Surface markers and functional properties of non-Hodgkin's lymphoma cells in relation to histology.

Cells from 32 adult patients with non-Hodgkin's lymphoma were studied with respect to surface markers and functional properties in short-term culture. Twenty-six lymphomas were of B-cell origin, including all nodular and diffuse lymphocytic lymphomas. Three tumors were of T-cell origin (one histiocytic lymphoma and two undifferentiated lymphomas). In the remaining three cases (histiocytic lymphomas) the immunological nature of the tumor cells could not be determined. All reactivity to mitogenic stimuli of cells from B-cell lymphomas was due to residual normal T cells. In follicular lymphocytic lymphomas more reactive T cells prevailed among the malignant B cells than in diffuse lymphocytic lymphomas. Heterogeneity among B-cell lymphomas was indicated by differences in intensity of fluorescence with anti-Ig reagents and in stimulatory capacity in mixed lymphocyte culture. T-cell lymphomas were characterized by high percentages of T cells together with impaired responses to stimuli. The results of immunological studies correlated well with the histological classifications of Rappaport, Lukes and Lennert.     

Surface phenotyping, histology and the nature of non-Hodgkin lymphoma in 157 patients.

In a study of 157 patients with lymphoid malignancy, the phenotype of the tumour cells was correlated with the histological classification of the tumour using the Rappaport and the Kiel classifications. The markers used included E, Fc gamma, Fc micron (IgM) and C3d rosetting, estimation of SIg and CyIg, and tests for the expression of HTLA, Ia and ALL. Repeat biopsy specimens were studied in 23 of these patients. The phenotypic features of lymphoblastic malignancy indicated B-cell, T-cell and ALL-positive null-cell tumours in this group. Immunoblastic lymphomas were predominantly of non-capping B-cell type, but T-cell immunoblastic lymphoma occurred in 2 patients. Immunoblastic lymphomas of receptor-silent cells occur, and are ALL- and HTLA-negative. In the category of diffuse, poorly differentiated lymphocytic lymphomas, most cases are of centroblastic and centrocytic tumour of diffuse type, but pure centrocytic tumours and centroblastic tumours occur. The dominant phenotype in this group is of B cells expressing C3d receptors. Nodular poorly differentiated lymphocytic lymphomas (Rappaport) are classified as centroblastic and centrocytic follicular (Kiel) and most express SIg+ C3d+ phenotype. The frequency of this phenotype appeared the same in both diffuse and nodular poorly differentiated lymphocytic neoplasms. The Rappaport group of diffuse well-differentiated lymphocytic lymphoma includes 2 Kiel categories, malignant lymphoma lymphocytic, and malignant lymphoma lymphoplasmacytoid. Cells of the former tumour were considered to be immature B cells resembling those seen in CLL, and characteristically expressing SIg weakly, with a high frequency of single kappa light chain. Cells of the latter tumour are by contrast mature, and are related to the centroblastic and centrocytic follicular tumour by their histogenesis and phenotypic features. Repeat biopsy examinations indicate that T-cell predominance occurs in the prodromal phase of B-cell-predominant tumours of SIg+ C3d+ phenotype. It is concluded that non-Hodgkin lymphoma can be divided into 2 categories: (1) tumours of immature immunologically incompetent cells of lymphoblastic histology and with phenotypic features akin to T, B and Null-cell ALL, and (2) tumours of differentiated lymphocytes expressing the phenotypic features of B lymphocytes, with maturation arrested at one of several stages of an antigen-dependent immune response.     

Correlation of prognostic factors and blood lymphocyte subtypes in non-Hodgkin's lymphoma

The total lymphocyte, T, B, and null cell content of peripheral blood from 32 healthy individuals and 30 patients with non-Hodgkin's lymphoma was determined. The patients had a significant reduction of B cells (complement receptor and membrane immunoglobulin positive cells) and a significant reduction in T cells. Correlation of patients' characteristics with lymphocyte abnormalities demonstrated several findings. Patients with advanced disease (III and IV) had significantly lower total lymphocyte and T cells than patients with localized disease (I and II) or normal controls. Patients with hypogammaglobulinemia had lower total lymphocyte and T cells than patients with normal gamma globulin status or normal controls. Patients with diffuse histology and B symptoms had lower total lymphocyte and T cells than normal controls. Discriminant analysis of lymphocyte populations categorized patients by disease extent and gamma globulin status with 70 and 68% accuracy, respectively.     

Terminal deoxynucleotidyl transferase activity in lymphoma.

Terminal deoxynucleotidyl transferase (TdT) was estimated in the tissues of 42 patients with lymphoma, whose cells were also typed by the use of surface markers. Four of the 8 patients with T-cell lymphoma were TdT+ including patients whose lymph nodes showed an undifferentiated or poorly differentiated appearance. The TdT- T-cell lymphomas included cases with diffuse histiocytic Sezary cell, diffuse, poorly differentiated and angio-immunoblastic histology. The tissues of 31 patients with B-cell lymphoma were invariably TdT-, whether the histology was poorly differentiated, well differentiated, nodular, diffuse, histiocytic or Burkitt type, and including cases with about equal proportions of T and B cells, and those whose cells showed non-capping and capping surface immunoglobulin. Hodgkin's tissue was also invariably TdT-. We conclude that estimation of TdT in tissues of patients with malignant lymphoma may be a useful test in diagnosing the T-cell lymphoma, particularly in patients with tumours of undifferentiated or poorly differentiated histology.     

Peripheral blood lymphocyte subsets in nasopharyngeal carcinoma

Peripheral blood lymphocyte subsets were studied in 29 untreated Chinese nasopharyngeal carcinoma (NPC) patients and 20 normal controls of similar age range and sex ratio with indirect immunofluorescence using monoclonal antibodies. The percentage T lymphocyte (T11) count, percentage and absolute T helper (T4) lymphocyte counts and the T helper/T suppressor-cytotoxic cell ratio (T4/T8) were significantly lower, while the percentage and absolute T8 counts were significantly higher in NPC patients. Concerning B lymphocytes, B1 cell count was normal while B4 cell count (B lymphocytes including early B lymphocytes) was significantly lower in NPC patients. These various lymphocyte subset changes were not related to the clinical stage or to Epstein-Barr viral IgA viral capsid antibodies and IgA early antibodies. The pathogenetic and prognostic significance of peripheral blood lymphocyte changes in NPC requires further investigation.     

CCNU in combination chemotherapy for advanced histologically unfavorable non-Hodgkin's lymphoma

During a 3-year period 39 evaluable patients with stage III and IV non-Hodgkin's lymphomas and unfavorable histologies were treated with a unique chemotherapeutic regimen based on a modified CHOP combination to which was added the nitrosourea, CCNU. Complete response was observed in six of 15 (40%) patients with diffuse poorly differentiated lymphocytic lymphoma (DPDL), four of 11 (36%) with diffuse mixed histiocytic lymphocytic (DML), and seven of 13 (54%) with diffuse histiocytic lymphoma (DHL). Of the 17 patients who achieved complete response, nine (53%) have remained continuously disease-free for greater than 2.5 years (2.7-4.1 years) from the onset of therapy: four of six with DPDL, two of four with DML, and three of seven with DHL. Median survival was 18.9 months for all patients, 18.9 months for those with DPDL, 17.4 months for those with DML, and 9.7 months for those with DHL. The median survival has not been reached for patients who attained a complete response, and will exceed 3.3 years. Central nervous system relapse was observed in three patients. In general, toxicity was moderate and consisted primarily of leukopenia, nausea, vomiting, and neurotoxicity. There were no drug-related deaths. The addition of CCNU to a modified CHOP combination resulted in an effective, generally well-tolerated out-patient regimen. However, it did not appear to decrease the rate of CNS relapse or improve current treatment results observed with other adriamycin-containing regimens for similar patients.     

T and B cell populations in blood and lymph node in lymphoproliferative disease.

Lymph node and peripheral blood lymphocytes were studied simultaneously for surface markers of T and B cells in 22 patients with lymphoproliferative diseases and 8 patients with non-neoplastic lymphadenopathy. This resulted in the classification of the malignancy from involved lymph nodes into 4 groups. Six patients had B cell lymphomata with normal or strong immunofluorescent staining for surface membrane immunoglobulin; 8 patients had B cell chronic lymphocytic leukaemia with pale staining for surface membrane immunoglobulin; 5 patients had T cell lymphomata and 3 patients were not definitely classifiable. In 6 out of 8 patients with B cell CLL, histopathology of lymph nodes showed infiltration with well differentiated lymphocytes and in all T cell lymphomata, the infiltrating cells were poorly differentiated. By the use of these markers, malignant lymphocytes were identified in the circulation in only 3 out of 6 patients with B cell lymphoma, in all patients with B cell CLL but in none of those with T cell lymphoma or unclassifiable lymphoma. Therefore a more conclusive characterization of the malignant lymphocyte in lymphoproliferative diseases must include an examination of involved lymph nodes.     

表面增强激光解吸/电离-飞行时间质谱技术检测不同来源淋巴瘤细胞株及健康人外周淋巴细胞的差异蛋白质

 目的　应用表面增强激光解吸/电离-飞行时间质谱（SELDI-TOF-MS）技术对体外培养的人类T、B淋巴瘤细胞株和健康人外周血淋巴细胞的蛋白质进行检测，寻找差异蛋白。方法　常规培养人类T、B淋巴瘤细胞株，并从健康人外周血中分离出淋巴细胞培养，取对数生长期的细胞进行试验，使用蛋白提取液提取蛋白质，应用SELDI-TOF-MS技术进行检测，采用SPSS 13.0统计软件包进行统计学分析。结果　T、B淋巴瘤细胞株与健康人外周淋巴细胞比较，均有6个差异蛋白质。T淋巴瘤细胞株与B淋巴瘤细胞株比较有7个差异蛋白质。结论　T、B淋巴瘤细胞间及二者与健康人外周淋巴细胞比较，蛋白质组学水平均有差异。应用SELDI-TOF-MS技术对筛选淋巴瘤的分子标志物有一定价值，并可能为研究淋巴瘤的发病机制和寻找治疗靶位打下初步基础。     

In vitro functional studies of mononuclear cells in patients with CLL: evidence for functionally normal T lymphocytes and monocytes and abnormal B lymphocytes.

In vitro functional studies of mononuclear cells from 34 patients with B-cell type CLL were investigated and the results of these studies were as follows: 1) The T lymphocytes from patients with CLL were capable of responding normally to PHA or PWM, of inducing allogeneic normal B lymphocytes to respond to these mitogens and of stimulating normally to allogeneic lymphocytes in "one-way" mixed lymphocyte reaction; 2) The monocytes from these patients were capable of enhancing the T lymphocyte response to mitogens and of stimulating normally to allogeneic lymphocytes; and 3) The leukemic B lymphocytes were incapable of responding to mitogens even in the presence of normal T lymphocytes and their enhancer cell activity on T lymphocyte response or their stimulating capacity on allogeneic lymphocytes was depressed. These observations suggest that the T lymphocytes and monocytes from patients with CLL are functionally normal while the leukemic B lymphocytes from these patients are functionally abnormal.     

恶性淋巴瘤患者外周血T淋巴细胞亚群变化的讨论

目的探讨恶性淋巴瘤患者T淋巴细胞亚群的变化特点。方法采用流式细胞术检测24例各型恶性淋巴瘤患者及20例正常人的外周血中T淋巴细胞(CD3+细胞)、辅助性T细胞(CD4+)、抑制性细胞毒T细胞(CD3+/CD8+)、NK细胞(CD3-/CD16+56)和NKT细胞(CD3+/CD16+56),对两者百分比进行比较分析。结果患者组中辅助性T细胞(CD3+/CD4+/CD8-)的百分比和绝对值明显低于正常对照组(P0.01),而患者组中CD4/CD8细胞亦明显低于正常对照组(P0.01)。结论在恶性淋巴瘤发生发展的过程中了解淋巴细胞亚群的变化,可为后期病情判断及可能采取的相关治疗提供依据。     

Immunophenotype of lymphocytes of patients with malignant melanoma and its dynamics during immunotherapy with interleukin-2]

A Mab set was used to study immunological phenotype of lymphocytes of five patients bearing stage IV malignant melanoma and its changes developing in the course of interleukin-2 (Euro cetus) immunotherapy. Patients were shown to have lowered counts of T cells (mainly due to T-helper/inducers), B cells and lymphocytes expressing adhesion and activation molecules as well as abnormal immunoregulatory cell ratio. Two patients receiving interleukin-2 achieved a response. Follow-up of antigen expression revealed a rise in natural killer and immunoregulatory T lymphocyte levels in the responders whereas activation antigens and B cell levels were increased in all the patients.     

iNOS在弥漫大B细胞淋巴瘤组织中的表达及临床意义

目的 探讨诱导型一氧化氮合酶（inducible nitric oxide synthase,iNOS）在弥漫大B细胞淋巴瘤（diffuse large B cell lymphoma,DLBCL）组织中的表达及临床意义。方法 用免疫组化法检测40例DLBCL及15例反应性增生淋巴结中iNOS的表达情况,分析iNOS的表达与DLBCL患者性别、年龄、临床分期、国际预后指数（IPI）之间的关系。结果 iNOS在DLBCL及反应性增生淋巴结组织中的阳性表达率分别为67.5%、26.7%,两者比较差异有统计学意义（P<0.05）。iNOS的表达水平与临床分期、IPI有关（P<0.05）,与患者性别、年龄无关（P>0.05）。结论 iNOS在DLBCL组织中有较高的阳性表达,其表达水平与患者的临床分期及IPI相关,提示iNOS在DLBCL的发生、发展过程中起重要作用。     

Lymphocyte markers in non-Hodgkin's lymphomas.

The lymphocyte marker pattern of non-Hodgkin's lymphoma cells was related to current concepts of lymphoma classification. In a series of 28 lymphomas lymphocyte markers indicated that 2 were of histiocytic origin, 2 were unclassifiable, none were derived from T cells and the remainder were B-cell neoplasms. The immunoglobulin heavy chain associated with the B-cell tumours was gamma in one case, alpha in one case but was mu in the majority of cases, reflecting the predominance of this heavy chain, together with delta chains, on normal lymph node lymphocytes in man. delta chains accompanied mu chains on the tumour cells in 6/17 lymphomas in which anti-delta staining was performed. delta chains were not found on any lymphomas other than well differentiated diffuse lymphocytic types. There was evidence of a reduction in surface immunoglobulin, Fcgamma and C3 receptors on undifferentiated lymphoma cells. T lymphocytes of normal morphology were present in all lymphomas except one, and were more numerous in follicular lymphomas than in diffuse tumours.     

Colony formation by normal and malignant human B-lymphocytes.

A method is described that permits colony formation in culture by B lymphocytes from normal blood and from blood, marrow or lymph nodes of patients with myeloma or lymphoma. The method depends on: (1) exhaustively depleting cell suspensions of T lymphocytes, (2) a medium conditioned by T lymphocytes in the presence of phytohaemagglutinin (PHA-TCM), and (3) irradiated autologous or homologous T lymphocytes. Under these conditions the assay is linear. Cellular development of B lymphocytes can be followed; differentiation to plasma cells is seen in cultures of cells from normal individuals and myeloma patients, but not lymphoma patients. Malignant B lymphocytes in culture produced immunoglobulin of the class identified in the patient''s blood, or in freshly obtained cells. We conclude that the assay is suitable for studying the growth, differentiation and regulation of normal and malignant B lymphocytes in culture.     

The significance of bone marrow involvement in non-Hodgkin's lymphoma: the Eastern Cooperative Oncology Group experience

Data from four clinical trials conducted by the Eastern Cooperative Oncology Group (ECOG) were used to investigate the importance of bone marrow involvement as a prognostic factor in patients with non-Hodgkin's lymphoma (NHL). A total of 502 patients, 275 with nodular, poorly differentiated lymphocytic lymphoma (NLPD) and 227 with diffuse histiocytic lymphoma (DHL) or diffuse mixed-cell lymphoma (DML), were included in this analysis. Patients were separated into four categories: stage III, stage IV with bone marrow involvement (stage IV-M), stage IV without marrow involvement (stage IV-O), and stage IV with bone marrow and other organ involvement (stage IV-OM). Among the DHL and DML patients, the incidence of marrow involvement was 23%. However, stage IV-M patients had a prognosis that is similar to stage IV-O and stage IV-OM and worse than stage III patients. In contrast, the incidence of involvement with NLPD was 59% and patients with stage IV-M had a survival not different than stage III and not worse than stage IV-O and stage IV-OM. The results suggest that the current emphasis on bone marrow biopsy(s) as a routine diagnostic staging procedure for patients with NHL should be reevaluated. The necessity for this procedure in stage III patients with NLPD is not apparent from our data. One can still justify a bone marrow biopsy in stage I and II patients and can confirm the complete clinical response when all nodes have regressed in more advanced disease.     

Neutral glycosphingolipid expression in B-cell neoplasms

The expression of neutral glycosphingolipids (GSL) in 37 B-cell neoplasms [7 acute lymphocytic leukemia (ALL), 5 Burkitt's lymphoma (BL), 7 chronic lymphocytic leukemia (CLL), 5 diffuse, poorly differentiated lymphoma (DPDL), 6 diffuse histiocytic lymphoma (DHL), 3 hairy-cell leukemia (HCL), and 4 multiple myeloma (MM)] was examined. Patterns of expression of simple (GlcCer, LacCer) and globo-series GSL (Gb3, Gb4) were found for each tumor type. In addition, pre-B ALL expressed the neo-lacto series GSL, paragloboside, which was not significantly seen at later stages of maturation. As a group, leukemias expressed about 10 times higher ratios of simple GSL to Globo-series GSL as compared to lymphomas, regardless of stage of differentiation. Significant amounts of GSL of other series were not found except in one CLL which contained asialo-GM2. GSL phenotype in these cells was not grossly affected by cell genotype since pre-B ALL containing Philadelphia chromosome t(9q;22q) translocations were similar to other ALL; and DHL with t(8q;14q) translocations had GSL patterns similar to other DHL samples and dissimilar to GSL patterns found in Burkitt's lymphomas with t(8q;14q). Differences in GSL expression among the different types of B-cell neoplasm suggested that GSL patterns form a phenotypic map that may complement the traditional glycoprotein immunophenotypic map and contribute to our understanding of the biology of these diseases and B-cell differentiation.     

Nodular Lymphocyte Predominant Hodgkin Lymphoma

Nodular lymphocyte predominant Hodgkin lymphoma (NLPHL) is a rare subtype of HL with unique clinicopathologic features. The hallmark histologic feature is the presence of malignant LP cells, unusual CD20⁺CD15^?CD30^? variants of Reed‐Sternberg cells, embedded within a nodular pattern of infiltrating lymphocytes. Compared with classical HL, NLPHL shows a slightly older median age at presentation (30–40 years), greater male predominance (3:1), less mediastinal involvement (<15%), and lower occurrence of classical HL risk factors. The differential diagnosis includes progressive transformation of germinal centers, lymphocyte‐rich classical HL, and T‐cell/histiocyte‐rich large B‐cell lymphoma, the latter of which may share a common biologic relationship. The vast majority of patients present with limited stage disease (70%–80%), the standard treatment for which is involved field radiotherapy at 30–36 Gy. Response rates to primary therapy exceed 90%, although relapses are common and may occur years after the initial diagnosis. Secondary malignancies, particularly non‐Hodgkin lymphoma, may also occur at a frequency similar to that of relapsed NLPHL. Patients with advanced stage disease may have lower response rates and overall survival times than those with limited stage disease. For relapsed disease, treatment options include the salvage therapies used in classical HL, and rituximab.     

The localization of Hodgkin's disease in lymph nodes. A study with immunohistological, enzyme histochemical and rosetting techniques on frozen sections.

Lymphoid tissue of 51 patients with Hodgkin's disease was studied with immunohistological, enzyme histochemical and rosetting techniques for the detection of B and T cells in frozen sections. In lymph nodes of patients with lymphocyte predominance type of Hodgkin's disease, the majority of the lymphocytes in the involved areas were normal B lymphocytes of polyclonal origin. This was also true for nodular sclerosis cases with a predominance of lymphocytes. Surrounding Sternberg-Reed cells small clusters of T lymphocytes could be demonstrated. In mixed cellularity and also in nodular sclerosis with a mixed cellular pattern only small residual areas of B lymphocytes were present, whereas relatively large numbers of T lymphocytes were found in the involved areas. In lymphocyte depletion B lymphocytes were scarce and T lymphocytes were present in small number. It is concluded that different patterns of lymphocyte population can be discerned in the subtypes of Hodgkin's disease. A predominance of B lymphocytes is found in cases with lymphocyte predominance and thus is a prognostic favourable sign. A predominance of T lymphocytes as found in cases with a mixed cellular pattern with or without nodular sclerosis is therefore not a favourable sign in general but may indicate progressive disease. The possible reasons for the presence of large numbers of B or T lymphocytes in lymphoid tissue affected by Hodgkin's disease are discussed.     

增生性Kikuchi淋巴结炎与恶性淋巴瘤的鉴别诊断

目的：探讨增生性Ｋｉｋｕｃｈｉ淋巴结炎与恶性淋巴瘤的鉴别诊断要点。方法：收集７例诊断疑难的Ｋｉｋｕ－ｃｈｉ淋巴结炎病例进行组织学和免疫组织化学观察。结果：本病的共特点是：（１）先发热后淋巴结肿大、有自限性;（２）病变为多灶性。由以新生组织细胞和吞噬细胞为主混有少量浆细胞样单核细胞、Ｔ免疫母细胞及Ｔ小淋巴细胞多细胞组成,有核碎片散在;（３）缺乏中性白细胞浸润;（４）核分裂易见。结论：作者认为本病的特殊病史,病变多灶性,以组织细胞为主的多细胞组成并有核碎片散在等特点是与恶性淋巴瘤朱鉴别诊断要点。     

Expression of leucocyte-common antigen and large sialoglycoprotein on leukemic cells in B-cell chronic lymphocytic leukemia and non-Hodgkin's lymphoma

Patterns of leucocyte-common antigen (L-CA) and large sialoglycoprotein (LSGP) expression on leukemic peripheral blood lymphocytes of 13 patients with chronic lymphocytic leukemia (CLL), 17 with non-Hodgkin's lymphoma (NHL) in leukemic phase and one with hairy cell leukemia (HCL) have been examined by means of surface labelling and electrophoresis in 5% polyacrylamide gels. The 13 CLL, 10 of the 11 diffuse NHL and the six nodular poorly differentiated lymphocytic lymphoma (PDLL) patients fell into three groups according to expression of 210, 198 and 185k forms of L-CA. Group 1 (210 less than 198 less than 185k L-CA) included eight CLL and one diffuse NHL; Group 2 (210 greater than or equal to 198 and 185k L-CA) included four CLL, three diffuse NHL and four nodular PDLL; Group 3 (mainly 210k L-CA) included one CLL, six diffuse NHL and two nodular PDLL. A patient with diffuse large cell lymphoma and the HCL patient both had patterns of multiple, diffuse, very high Mr labelled glycoproteins. LSGP on these cells varied from nil to very high and levels were not related to L-CA patterns. Lymph node cells from five patients were also studied and were found to express larger numbers of L-CA forms and less LSGP than corresponding peripheral blood lymphocytes. Possible relationships of L-CA forms and LSGP to lymphocyte function and disease patterns are discussed.