-cell and B-cell markers. All patients received chemotherapy, and two received local radiation. One patient was not treated until 9 years after initial diagnosis. Two patients had several recurrences, but there has been no evidence of lymphoma in any of the three patients for 63 to 301 months; overall survival time has ranged from 14 to 25 years. These cases are the longest reported survivors with Ki-1 lymphoma; 5 years was the longest survival time previously reported. It also is noteworthy that Ber-H2 and other lymphoid-associated antigens appear to be preserved in formalin-fixed, paraffin-embedded tissues for prolonged periods. This may allow retrospective studies to evaluate the natural history of Ki-1 lymphomas, as well as their spontaneous or treatment-induced regression.     

A Ki-1 (CD30)-positive T (E+, CD4+, Ia+)-cell line, DL-40, established from aggressive large cell lymphoma

A new human lymphoma cell line, designated DL-40, was established from the peripheral blood of a 64-year-old woman with leukemic conversion of aggressive large cell lymphoma. The cell line grew in suspension with or without forming clumps of cells and exhibited large, round, or multiple nuclei in the relatively abundant cytoplasm that was positive for acid phosphatase. The cells expressed a Ki-1 antigen (CD30), E+, CD2+, CD4+, CD45+, Ia+ phenotype and had rearranged T-cell receptor beta chain but were negative for CD15, HTLV-I, and Epstein-Barr virus nuclear antigen. Chromosome analysis of this cell line showed a human female karyotype with complex hyperdiploid abnormalities. DL-40 cells produced tumors histologically similar to the original lymphoma when transplanted into nude mice and immunosuppressed hamsters. The DL-40 cell line could provide a useful tool for the understanding of biology of the Ki-1-positive non-Hodgkin's lymphoma.     

Anaplastic Ki-1-positive large cell lymphoma of the pancreas: a case report and review of the literature

A case of Ki-1-positive anaplastic large cell lymphoma of the pancreas is presented. The patient complained of abdominal pain and was jaundiced. Examination of a biopsy specimen obtained by duodenal endoscopy revealed malignant lymphoma, and surgery confirmed a large mass located in the region from the intra-pancreatic tissue around the lower common bile duct to the peri-pancreatic lymph nodes. Histologically, this tumor was composed mainly of large and giant neoplastic cells. Immunohistochemically, these cells were diffusely positive for Ki-1 and CD45RO antigens, indicating the features of Ki-1 anaplastic large cell lymphoma with a T-cell phenotype among non-Hodgkin's lymphoma. The histologic types of the majority of malignant lymphomas of the pancreas reported previously were considered to be diffuse-type non-Hodgkin lymphoma (probably with predominance of the B-cell phenotype), except for a single Japanese lymphoma case with a T-cell phenotype. This is therefore the first known case of Ki-1 anaplastic large cell lymphoma of the pancreas.      

Immunoglobulin Somatic Variation; Studies of Receptor Editing in a Human B Cell Lymphoma

In order to study mechanisms of immunoglobulin somatic variation in committed immunoglobulin (Ig) expressing B cells, we used the fluorescent activated cell sorter to isolate rare variants from a surface Ig positive (sIg⁺) diffuse large cell B lymphoma cell line (μλ⁺). These variants were either negative for sIg expression (sIg^-) or expressed sIg which differed from the original parental tumor cell line, both in idiotypes and Igλ isotypes (sIg⁺Id^-). In the following report we review the results from the studies of these variants. DNA analysis showed that all variants had new Igλ gene rearrangements, which had occurred either on a previously excluded allele, or on the productively rearranged allele of the parental cell line. The slg^- variants, which had undergone nonfunctional Igλ rearrangements on the expressed parental allele, and thereby deleted the productive rearrangement, continued to functionally rearrange the same allele and regenerated sIg expression. While the parental cell line expressed low levels of the recombination activating genes, RAG1 and RAG2, expression of these genes were considerably upregulated in both the immunoglobulin negative and the idiotypic variants. Somatic immunoglobulin V gene hypermutation did not contribute to the observed immunoglobulin somatic variation. These results demonstrate that, through differential expression of the RAG genes, sIg⁺ B cells are able to somatically alter their sIg receptors through secondary Igλ gene rearrangements. This mechanism may allow B cells with non-selectable, or auto-reactive, antigen receptors to alter these receptors (receptor editing). Ongoing Ig gene rearrangement may limit the usefulness of immunotherapeutic approaches directed at the antigen receptor in some diffuse large cell lymphomas.     

Establishment of Epstein-Barr virus-negative diffuse large cell lymphoma cell line with an 8;22 chromosomal translocation

A new human B-cell lymphoma cell line was established from a pleural effusion of a patient with a diffuse large cell lymphoma which originated from an ileocecal tumor. The cell line, designated KAL-1, has been passaged 280 times over a period of 22 months. This cell line was successfully maintained in a chemically defined serum-free medium; its doubling time is approximately 24 h. Immunologically, the cells were demonstrated to express IgM lambda on the cell surface and to react with monoclonal antibodies to B-cell antigen including B1, B4, HLA-DR, and common acute lymphoblastic leukemic antigen but not with B2 and all the T-cell markers. Immunoglobulin gene analysis revealed rearrangements of both JH and C lambda. These data indicate that this cell line represents the B-cell lineage at the immature B-cell stage. This cell line was negative for Epstein-Barr virus nuclear antigen and had no detectable Epstein-Barr virus genome in cellular DNA. Chromosome analyses revealed that the cells carried an 8;22 chromosome translocation, reminiscent of variant type Burkitt's lymphoma. However, there was no histological evidence for Burkitt's lymphoma. Molecular studies showed that KAL-1 had deregulated high constitutive expression of c-myc. This cell line was demonstrated to be highly tumorigenic when injected into athymic nude mice. This tumor model should provide clues about the molecular mechanism involved in the pathogenesis of B-cell malignancy and appears to be a useful in vivo model for the study of molecular events during B-cell differentiation and therapeutic investigations.     

Establishment of a human cell line (SKI-DLCL-1) with a t(1;14)(q21;q32) translocation from the ascites of a patient with diffuse large cell lymphoma.

Cytogenetic abnormalities at chromosome 1q21 are among the most common second genetic events observed in Non-Hodgkin's Lymphomas and have prognostic significance. Recently, BCL9 has been cloned from a pre-B-cell lymphoblastic leukemia cell line, which carried a t(1:14)(q21;q32). However, among a panel of 39 B-cell malignancies with 1q21 translocation, only two cases showed rearrangement for the BCL9 gene. We report the establishment of a new lymphoma cell line from a patient with relapsed diffuse large cell lymphoma. This cell line SKI-DLCL-1 showed cell surface antigens identical to the original tumor and demonstrated the profile of a mature B-cell phenotype: CD19 and CD20 positive, CD5 and C10 negative. It carried a t(1;14)(q21;q32) translocation identical to the original tumor. Although the clinical presentation was an isolated effusion lymphoma, studies for HIV-1, HHV8 and EBV were all negative. Southern blot analysis demonstrated that BCL9 was not rearranged in the SKI-DLCL-1 cell line. In addition, the BCL9 gene was not over-expressed in SKI-DLCL-1 cell line. The identification of a new locus at 1q21 will help clarify the pathogenesis of B-cell malignancies with a translocation involving this locus.     

Rearrangement of immunoglobulin heavy chain genes and t3 expression in the absence of rearrangement of t-cell receptor β-chain gene in a patient with t-cell malignant lymphoma

We describe the rearrangement of immunoglobulin genes and T3 expression in the absence of rearrangement of T-cell receptor β-chain genes in a patient wih T-cell malignant lymphoma. He had a mediastinal mass and his lymphoma cells expressed T-cell antigens (OKT3+, OKT9+, and OKT10+). When we examined genomic DNA from the lymphoma cells, we detected the rearrangement of immunoglobulin heavy chain genes with a germ-line configuration of light chain genes and no rearrangement of T-cell receptor β-chain gene. These results indicated that the rearrangement of immunoglobulin genes could occur in T-cell malignant lymphoma, and that T3 antigen could be expressed prior to the rearrangement of T-cell receptor β-chain genes under certain circumstances.     

Detection of Epstein-Barr virus DNA and expression of CD30 antigen in primary anaplastic diffuse large B-cell lymphoma of the brain

We describe a case of primary anaplastic diffuse large-cell lymphoma arising in the central nervous system (CNS). Primary CD30-positive anaplastic diffuse large B-cell lymphoma of the brain is very rarely reported. Given that this tumor is immunohistochemically heterogeneous, polymerase chain reaction (PCR) and Epstein-Barr virus (EBV) analysis of tumor DNA are essential techniques for early and accurate histological diagnosis in these CD30-positive cerebral lymphoma cases. We report an early CD30- and EBV-positive anaplastic diffuse large B-cell lymphoma in the CNS that was diagnosed not only from the immunohistochemical study and MRI findings, but also from the genotype confirmations. This tumor was documented to have EBV episomes of monoclonal origin by PCR analysis of immunoglobulin gene rearrangement.     

Inhibition of Natural Killer Cell Cytotoxicity by Cell Growth-related Molecules

Certain MHC class I molecules on target cells are known to inhibit the cytotoxic action of NK cells. By using monoclonal antibody (mAb) Cho-1, we have found inhibitory non-MHC class I cell surface molecules that are noncovalently-associated with 200 kDa and 40 kDa antigens. Poly I-C-induced rat NK cells were not cytotoxic to rat fetus-derived fibroblast WFB cell line. In contrast, NK cells were cytotoxic to H- ras oncogene-induced transformants of WFB, W14 and W31. FACS analysis indicated that mAb Cho-1 reacts with WFB, but not with W14 and W31 cells. Thus, this antigen may disappear concomitantly with cell growth and transformation. Cho-1 antigens were also expressed on other NK-resistant lines, such as mouse BALB3T3 fibroblast, EL-4 lymphoma and human fibroblast HEPM. However, they were not expressed on NK-sensitive mouse YAC-1 and H- ras transformant (Brash) of BALB3T3 cells. Furthermore, treatment of target cells with IFN-γ clearly induced the cell surface expression of Cho-1 antigens, and conferred a resistance to NK cytolysis on target cells. These data strongly suggest that Cho-I antigen expression may correlate with target cell susceptibility to NK cells. Indeed, treatment of NK-resistant WFB as well as HEPM cells with F(ab')₂ fragments of mAb Cho-1 resulted in the acquisition of susceptibility to NK cytolysis. Cho-1 antigens may be novel molecules that regulate the NK resistance of cells.     

A Multiparametric Study of Malignant Lymphoma of Mucosa Associated Lymphoid Tissue (Malt)

A morphological, immunophenotypic and ultrastructural study, cell cycle estimation, DNA and cytogenetic analysis were performed in ten cases of B-MALT lymphomas. Five had low grade lymphoma and five had high grade. Low and high grade cases showed the same cells but in different percentages: These included centrocyte-like cells with occasional monocytoid cytoplasmic changes, and centroblast-like cells. However, in high grade cases more dysplastic and large cells were present. All cellular types showed an important development of rough endoplasmic reticulum. In all cases a large panel of monoclonal antibodies was employed to study the B-cell immunophenotype. Ki-67 positivity ranged from 5% to 30% in low-grade cases and from 50% to 70% in high-grade cases. Gene rearrangement analysis showed rearrangement with Jh probe and half of the cases were also rearranged with the Kde probe (Kappa constant chain gene). A rearrangement banding pattern with TCR genes was not present in any of the cases. Cytogenetic study showed complex alterations in high grade cases and a normal karyotype in low grade lymphomas. Only one case had rearrangement for the bcl-2 probe.     

A multiparametric study of malignant lymphoma of mucosa associated lymphoid tissue (MALT).

A morphological, immunophenotypic and ultrastructural study, cell cycle estimation, DNA and cytogenetic analysis were performed in ten cases of B-MALT lymphomas. Five had low grade lymphoma and five had high grade. Low and high grade cases showed the same cells but in different percentages: These included centrocyte-like cells with occasional monocytoid cytoplasmic changes, and centroblast-like cells. However, in high grade cases more dysplastic and large cells were present. All cellular types showed an important development of rough endoplasmic reticulum. In all cases a large panel of monoclonal antibodies was employed to study the B-cell immunophenotype. Ki-67 positivity ranged from 5% to 30% in low-grade cases and from 50% to 70% in high-grade cases. Gene rearrangement analysis showed rearrangement with Jh probe and half of the cases were also rearranged with the Kde probe (Kappa constant chain gene). A rearrangement banding pattern with TCR genes was not present in any of the cases. Cytogenetic study showed complex alterations in high grade cases and a normal karyotype in low grade lymphomas. Only one case had rearrangement for the bcl-2 probe.     

U-2940, a human B-cell line derived from a diffuse large cell lymphoma sequential to Hodgkin lymphoma

Several patterns of association between Hodgkin and non-Hodgkin lymphomas are recognized, some of which support a common cellular origin or shared transformation events for both malignancies. We describe the U-2940 cell line derived from a diffuse large B-cell lymphoma with some features consistent with mediastinal large B-cell lymphoma, clinically apparent 1 month after the initial course of chemotherapy for Hodgkin's disease, fulfilling the criteria for composite malignancies. U-2940 cells display a mature B phenotype with hypermutated IgH rearrangement typical of germinal/postgerminal center origin. The cell line is negative for Epstein-Barr virus and no evidence of t(14;18) was found. U-2940 cells display multiple chromosomal rearrangements similar to recurrent aberrations described in both Hodgkin and non-Hodgkin lymphomas, also partially shared by U-2932 derived from a B-cell lymphoma sequential to Hodgkin's disease. The original large B-cell lymphoma and the U-2940 cell line bear microsatellite instability, an abnormality associated with particular subtypes of non-Hodgkin lymphomas and found in tissues involved by Hodgkin lymphoma. Therefore, U-2940 cells bear several features known to occur in Hodgkin and in non-Hodgkin lymphomas, leading to the assumption that this cell line may constitute a useful tool to address elective pathways of lymphomagenesis and eventually the Hodgkin and non-Hodgkin lymphoma association.     

CD20阳性血管免疫母细胞性 T 细胞淋巴瘤1例及文献复习

目的:探讨CD20阳性血管免疫母细胞性T细胞淋巴瘤（angioimmunoblastic T cell lymphoma,AITL）的临床特征及预后。方法:回顾分析我院1例CD20阳性AITL患者的临床病理特征、治疗转归并复习相关文献。结果:患者男性,69岁,以水肿及腹腔积液为首发表现,CT提示全身淋巴结肿大。免疫表型:CD20阳性、CD3(+)、CD5(+)、Ki-67(+,85%),其它B细胞标记阴性,EBER原位杂交阳性,TCR基因重排及IGH单克隆性重排阳性,多种治疗方案均无效。结论:CD20阳性AITL患者的临床病理特征易与B细胞淋巴瘤混淆,病理形态学、免疫组织化学及TCR基因重排检测等可减少误诊。利妥昔单抗及其他靶向药物的应用可能提高治愈率,改善患者预后。     

Phenotypic and genotypic lineage switch of a lymphoma with shared chromosome translocation and T-cell receptor gamma gene rearrangement.

A case of non-Hodgkin's lymphoma showed a phenotypic and genotypic cell lineage switch twice during nine years of his clinical history; first, T-cell type, pleomorphic small cell lymphoma developed, followed by B-cell type, diffuse centroblastic/centrocytic lymphoma, and finally T-zone lymphoma without follicles again developed, from which AST-1 cultured cell line was established. Karyotype analysis demonstrated a shared abnormal chromosome, der(1)t(1;?)(p36;?), among the first relapsed B-cell tumor, the second relapsed T-cell tumor and AST-1 cell line. Furthermore, T-cell receptor (TCR) gamma gene rearrangement bands of the same size were observed in the first relapsed B-cell tumor and the second relapsed T-cell tumor as well as AST-1 cell line. These results suggested that both relapsed tumors of different cell lineages are derived from a common malignant clone, presumably a committed lymphoid stem cell. A unique translocation, t(2;14)(q37;q11.2), which may involve TCR delta/alpha gene complex, was observed in the second relapsed tumor and AST-1 cells. To attempt to isolate the breakpoint of this translocation, the configuration of TCR delta/alpha gene complex was studied. The result showed that two rearrangements of TCR alpha gene detected with J alpha probes were the products of the normal TCR rearrangement process, and were not involved in the translocation at this region. This patient, together with the AST-1 cell line, provided us a unique opportunity to study the development and clonal evolution of malignant lymphoma.     

Phenotypic and Genotypic Lineage Switch of a Lymphoma with Shared Chromosome Translocation and T-Cell Receptor γ Gene Rearrangement

A case of non-Hodgkin's lymphoma showed a phenotypic and genotypic cell lineage switch twice during nine years of his clinical history; first, T-cell type, pleomorphic small cell lymphoma developed, followed by B-cell type, diffuse centroblastic/centrocytic lymphoma, and finally T-zone lymphoma without follicles again developed, from which AST-1 cultured cell line was established. Karyotype analysis demonstrated a shared abnormal chromosome, der(1)t(1;?)(p36;?), among the first relapsed B-cell tumor, the second relapsed T-cell tumor and AST-1 cell line. Furthermore, T-cell receptor (TCR) γ gene rearrangement bands of the same size were observed in the first relapsed B-cell tumor and the second relapsed T-cell tumor as well as AST-1 cell line. These results suggested that both relapsed tumors of different cell lineages are derived from a common malignant clone, presumably a committed lymphoid stem cell. A unique translocation, t(2;14)(q37;q11.2), which may involve TCR δ/α gene complex, was observed in the second relapsed tumor and AST-1 cells. To attempt to isolate the breakpoint of this translocation, the configuration of TCR δ/α gene complex was studied. The result showed that two rearrangements of TCR α gene detected with J_α probes were the products of the normal TCR rearrangement process, and were not involved in the translocation at this region. This patient, together with the AST-1 cell line, provided us a unique opportunity to study the development and clonal evolution of malignant lymphoma.