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We developed a novel broadband electron paramagnetic resonance (EPR) technique using a tunable continuous-wave (CW) terahertz(THz) source. A photoconductive antenna (PCA) generates frequency-tunable THz waves in a wide frequency region of 0.05-1.1 THz as a beat frequency of two laser beams with different wavelengths. We successfully observed EPR signals of some samples at room temperature in a transmission setup. Advantages of this technique are its high dynamic range of homodyne detection and continuously tunable THz generation/detection. Thus, high-resolution EPR spectroscopy as well as frequency-swept EPR experiment will become possible using this technique.

In this article, we demonstrate terahertz (THz) electron paramagnetic resonance (EPR) spectroscopy using a frequency-tunable photomixing source/detection system to investigate the electronic structure of solid-state samples in a microscopic manner. Fiber-coupled photoconductive antennae were used to generate and detect continuous THz waves obtained as the beat signals from two laser beams with different wavelengths. We will show some examples of EPR spectroscopy obtained by multi-frequency EPR measurements at room temperature. The technique reported here has several advantages, including high spectral resolution, wide frequency-range operation, high dynamic range, owing to the continuous-wave (cw) and tunable generation/detection of THz waves.

The findings of morphologically dysplastic features in haemopoietic cells in de novo acute myeloid leukaemia (AML) has been named AML with trilineage myelodysplasia (AML/TMDS). We analysed the clinical data, karyotypes, and treatment outcomes of 230 de novo AML patients treated with the Japan Adult Leukaemia Study Group AML-87 protocol. 40 (17%) patients had AML/TMDS. Platelet count was significantly higher (P = 0.006) and bone marrow blasts were fewer (P = 0.01) in the AML/TMDS group than in the AML without TMDS. Abnormal karyotype was shown in 12/30 patients (40%) analysed. The complete remission (CR) rate for AML/TMDS was significantly lower than AML without TMDS (63% v 81%) (P = 0.01). The overall survival curves showed that the 40 patients with TMDS had a significantly worse survival than the 190 without TMDS (P = 0.0005). AML/TMDS also showed significantly worse disease-free survival (DFS) (P = 0.0001). Multivariate analysis revealed that the absence of TMDS in AML was the most significant factor in obtaining CR (P = 0.01) and a significant factor in predicting longer DFS (P = 0.04). Our data suggest that AML/TMDS responds poorly to intensive chemotherapy. Further study is required to determine the best treatment strategy for AML/TMDS and the biological differences between AML/TMDS and other types of AML.

A total of 541 consecutive patients treated between 1975 and 1985, 449 with B-lymphoma and 92 with peripheral T-lymphoma, excluding adult T-cell leukemia/lymphoma (ATL), termed peripheral non-ATL T-lymphoma, were analysed. Clinicopathological features that were predominantly associated with B-lymphoma were low and intermediate grades (defined by the Working Formulation), diffuse large cell type, stage II disease, bulky disease, favourable extranodal sites that are defined as a group of primary tumors such as the eye, Waldeyer's ring, thyroid, and stomach, while other features predominantly associated with peripheral non-ATL T-lymphoma were high grade pathology, diffuse mixed-cell and immunoblastic type, systemic "B" symptoms, poor performance status (PS), generalized lymphadenopathy, involvement of organs such as liver, skin, and nose, leucocytosis, and a high levels of serum alkaline phosphatase. The survival curve for B-lymphoma was better (P0.01) than that for peripheral non-ATL T-lymphoma. Multivariate analysis revealed that the major prognostic factors were pathology, stage, and primary site for B-lymphoma, while stage, PS, and total protein levels were important for peripheral non-ATL T-lymphoma. These results indicate that B- and peripheral non-ATL T-lymphomas appear to have different biological characteristics.

A newly established Ph1-positive cell line, KOPM-28 was analyzed for immunological phenotype and the release of biological substance. The surface marker analysis revealed myelomonocytoid antigens, erythroid antigens and megakaryocyte-platelet antigens. Also, the cells could be induced for hemoglobin synthesis by the addition of hemin. The conditioned medium of this cell line showed the presence of interleukin 1 (IL-1) activity but not interleukin 2. Exogenous IL-1 and the conditioned medium stimulated the DNA synthesis of the cells themselves. Consequently, the yield substance works on the growth of original cells as an autocrine fashion, and might act on other normal cells as a paracrine factor.

Twenty-one patients with ATL were assessed. The predominant physical findings were lymph node and bone marrow involvement, skin involvement, hepatosplenomegaly and leukemic manifestations. The predominant histopathological findings in both skin and lymph node specimens were the diffuse medium-sized cell type and the diffuse mixed cell type. Some phenotypic discrepancy was found between the neoplastic cells in the peripheral blood, lymph nodes and skin of patients with ATL with respect to CD45RA and CD45RO, and CD7, CD29, CD25 and HLA-DR. That is, the predominant neoplastic cell phenotype was the helper T-cell, which was CD3+, CD4+, CD7+, CD25+, CD45RA+ and HLA-DR+, and CD29- and CD45RO- in peripheral blood and lymph nodes, and CD3+, CD4+, CD7+, CD29+, CD45RO+ and HLA-DR+, and CD45RA- in the skin. In other words, we have described the phenotypic heterogeneity of ATL cells and demonstrated the heterogeneity of CD45R isoform expression on ATL cells in different organs--the skin, peripheral blood and lymph nodes--of the same patient.

Prognostic factors in 47 patients with pleural lymphocytic lymphoma developing in chronic tuberculous pyothorax were evaluated using Cox's proportional hazards model. There were 41 men and six women, aged 44-80 (median 61) years. Approximately 70% of the patients had localized disease in Stages I and II, and 30% advanced disease in Stages III and IV. Histologically, 27 patients had the diffuse large, immunoblastic type and 12 had others. In the other seven patients, histological subtyping of the lymphocytic lymphoma was impossible because of degenerative or necrotic changes in the histologic specimens. A diagnosis of lymphocytic lymphoma of B-cell type was made in one case using combined cytologic and surface maker findings on a cell suspension. In addition, immunologic and immunohistochemical studies revealed another 40 cases to be proven B-cell lymphomas. Poor performance status and elevated levels of BUN and GPT were significantly associated with shortened survival in a Cox's proportional hazards model. A poor performance status and high levels of serum BUN and GPT suggested a marked deterioration in a patient's condition. When compared with previous literature describing prognostic factors in patients with B-cell lymphomas and with lymphocytic lymphomas with unfavorable histologies or associated with long-standing inflammations, the only common prognostic factors was performance status. The significance of primary site in predicting survival from lymphocytic lymphoma is discussed.

By using human T-cell growth factor (TCGF), 10 cell lines were established from tissue samples of 10 patients with adult T-cell leukemia (ATL). Three cell lines were adapted to growth in medium lacking TCGF. The surface markers of all cell lines were characteristic of inducer/helper T cells, i.e., OKT3+, OKT4+, OKT6-, OKT8-, OKIa1+, and human Lyt2+ and Lyt3+, except that one cell line was OKT3-. The expression of the viral antigen was examined during establishment of 8 of the 10 cell lines. The viral antigen was not expressed in leukemic cells before cultivation. In 5 lines, the viral antigen was detected by immunofluorescent staining after a short period of cultivation. However, 3 cell lines, ATL-6A, ATL-9Y, and ATL-1K did not express the viral antigen during short-term culture: the ATL-6A and ATL-9Y cell lines became positive for the viral antigen after 5 and 2 months of cultivation, respectively; the ATL-1K cell line remained antigen-negative throughout a culture period of 13 months. Southern blot hybridization assay showed that all of the cell lines, including the viral antigen-negative ATL-1K cell line, contained the viral genome. Thus, the retrovirus was associated with all 10 cell lines established from ATL patients, but there was a heterogeneity in the expression time of the retroviral antigen in leukemic cells maintained in vitro. Our findings suggested that the expression of the viral antigen was not required for maintenance of the leukemic state in vivo and for growth of leukemic cells in vitro.

We studied surface markers present in 56 cases of lymphoma of the skin by immunohistochemical staining, using the ABC (avidin-biotin-peroxidase complex) and PAP (peroxidase-antiperoxidase complex) methods. Of these cases, 49 were T-cell lymphoma and 7 were B-cell lymphoma. Ten of the 49 cases of T-cell lymphoma were adult T-cell leukemia/lymphoma (ATL). Twenty-five of 31 cases of T-cell lymphoma except ATL analyzed by the ABC method showed a helper/inducer phenotype (Leu2a−, Leu3a+), two cases showed a suppressor/cytotoxic phenotype (Leu2a+, Leu3a−), one case showed Leu2a+Leu3a+, one case showed an inducer phenotype (Leu2a−, Leu3a+, Leu9+), and one case showed OKT11+, Leu2a−, Leu3a−, Leu1−, Leu9+, CD25+, Leu10+, CD30+. One CD8+ lymphoma was Pagetoid reticulosis, and a CD4+, CD8+ lymphoma was lymphomatoid papulosis with erythematous plaque. Cutaneous T-cell lymphoma (CTCL), previously described by Edelson et al., is defined as a helper T-cell lymphoma with marked affinity for the skin. In our study, 5 cases of T-cell lymphoma of the skin were not CTCL as described by Edelson et al. These results show that T-cell lymphoma of the skin is heterogeneous in nature. In other words, CTCL is one type but represents a major proportion of T-cell lymphomas of the skin.

A monoclonal antibody, MRK20, in F(ab′)2 form [MRK20‐F(ab′)2], which reacts with 85‐kDa membrane protein in a doxorubicin (ADM)‐resistant subline (K562/ADM) of human myelogenous leukemia cell line, K562, was examined for reactivity with 41 cultured human leukemia and lymphoma cell lines. None of these cell lines had ever been exposed to any anticancer agent in vitro except K562/ ADM. The relative resistance index to various drugs was calculated by dividing the 50% growthinhibitory concentration (IC50) of the test cell line by IC50 of K562 (the negative control in the antibody experiment). MRK20‐F(ab′)2 reacted with seven cell lines, KYO‐1 derived from chronic myelogenous leukemia in blastic crisis (CMLbc), CMK from acute megakaryoblastic leukemia, HEL from erythroleukemia, P31/FUJ from acute monocytic leukemia, KOPM‐28 from CMLbc, PL‐21 from acute promyelocytic leukemia and K562/ADM, MRK20‐F(ab′)2 did not react with 34 other cell lines. All seven MRK20‐F(ab′)2‐positive cell lines had relative resistance index values of 2 or more to anthracyclines (ADM, pyrarubicin, daunorubicin), mitoxantrone, etoposide, bleomycin, and pepleomycin. There was no distinct correlation between the reactivity to MRK20‐F(ab′)2 and a higher relative resistance index than 2 to vinca alkaloids, actinomycin‐D, cisplatin, 4‐hydroperoxycyclophos‐phamide, nimustine hydrochloride, methotrexate or cytarabine. These results indicate that MRK20‐F(ab′)2 detects a novel multidrug resistance to anthracyclines, mitoxantrone, etoposide, bleomycin and pepleomycin in cultured human leukemia and lymphoma cells.

A study was made on and-ATLA antibody among general inhabitants, those with hematologic disease and those with hepatic disease in Nagano Prefecture.Among 2, 299 inhabitants (990 males and 1, 309 females), anti-ATLA antibody was found to be positive in two males (0.1 %)The number of patients with hematologic disease was 155. Of them, malignant lymphoma accounted for 50 (T: 24.3 %, B: 37.8 %), acute leukemia 32 and other hematologic malignancy 28. No ATL was found among them. It was revealed that 56 of them underwent transfusion ranging from 2 to 346 units of blood, averaging at 49.7 units. The number of person whose anti-ATLA antibody was positive was one only, who was diagnosed as AML and was transfused 170 units.Of 42 patients with hepatic disease, HB virus-related liver disease (hepatitis, cirrhosis, hepatocellular carcinoma) accounted for 10, non-B group HBAg (-) for 18, alcoholic liver dysfunction 5 and other hepatic disorders 9. Three out of these 42 patients were found to be positive for anti-ATLA antibody, each three patients belonged to the “non-B” group (3/18, 16.6 %).The total number of positivist was thus six. Of them, five had a history of transfusion, and all five positivists excluding the one with AML were suffering from hepatic disease with HBAg (-).From the above, it was considered that in Nagano Prefecture, those whose anti-ATLA antibody was positive were conspicuously small in number, but main cause of the positivity was attributable to the transfusion. Most of the positivists were suffered from non-B hepatic disease with a history of transfusion, and the number of positivist was significantly greater even when compared with patients with hematologic disease with frequent transfusion. Accordingly, it is suggested that some of the hepatic diseases with HBAg (-) might be associated with HTLV.

Immunoblastic lymphadenopathy (IBL)-like T-cell lymphoma is a distinct peripheral T-cell lymphoma, which closely resembles angioimmunoblastic lymphadenopathy with dysproteinemia (AILD) and/or IBL, but is characterized by focal or sheet-like proliferation of immunoblasts and pale cells of T-cell nature. In this report, 36 patients with IBL-like T-cell lymphoma were analyzed. The disease is clinically characterized by generalized lymph node swelling, hepatosplenomegaly, fever, skin rash, polyclonal hypergammaglobulinemia, marked male predominance, predilection for the elderly, and poor prognosis. There was no association with human T-cell leukemia virus type I or human immunodeficiency virus. IBL-like T-cell lymphoma may be divided into two categories (CD4+ type and CD8+ type) by surface marker analysis. It can also be divided into three categories on the basis of the histologic findings of distribution of morphologically recognizable tumor cells: nine cases of “inconspicuous type,” six cases of “patchy type,” and 21 cases of “diffuse type.” Two cases of “inconspicuous type” converted later to “diffuse type.” DNA hybridization analyses in the ten recent cases revealed that three of four “inconspicuous types” and five of six “diffuse types” showed clonal rearrangement of T-cell receptor-beta chain gene without rearrangement of immunoglobulin heavy chain gene, providing strong evidence for clonal proliferation of T cells.

1255 cases of leukemia-lymphoma were tested between 1972 and 1984 by multiple marker analysis. Routine leukemia phenotyping was performed using standard morphological and cytochemical techniques in combination with clinical and histo-pathological information; the main emphasis was put on immunological surface marker analysis using erythrocyte rosette assays, TdT and a large panel of poly- and monoclonal antibody tests. The 1255 cases were divided into these major types and subtypes: 349 cases of ALL and related immature T- and Burkitt-lymphomas (cALL, pre B-ALL, B-ALL and Burkitt-lymphomas, T-ALL and immature, mostly leukemic T-lymphomas, Null-ALL), 454 cases of mature T- and B-cell malignancies (T-CLL, mycosis fungoides, Sezary-syndrome, T-lymphomas, B-CLL, hairy cell leukemia, multiple myeloma, B-lymphomas), 263 cases of acute myeloid leukemias (AML, AMMoL/AMoL), 182 cases of chronic myeloid leukemias (CML in chronic phase, CMoL, CML in blast crisis), 6 cases of erythroleukemia and 1 case of megakaryoblastic leukemia. A simplified classification scheme which has been used in our laboratories is presented. Phenotyping is of diagnostic, prognostic and therapeutic relevance, most evidently for patients with ALL. Routine leukemia phenotyping should be performed with highly standardized techniques and reagents and by combining information from several fields in the multiple marker analysis. New areas of leukemia research might become very useful for the routine procedure of phenotyping.

A 49-year-old man developed adult T-cell leukemia (ATL) and acute myeloblastic leukemia (AML) at the same time. Using Southern blotting analysis, the leukemic cells of the ATL were found to contain the human T-cell leukemia virus type I (HTLV-I) proviral genome, whereas those of the AML did not, indicating the HTLV-I not to be associated with the AML oncogenesis. At the initial presentation, the serum anti-HTLV-I antibody was judged on screening by a routine particle-agglutination (PA) test and an indirect immunofluorescence assay (IF) to be negative. By Western blotting analysis, however, the serum was proved to be positive for anti-HTLV-I antibody. These results indicate that a routine PA-test and an IF may show false negative reactions on very rare occasions of low antibody titer. This is the first report of a coincidence of ATL with another type of leukemia.

Three hundred consecutive cases of malignant lymphomas and lymphoid leukemias in the National Cancer Center Hospital during the last five years were evaluated in terms of multiple morphologic and functional parameters. Immunologically these cases were composed of 42 cases of nonT-nonB-, 135 cases of T-, 95 cases of 8-, and 28 cases of possible (defective) B-cell lymphomas and leukemias. The high ratio of T-cell lymphomas (45%) is noteworthy and 63% of them were of peripheral T-cell origin. Some of the peripheral T-cell lymphomas were difficult to subtype by the classical Rappaport classification and these were dealt with as separate entities. The lower frequency of follicular lymphoma in Japan was reconfirmed (22 of 95 B-cell lymphomas) and half of the B-cell lymphomas were diffuse large cell type. Morphologic characteristics, surface and other functional markers of the neoplastic cells in each subtype were presented in detail and discussed in relation to the normal differentiation and maturation of lymphoid cells. Cancer 50:2372-2382, 1982. MMUNOLOGIC STUDIES using T-, and B-lymphocyte I markers have greatly contributed to the classification of lymphomas and leukemia^.'-^ Rapidly accumulating data suggest that the neoplastic cells preserve their surface markers and immunologic function in close relation to the differentiation and maturation of normal lymphoid cell^.'^^-'^-'^ Malignant lymphomas reported in the United States (US) and Europe are mostly B-cell lymphomas and only 10-20% are T-cell lymph~mas.~-~~' 3-15 T-c ell lymphoma derived from mature peripheral T-lymphocytes is very rare in the US,'6,17 whereas it is common in In our series, T-cell lymphoma/leukemia occupied 45% of all cases and 62% of them were of peripheral T-cell origin. In the southern region of Japan the frequency

We describe five patients with adult T-cell leukemia/lymphoma (ATL) with neither integration of human T-cell leukemia virus type I (HTLV-I) into their leukemia cells nor anti-HTLV-I antibody in their sera. These findings indicate that HTLV-I may not have been involved in leukemogenesis in these patients. The clinicohematological, cytopathological, and immunological features of HTLV-I-negative ATL were exactly the same as those of HTLV-I-associated ATL. Leukemia cells with pleomorphic nuclei, generalized lymphadenopathy, hepatosplenomegaly, skin lesions, hypercalcemia, and elevated lactate dehydrogenase levels, all of which are characteristic features of typical ATL, were also seen in these patients with HTLV-I-negative ATL. Leukemia cells expressed T3, T4, and pan-T-cell antigens in three cases, and T3 and pan-T-cell antigens in two. All five patients had lived in ATL-nonendemic areas. The finding of HTLV-I-negative ATL suggests that factor(s) other than HTLV-I infection may be involved in ATL leukemogenesis.

Chromosome aberrations and clinical features of three patients with adult T cell leukemia-lymphoma (ATL) not associated with human T cell leukemia virus type I (HTLV-I) are described. From their clinical features, two patients were diagnosed as acute type and one patient was diagnosed as chronic type, which later converted to acute crisis. Clonal and many chromosomal abnormalities were observed before therapy in the two acute type cases and at relapse in the chronic type case. Karyotype aberrations, including trisomy 3, trisomy 7, trisomy 21, del(6)(q21), del(10)(p13), 14q11 translocation, and loss of X chromosome, all of which are frequently found in HTLV-I associated ATL, were also seen in these cases of HTLV-I-negative ATL.

Five cases of adult T cell lymphoma with hyperglobulinemia were reported. Two patients died of extension of the disease despite intensive chemotherapy. Three others are still under treatment. Histologically, focal proliferation of monotonous pale cells and foci composed of polymorphic large, medium, and small cells were characteristic of all five cases. Infiltration by eosinophilic leukocytes and/or small histiocytic aggregates were also recognized. In addition, proliferation of branching vessels with hyaline deposits, perivascular proliferation of "immunoblasts" and plasma cells, absence of residual follicles, and depletion of small lymphocytes were the findings, which were consistent with immunoblastic lymphoadenopathy (IBL). Varying degrees of polyclonal hypergammaglobulinemia also corresponded with those of IBL. Rosette assay of the lymph node cells revealed a mixed population of E receptor-positive large, medium, and small cells with atypical nuclei. They also had T cell antigens on the surface and revealed no terminal deoxynucleotidyl transferase activity. These findings suggested their peripheral T cell origin. The cellular characteristics of these five cases are discussed in relation to other adult T cell lymphomas and to the helper function of neoplastic T cells.

The case is that of a 21-year-old female with mediastinal and subcutaneous, tumors composed of sarcomatous growth of poorly differentiated histiocytes defined by enzyme histochemical staining, ultrastructural observation and detection of surface markers. At autopsy neoplastic cells in the solid tumor became less cohesive and pleomorphic with erythrophagia, while features in the lymph node draining from the tumor was compatible with the criteria of malignant histiocytosis. The place where such a sarcomatous variant of malignant histiocytosis should be placed within other histiocytic tumors is discussed and the literature reviewed.

A human eosinophilic leukemia cell line, designated as EoL, was established from the peripheral blood of a patient with Philadelphia chromosome-negative eosinophilic leukemia (EL). The EoL cell line grows in single cell suspension with a doubling time of 48 hours for about one year. The reactivity of these cells was tested with a panel of monoclonal antibodies; they were found to express surface IA antigen, myeloid antigen (IF10, MY9) and membrane receptors for interleukin 2 (IL-2, Tac antigen). Under standard culture conditions, a small percentage of cells having more typical eosinophilic characteristics was present. These cells had cytoplasmic granules and were positive for Luxol-fast-blue and eosinophil peroxidase. Under culture conditions to induce the maturation of myeloid cells, such as alkaline medium or addition of dimethyl sulfoxide (DMSO), the frequency of cells with typical eosinophilic features increased to about 40%. In addition, cytogenetic studies showed that cultured cells and original leukemic blasts presented similar chromosome abnormalities. EoL seems to be a unique leukemic line committed to the eosinophilic lineage and can provide a useful in vitro model for the study of malignant eosinophilic properties.

Tumor cells suspensions prepared from 129 B- or non-T cell malignancies were investigated with a panel of 10 monoclonal antibodies and conventional surface marker techniques. Surface immunoglobulin (sIg) and B1 antigen proved to be the most useful markers for B-cell lineage. Six major subtypes of acute lymphoblastic leukemia (ALL) of non-T cell nature are now recognized by these immunological techniques, including null-ALL, Ia-ALL, lymphoid stem cell ALL, pre-pre-B ALL, pre-B ALL and B-ALL. In cases of chronic leukemias and lymphomas of non-T cell nature, 80% of the tumor was defined by sIg and 88% by B1 antigen as definitely of B-cell lineage. The clonal character was also defined in 68% of the tumor on the basis of the detection of predominant single light chain in sIg. Ia-like antigen was detected in almost all cases (96%). Leukemic cells from all cases of chronic lymphocytic leukemia (CLL), chronic lymphosarcoma cell leukemia (CLsCL) and hairy cell leukemia (HCL) reacted with OKIa1 and anti-B1, and leukemic cells from most of them with anti-pan T monoclonal antibody (10.2). In more than half of CLL and CLsCL, leukemic cells were reactive with J5, OKM1, 9.6 and OKT8, but not with OKT3, OKT4 and OKT6. HCL cells had almost the same reactivity with these monoclonal antibodies as CLL and CLsCL cells except that J5 remained unreactive. These results indicated that Japanese CLL, CLsCL and HCL were different from Western ones at least with respect to surface marker characteristics. In cases of lymphomas, heavy chains of sIg were expressed in polyclonal fashion, especially in follicular lymphoma and diffuse lymphomas of medium sized cell type and large cell type, indicating that lymphomas of these types may originate from follicular center cells of the heavy chain switching stage. Anti-T monoclonals were also reactive with lymphoma cells. In about half of follicular lymphomas and diffuse lymphomas of the medium sized cell type, lymphoma cells reacted with 10.2, and less frequently with 9.6, OKT4 and OKT3. On the other hand, only in one or two cases of diffuse lymphoma of the large cell type and of immunoblastic sarcoma (IBS), did tumor cells react with 10.2 and 9.6, but this was exceptional. In more than 25% of IBS, tumor cells also reacted with OKT8, but not with OKT4 and OKT3. These results indicated that anti-T monoclonals are no longer specific for T-cell lineage.(ABSTRACT TRUNCATED AT 400 WORDS)

Thirty cases of immunologically determined and histologically diagnosed immunoblastic lymphadenopathy (IBL), angioimmunoblastic lymphadenopathy (AILD), and IBL-like T-cell lymphoma were clinicopathologically reviewed. Clinical manifestations and laboratory findings did not reveal significant differences in these three groups. IBL, AILD, and IBL-like T-cell lymphoma showed a spectrum of histologic changes, in which proliferation of pale cells was a critical diagnostic point for the histologic malignancy. Immunostaining for their subsets revealed that 3 of 21 cases showed T4+ phenotype and the remaining 19 cases showed T8+ phenotype. Three of seven immunohistochemically determined T8+ cases simultaneously expressed Leu7+ phenotype. The latter cells were consistent with large granular lymphocytes in one case, but no clinicopathological differences from the other T8+ cases were present. IBL and AILD were considered to be T-cell malignancies, which show a spectrum of histologic features from T-cell dysplasia to peripheral T-cell lymphoma (IBL-like T-cell lymphoma). Despite intensive chemotherapy, prognosis was poor in T8+ cases of which half of the patients died within 1 year. T4+ cases showed better prognosis, but a higher incidence of synchronous second primary cancers was recognized.

Prognostic factors in 68 consecutive patients with myeloma treated at the National Cancer Center Hospital from 1962 to 1984 were analyzed. Median survival time from onset was 100 months for stage I, 72 months for stage II, and 26 months for stage III of the Durie and Salmon's clinical staging system. It was 55 months in patients with normal renal function and 18 months in those with abnormal renal function. All early deaths occurred in patients with stage III disease. Hemoglobin level, bone lytic lesions and presence of Bence Jones protein were also significant prognostic factors. On the other hand, heavy chain as well as light chain subtypes of monoclonal immunoglobulin (M-component) and M-component production rate did not influence the survival of myeloma patients. The analysis of chemotherapeutic responses and survival curves according to the chemotherapy used in this study (alkylating agent vs Vinca-alkaloid plus alkylating agent) did not disclose any significant difference between the two groups. The overall response rate was 67%. The survival time from the initial chemotherapy of responding patients was significantly longer than that of nonresponders.

Cell suspensions or frozen sections of lymph node biopsies from 32 patients with non-Hodgkin's lymphoma (NHL) were studied for sheep erythrocyte (E)-binding under three conditions (Estandard, EAET, Egravity), Fc and C receptors, immunoglobulin (Ig) heavy and light chain class and reactivity with heterologous antisera to T cells (T-LCL), HLA-D (Ia-like) and common acute lymphocytic leukemia (c-ALL) antigens. Selected B and T cell lymphomas were also tested for reactivity with the monoclonal antibodies OKT 3, OKT 4, OKT 6, OKT 8, OKT 11A, Leu-1, Leu-2a, Leu-3a, Leu-4 and Leu-7. There were 26 B and 6 T lymphomas. Most B lymphomas were mu+ (81%), kappa+ (77%) and 31% were mu+ delta+. One of the T lymphomas arose in a patient with antecedent follicular small-cleaved (B) cell lymphoma. The most accurate marker for characterizing the immunologic phenotype in NHL was the clonal excess of kappa+ or lambda+ cells. Neither Estandard, EAET, Egravity or T-LCL were consistently reliable as sole reagents in identifying T-cell lymphomas, their individual scores often being lower than those of monoclonal pan-T cell reagents. HLA-D (Ia-like) antigen was noted in 89% of B and 50% of T lymphomas. The corresponding values for c-ALL antigen were 12 and 33%, respectively. The comparative scores in T-lymphomas between OKT 4 and Leu-3a for "helper-inducer" (HE) cells and OKT 8 and Leu-2a for "suppressor-cytotoxic" (SU) cells were not uniformly consistent. Four T lymphomas had a mixed HE/SU cell phenotype, one was HE, and another SU. Anti-T reactivity was detected in the neoplastic follicles of six of seven follicular lymphomas. The percentage of anti-T reactive cells within positive neoplastic follicles was usually small (5-15%) and of the same order as that noted within reactive lymphoid follicles (5-30%). High numbers (50-100%) of cells from five small lymphocytic B, three diffuse small cleaved cell B and six T cell lymphomas were also positive with one or more anti-T reagents, suggesting the presence of cross-reactive antigens that make phenotyping of lymphomas with monoclonal antibodies problematic. Reactivity with the monoclonal antibody Leu-7 (HNK-1), a putative NK-specific reagent, was seen in one of five B and three of five T lymphomas.

The modulation of cell surface antigens in 2 myeloblastic cell lines (ML-1 and KG-1), a promyelocytic cell line (HL-60), and a monoblastic cell line (THP-1-0) by the presence of 12-O-tetradecanoyl-phorbol 13-acetate [(TPA) CAS: 16561-29-8] was investigated by indirect membrane immunofluorescence with the use of three monoclonal antibodies (MoAb) (OKM-1, 63D3, and MCS-2) reacting with myeloid-monocyte antigens (expressed by cells of both granulocyte and monocyte lineages) and two MoAb (1/12/13 and MCS-1) reacting with myeloid antigens (expressed by cells of the granulocyte lineage). Functionally mature macrophage properties, such as adherence, morphologic character, and phagocytosis, were induced by the presence of TPA in each of the cell lines except for adherence in the HL-60 cells. After 3 days in culture, the expression of the OKM-1-defined antigen was markedly augmented in all 4 cell lines. The expression of the 63D3-defined antigen was also markedly augmented in the ML-1, KG-1, and THP-1-0 cells, but it was not significantly altered in the HL-60 cells. The MCS-2-defined antigen was amplified in expression in the ML-1 and HL-60 cells, but it showed minimum decrease in the KG-1 and THP-1-0 cells. The MCS-1-defined antigen expression was suppressed in ML-1, HL-60, and THP-1-0 but was enhanced in KG-1. The suppressed expression of My-1 antigen (detected by the MoAb 1/12/13) was noted in all 4 cell lines. Thus in the ML-1 cells, expression of the myeloid-monocyte antigens was augmented, whereas myeloid antigen expression was inhibited in the presence of TPA, a result that parallels antigenic expression in terminal macrophage differentiation. The trend was true, except for the 4 cell line-antigen combinations (MCS-2-defined antigen and MCS-1-defined antigen in KG-1, 63D3-defined antigen in HL-60, and MCS-2-defined antigen in THP-1-0). The heterogeneous attitude of some antigens to TPA found in these cell lines may result from the fact that they represent different points in the myeloid-monocyte differentiation scheme.

Forty cultured human leukemia and lymphoma cell lines never exposed to anticancer agents in culture, apart from doxorubicin (ADM)-resistant K562/ADM, were examined for reactivity with a monoclonal antibody, MRK16 in F(ab')2 form [MRK16-F(ab')2], which recognizes P-glycoprotein (P-gp). The relative resistance index to various drugs was calculated by dividing the 50% growth inhibitory concentration (IC50) of the test cell line by IC50 of K562, which was the negative control in the antibody experiment. MRK16-F(ab')2 reacted with four cell lines, K562/ADM, KYO-1, HEL and CMK, which had relative resistance index values of 2 or more to vincristine (VCR), vindesine, vinblastine, ADM, daunorubicin, mitoxantrone (MIT), etoposide (VP-16) and actinomycin-D (ACT-D). The level of resistance to VCR and ADM in these cell lines decreased significantly in the presence of 10 microM verapamil in vitro. Significant expression of mRNA of P-gp gene was also detected in K562/ADM, KYO-1 and HEL. MRK16-F(ab')2 did not react with 36 other cell lines. Among them, three cell lines, PL-21, P31/FUJ and KOPM-28, had relative resistance index values of 2 or more to anthracyclines, MIT and VP-16, but not to vinca alkaloids or ACT-D. The level of ADM-resistance in these cell lines did not decrease significantly in the presence of 10 microM verapamil. Five cell lines, ATL-1K, HL-60, KMOE-2, ML-1 and U266, had relative resistance index values of 2 or more to some of the drugs, but not to the others, and 19 other cell lines did not. These results indicate that the reactivity of MRK16-F(ab')2 correlates with a relative resistance index of 2 or more to all these drugs in cultured human leukemia and lymphoma cell lines.

Fresh tumor cells from pleural effusion of a patient with Hodgkin's disease were analyzed cytogenetically, immunologically and enzymocytochemically. They were characterized by the presence of alpha-naphthyl butyrate esterase activity, Fc gamma-receptor, HLA-DR antigen and No. 9 antigen which has been shown to be present in Hodgkin's cells and granulocytes, and the absence of definite T-, B- and myeloid cell markers. The karyotype analysis of these tumor cells revealed chromosome instability, but the clonality was confirmed by the many common abnormalities such as -4, -6, -10, -12, -13, -14, +21, del(X) (q22,q26), del(7) (q32q36), and +der(19)t(19;?). In addition, there were more duplicated tetraploid clones than near-diploid clones. The karyotype of the near-diploid clone was interpreted as: 48, X, del(X) (q22q26), -4, -6, -10, -12, -13, -14, +20, +21, +der(4)t(4;?) (p16;?), del(7) (q32q36), +der(19)t(19;?) (p13;?), +mar1, +mar2, +mar3. The karyotype abnormalities characteristic of lymphomas or leukemias were not found. These results indicate that the tumor cells are not of lymphoid or myeloid lineage. Further studies are needed to determine the cellular origin of the tumor cells of Hodgkin's disease.

We evaluated the occurrence and the type of second malignancies among 74 patients with Hodgkin's disease (HD) and 407 patients with non-Hodgkin's lymphoma (NHL) who were treated at the National Cancer Center Hospital for more than one year. Fifteen patients developed a second malignancy. In 10 of these patients the second cancer was gastric cancer, but no cases of acute nonlymphocytic leukemia were encountered. The observed number of second cancers in females among the HD patients was significantly (p less than 0.005) greater than the expected incidence based upon the number of age-adjusted person-years both for all cancers and for stomach cancer. However, no significant differences between males and females in the NHL patients were found. Furthermore, no significant differences were seen in any of the groups between the observed and expected numbers of second malignancies according to the treatment.

In ten human malignant T-cell lines (HPB-ALL, HPB-MLT, CCRF-CEM, RPMI-8402, MOLT-4, JM, P12/Ichikawa, TALL-1, SKW-3, HUT-102), the expression of the antigen defined by OKT-4 or Leu-3A monoclonal antibody was remarkably decreased after a short incubation (1 hr) with 12-0-tetradecanoylphorbol-13-acetate (TPA) (1.6 nM). This phenomenon was also observed in all types of cells tested, including two lymphoblastoid cell lines with B-cell properties (RPMI-6410 and RPMI-8392), thymus cells from three children, peripheral blood lymphocytes from four normal individuals, and fresh neoplastic T cells from a case of T-cell acute lymphoblastic leukemia (ALL) and a case of T-cell lymphoma. The presence of puromycin (10 microM), cycloheximide (35 microM), or retinoic acid (1.0 microM) had no effect on the TPA-induced loss of OKT-4 defined antigen. Incubation at 4 degrees C or with lidocaine (4 mM) partially prevented this effect of TPA, and the presence of sodium azide (150 mM) did so totally. Recovery of antigen expression occurred 24 hours after cessation of TPA treatment. Comparative study using several phorbols other than TPA showed that this effect accorded with the tumor promotor activity of the compounds, but the phenomenon appears to be at the membrane level, not necessarily as a result of TPA-induced cellular differentiation.

Our previous study suggested a close relation between a preceding chronic tuberculous pyothorax and the development of non-Hodgkin's lymphoma (NHL) in the pleural cavity. To confirm this further, 37 cases were collected from Japanese hospitals, and their clinical and pathological findings summarized. The age at first admission for lymphoma of patients ranged from 46 to 81 (mean 63) years, the male to female ratio being 5.2:1. All patients were admitted after a 22-55 (mean 33) year history of pyothorax resulting from artificial pneumothorax for the treatment of pulmonary tuberculosis (29 cases) or tuberculous pleuritis (seven cases). The most common presenting symptom was chest pain. The main tumor mass, detected by chest roentgenogram and computed tomographic scans, was situated in the pleura (28 patients), the lung near the pleura (five patients) and the pleura and lung (four patients). The diagnosis of pleural NHL was made by biopsy for 31 of the patients and at autopsy for the other six. Histologically 30 (81%) of the 37 cases were of the diffuse large cell type, and of these the immunoblastic type was the most common (22 cases). Immunological and immunohistologic studies revealed a B-cell nature of the proliferating cells in all but one tumor. Thirty-two patients received chemotherapy and/or radiotherapy. Twenty-seven patients died between one and 144 (median eight) months of diagnosis. Autopsies carried out in 23 cases revealed the disease to have been localized to the thorax in 11 patients. These findings indicated that malignant B-cell lymphoma arose as a monoclonal growth from a pool of proliferating polyclonal B lymphocytes in tissues affected by the chronic tuberculous pyothorax.

A panel of monoclonal antibodies (BA-1, BA-2 and BA-3) were made against the pre-B acute lymphoblastic leukemia (ALL) cell line NALM-6. BA-3 binds to the 100,000 mol. wt common ALL antigen (gp100/CALLA), BA-2 binds to a 24,000 mol, wt (p24) cell surface structure and BA-1 binds to a currently undefined antigen. These antibodies were analysed for their reactivity with 67 human leukemia/lymphoma cell lines. The cell lines studied included T ALL, non-T ALL, surface immunoglobulin+ leukemias/lymphomas, and myeloid/monocytoid leukemias. The results indicate that all three antibodies react primarily (but not exclusively) with malignant cells early in B cell development. This pattern of reactivity was similar to previous results obtained with fresh, non-cultured malignant cells. BA-1, BA-2 and BA-3 are useful tools for analysing the developmental options of normal lymphoid cells and for "cleaning up" leukemia/lymphoma cells in selected cases of autologous bone marrow transplantation.

We investigated the relationship between the expression of surface antigens and the cell cycle phase in leukemic cells from cell lines and one patient using two-color flow cytometry, in order to determine the reason for the uneven expression of some markers which frequently leads to equivocal results as to leukemic phenotyping. As a result, it was demonstrated that monocyte-related differentiation markers, including I2, My4, Mo1 and Mo2, on monocytoid leukemic cells are preferentially expressed at the GO/G1 phase. Consequently, it is expected that the positivities for such markers vary with the proliferation status of the leukemic cells.

Eighty-two adult patients with advanced B-lymphoma, treated between 1981 and 1983 with VEPA (vincristine, cyclophosphamide, prednisolone and doxorubicin) or VEPA-M (VEPA plus methotrexate) in a prospective randomized fashion, were evaluated for pretreatment characteristics. The overall complete response (CR) and the 4-year survival rates were 74% and 45%, respectively. The relapse rate was 51%. Stage of disease only was negatively associated with the CR rate in a multivariate analysis. The primary extranodal tumor site other than upper gastrointestinal (GI) tract and high grade pathology were found to affect disease-free survival adversely in a Cox proportional hazards model. Poor performance status, advanced stage, primary extranodal tumor site other than upper GI tract, advanced age, high grade pathology and prior therapy by either surgery or radiation, were significantly associated with shortened survival in a Cox proportional hazards model. These results indicate advanced B-lymphoma in Japan to be generally similar to advanced non-Hodgkin's lymphoma in the West in terms of prognostic factor characteristics, but the importance of the primary site in predicting survival has not been reported in the West. Also, the lack of a survival plateau in patients with diffuse large cell lymphoma indicates more intensive chemotherapy regimens than VEPA or VEPA-M to be needed. It was also found that the significant prognostic factors in patients with advanced B-lymphoma were very different from those with T-lymphoma. The five factors: pathology, stage, primary site, age, prior therapy by surgery or radiation, for which the risk ratio was more than 2.3, were used to construct a model containing 23 categories of patients running an increasing risk of shortened survival; this divided patients into three groups. The CR and 4-year survival rates of low-, moderate- and high-risk groups were 90% and 74%, 74% and 58%, and 50% and 5%, respectively. The risk-grouping provides indications for determining optimal therapy for individual patients and the need for new therapeutic trials in patients at high risk.