Human T cell leukemia antigens on the cell membranes: purification molecular characterization, and preparation of specific antisera.
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Biomedical subjects
Publications and source records attributed to J Minowada.
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Adenosine deaminase (ADA) has been detected immunohistochemically in human thymus. The enzyme was localized predominantly in cortical thymocytes. Occasional lymphocytes in the medulla were also positive for ADA. Blood vessels, connective tissue, and Hassall's corpuscles were not stained for the enzyme. Using single-cell immunofluorescence and immunoperoxidase assays, we found that thymocytes and lymphoid cells of peripheral blood (PBL) and tonsils were heterogeneous with respect to ADA expression. About 70% of thymocytes were strongly stained for the enzyme whereas weak staining was seen in 20% of cells. About 10% of thymocytes were ADA negative. Twenty percent of PBL and tonsil cells were strongly positive for ADA, 10% of cells were negative for the enzyme, and weak staining was seen in the remainder. Bone marrow mononuclear cells were not stained for ADA. One hundred percent of lymphoblasts of 3 T cells leukemia lines were strongly stained for the enzyme whereas weak staining was seen in a pre-B cell leukemia line, 4 B cell lymphoma/normal lines, 2 non-T, non-B cell leukemia lines and 2 myeloid cell leukemia lines. There was a good correlation between intensity of cellular staining and quantity and activity of ADA detected in cell extracts by radioimmunoassay and enzymatically. The development of immunomorphologic methods for the detection of ADA provides a tool to study the role of the enzyme in function(s) and differentiation of normal and leukemic cells.
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A seasonal variation was observed when aryl hydrocarbon hydroxylase (AHH) activity was measured in the cultured lymphocytes of 977 donors over a period of 2 1/2 years. The variation was strongest in AHH activity induced by 3-methylcholanthrene and was less apparent for AHH activity in lymphocytes grown without any inducer. The period of the seasonal variation is 1 year, and maximal induced AHH activity occurs during late summer and early fall with minimal activity 6 months later. Based on the average of all individuals tested during the highest and lowest weeks, induced AHH activity can be as much as 10-fold higher during the peak season. It is not possible to tell from these experiments whether the seasonal variation is tissue specific, occurring only in lymphocytes, or characteristic of microsomal oxidases in other tissues as well.
We report here the identification of human cell-surface glycoprotein with unusual and interesting properties. We initially detected this glycoprotein on the surface of cultured human haematopoietic cell lines by means of a monoclonal antibody. Although it is expressed on all cultured human haematopoietic cell lines tested, including the inducible promyelocytic tumour cell line HL-60 (ref. 1), it is not present in readily detectable amounts on most normal or leukaemic human haematopoietic cells. HL-60 cells, on exposure in vitro to appropriate chemical inducers, undergo morphological and functional differentiation along the granulocytic pathway or into macrophage-like cells. One consequence of in vitro induction in the specific loss of the glycoprotein from the surface of HL-60 cells. The molecule does not, however, seem to be a highly tissue-specific differentiation antigen because it is also found on human tumour cell lines derived from non-haematopoietic tissues. Rather, its expression seems to be related to cell proliferation. Preliminary chemical characterization suggests that the glycoprotein maybe identical to the abnormal glycoprotein previously reported by Bramwell and Harris to be associated with malignancy.
We report the isolation and establishment in continuous culture of a human lymphoid cell line (Peer) from a case of T-leukemia. The Peer cell line lacks some typical cell-surface properties of T cells, namely sheep erythrocyte rosette formation and reactivity with two anti-T-cell sera, but has focal acid phosphatase and does express two other T-cell antigens, one defined by a monoclonal antibody, the other related to a T-cell subset (TH2). The cells are negative for B-cell markers (SmIg or cytoplasmic mu Fcgamma and C3 receptors, mouse erythrocyte rosettes) and EBV (EBNA). In addition, the Peer cell does not possess the typical phenotypic markers of "non-B, non-T" leukemia: cALL and Ia-like antigens, and the cytoplasmic hexosaminidase isoenzyme I, but is positive for terminal deoxynucleotidyl transferase by enzymatic and immunofluorescent criteria. The cell line requires exogenous L-asparagine for adequate growth in culture, a property known to be characteristic of certain T cells but not of B cells. The Peer cell line appears to have a maturation arrest at a developmental stage intermediate between the cortical thymocyte and a mature T-cell subset and to have lost some T-cell differentiation features.
Human hematopoietic cell lines, NALM-1, derived from Ph'-chromosome-positive chronic myelocytic leukemia (CML) in blastic crisis, and BALM-2, derived from B-cell acute lymphoblastic leukemia (ALL), were tested for their heterotransplantability and clonability. Two previously established cell lines, a Burkitt's lymphoma cell line, B35M, and a B-lymphoblastoid cell line, B411-4, derived from a hematologically normal person, were used for comparison. Ten million cells were needed in order for the BALM-2 cell line to grow with 74% success in the immunosuppressed mice, but 1000 million cells were needed in order for the NALM-1 cell line to grow with 74% success in those mice. As in the case of the BALM-2 line, 10 million cells of the B35M cell line were needed for 100% success in heterotransplantation, whereas 30 million cells of the B411-4 cell line resulted in 20% success of heterotransplantation. In a semisolid agarose (0.15%) medium, the cloning efficiencies of BALM-2, B35M, and B411-4 were 1.52--2.08, 26.31--38.46, and 0.09--0.12, respectively. With as many as 25,000 cells per well plated, NALM-1 did not produce any colonies in the semisolid agarose medium. Unlike most previous findings, those of the present study suggest that the heterotransplantability and clonability of cultured cells may not be invariably reliable characteristics of all proven human leukemia-lymphoma cell lines. The behavior of these leukemia cell lines representing their respective original leukemia clones is interpreted as reflecting the heterogeneity inherent in the biologic characteristics seen in human leukemias.
The splenic T and B cell distribution in 79 patients with untreated Hodgkin's disease was quite similar to that in 15 control patients with non-Hodgkin's lymphoma or carcinoma. The mean T lymphocyte percentage was slightly higher in involved spleens than in uninvolved spleens of patients with Hodgkin's disease. There was no significant difference in the T and B cell distribution between tumor area and tumor-free area of the same spleens of Hodgkin's disease patients. Splenic T and B cell distribution did not correlate well with the clinical features of Hodgkin's disease. The splenic T cell percentage was significantly lower than that of the peripheral blood T cell percentage (P less than 0.05), while the splenic B cell percentage was significantly higher than peripheral blood B cell percentage (P less than 0.01) in 13 patients with untreated Hodgkin's disease. The splenic T lymphocyte response to PHA was significantly higher than the peripheral blood T lymphocyte response (P less than 0.05), and the splenic B lymphocyte response to PHA, in the presence of irradiated autologous splenic T lymphocytes, was also significantly higher than the peripheral blood B lymphocyte response (P less than 0.05) in 8 and 6 patients with untreated Hodgkin's disease, respectively. Since the control splenic cells, utilized in this study were obtained not from patients with non-neoplastic disease, but from patients with neoplastic disease other than Hodgkin's disease, our data are not conclusive, but only suggestive of normal T and B cell distribution and function in uninvolved spleens of patients with untreated Hodgkin's disease.
The present report describes the first case of well-differentiated nodular lymphocytic lymphoma evolving into Lennert lymphoma of T-cell origin. A 58-year-old white female developed malignant lymphoma, well-differentiated, lymphocytic type, nodular, with focal bone marrow involvement (stage IV) in May 1975. She received 16 cycles of cyclophosphamide and prednisone combination chemotherapy which was completed in October 1976. A complete remission was achieved. In December 1976, she relapsed and was treated with cyclophosphamide, vincristine, bleomycin, and prednisone until May 1977. Lymphadenopathy decreased until August 1978, but then increased again. Biopsy of an axillary lymph node was interpreted as Lennert lymphoma. She received methotrexate, cyclophosphamide, vincristine, adriamycin, and prednisone beginning in September 1978. When last seen in November 1979, she was in partial remission. Lymphoid cells obtained from lymph node which was involved with Lennert lymphoma consisted of 93% standard E-rosettes and 83% gravity E-rosettes. Cytoplasmic immunoglobulin on frozen sections was negative, but acid phosphatase (ACP) and alpha-naphthyl acetate esterase reactions were strongly positive. These findings support a T-cell proliferation in Lennert lymphoma. A review of the literature reveals only four cases of Lennert lymphoma of T-cell origin.
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Materials with lymphotoxin activity produced by lectin stimulated primary cultures of human lymphocytes enriched for T cells or B cells, as well as material obtained from established tissue culture lines of human lymphoid cells were tested for cytotoxicity towards a large number of primary cultures and established lines of human and animal cells. Highly selective effects were found. The patterns of responses to the various lymphotoxin preparations indicate heterogeneity not only among the different cell cultures but also among the lymphotoxin preparations. In addition to the heterogeneity of the various cell cultures in their responses to the lymphotoxin preparations, a spectrum of susceptibilities was also noted among clonal derivatives of the same parental lines. In quantitative terms the responses of the most resistant and the most susceptible cell cultures spanned approximately a 200-fold range. These findings could have bearing upon the mechanism of tumor evasion of host antitumor immunity.
Thirteen human T cell leukemia-lymphoma cell lines were examined to determine whether or not they released a product into the media in which they were growing which could stimulate granulocyte-macrophage colony formation by human hemopoietic cells. None of the cell lines studied released colony stimulating activity.
The present study describes a comparative study of MLR-S of various stages of fresh and cultured normal or leukemic myeloid, monocytic and erythroid cells. "One-way" MLR was performed, using a slightly modified whole blood method. Fresh leukemic myeloblasts from patients with acute myelocytic leukemia or chronic myelocytic leukemia, in blastic crisis possess a strong MLR-S whereas fresh granulocytes from patients with chronic myelocytic leukemia or from healthy subjects possess no MLR-S. Cultured leukemic myeloblasts from KG-1 or ML-1 cell line possess a very strong or a moderate MLR-S, whereas cultured leukemic promyelocytes from HL-60 cell line possess little or no MLR-S. Fresh leukemic erythroblasts or cultured leukemic erythroblasts from K-562 cell line exert a strong MLR-S whereas fresh erythrocytes exert no MLR-S. Cultured monoblasts from HPL-SK-1 or Murray cell line possess a very strong MLR-S whereas fresh monocytes from healthy subjects or from a patient with chronic monocytic leukemia possess a moderate MLR-S. These observations clearly indicate that there is a good correlation between the MLR-S and the cell differentiation stage. Observations in the present study also support the hypothesis that the MLR-S is a differentiation antigen which is completely lost by the terminal stage of myeloid or erythroid cell maturation or partially lost by the terminal stage of monocytic cell maturation.
A cell line has been established from blood lymphoblasts of a female patient with acute lymphoblastic leukemia (ALL) shown to have near-haploid (27 chromosomes) cells in the bone marrow. The findings about the cell line were: 1) The frequency of near-haploid cells in culture decreased with time from 98.2% when the culture was started to 5.4% 15 months later. 2) Most of the other cells except the near-haploid ones were hyperdiploid, i.e., duplicates of the cells with near-haploid chromosome constitutions. 3) Chromosome evolution was seen in the near-haploid clones. The possible ancestor clone (clone A) had 27 chromosomes, one of each pair except no.10, 14, 18, and 21, which were disomic. A suggested evolution process is: clone A yields clone B (26 chromosomes: clone A, -no.10) yields clone C (27 chromosomes: clone B, +X) yields clone D (26 chromosomes: clone C, -no.21), clone E (28 chromosomes: clone C, +NO.20). Clone B and D, each with 26 chromosomes, appeared to contain the lowest number of chromosomes, appeared to contain the lowest number of chromosomes ever described for human somatic cell clones in vitro. 4) Changes in the constitutions of the hyperdiploid cell clones were preceded by evolution and changes in the near-haploid clones. 5) In near-haploid cells with 2 X-chromosomes, 1 exhibited late DNA replication; in hyperdiploid cells with 3-5 X-chromosomes, 2 were non-late DNA-replicating. 6) Fresh (uncultured) and cultured leukemia cells were antigenically typical non-T, non-B or common type ALL cells (positive for la-like and null-type ALL antigens and negative for surface membrane immunoglobulin).
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In six patients with Epstein-Barr virus (EBV) induced infectious mononucleosis (IM) and in two patients with IM-like syndrome, peripheral blood lymphocytes in the acute and convalescent phase were tested for human la-like antigens as well as other cell surface markers. The major population of T lymphocytes in the acute phase showed la-like antigens, as detected by indirect immunofluorescence with heteroantisera, and the number of la-like antigen-positive T lymphocytes decreased with convalescence. The crossabsorption study indicated that the amount of la-like antigen on the surface of IM T cells was less than that of Raji cells.
This study describes the establishment of three non-Burkitt B-lymphoma cell lines (BALM-3, BALM-4 and BALM-5) originating from the pleural effusion of a patient with a poorly differentiated diffuse lymphocytic lymphoma. The cells of BALM-3, -4 and -5 exhibited a number of properties which distinguish them from the usual B-cell type lymphoblastoid cell lines. Thus, they lacked the Epstein-Barr virus genome and had abnormal chromosome constitutions including a 14q+ marker. The presence of the identical surface immunoglobulin isotypes (gamma and chi chain determinants), and Ia-like B-cell-associated antigen in the cultured cells and in the "fresh" lymphoma cells in vivo was demonstrated. These findings strongly suggested that these cell lines have B-cell characteristics and were derived from the original tumor cell population. BALM-5 cells, however, showed somewhat different growth, cell surface marker profile and functional characteristics compared to those of BALM-3, and -4 cells. These variations suggest that the BALM-5 cells were probably at different stages of B-cell maturation than those of BALM-3 and -4, even though all three cell lines (established in three separate flasks) originated from the cells of the same pleural effusion of a lymphoma with monoclonal B-cell characteristics.
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