T-lymphocyte cell lines derived from patients with acute lymphoblastic leukemia.

Several lymphoid cell lines with thymus-dependent lymphocyte (T-cell) characteristics have been established from 2 patients with acute lymphoblastic leukemia (ALL). These cell lines are considered to be leukemic T-cells on the basis of their abnormal chromosome constitution, growth in heterologous animals, ability to form rosettes with sheep red blood cells and the absence of immunoglobulin production, and destruction by antithymus cell sera. Sera from patients with leukemia did not react with these cultured cells but there was a strong reaction with infectious mononucleosis sera despite the fact that the cultured leukemia T-cells had no detectable Epstein-Barr virus (EBV) nor its genome.     

Establishment and characterization of cell lines from Gross virus-induced rat leukemia.

Cells from thymus and/or bone marrow tissues of either normal or Gross virus-infected W/Fu rats were cultivated individually or mixed. Four continuous cell lines were obtained: 1) individual cultivation of the cells from thymic lymphoma, 2) combined cultivation of cells from thymic lymphoma and normal bone marrow, 3) bone marrow of a leukemic rat and normal thymus, and 4) bone marrow of a leukemic rat supplemented with cell-free extracts of normal thymus homogenates. The morphological features were similar in these 4 cell lines. The cell correspond morphologically to lymphoblasts, grow rapidly in suspension, and are transplantable in newborn rats. The cells release abundant Type-C virus particles. The inoculation of cell-free culture fluid into newborn rats resulted, after 7-9 weeks, in the development of thymic lymphoma in high incidence.     

Establishment and characterization of cell lines from gross virus-induced rat leukemia

Summary Cells from thymus and/or bone marrow tissues of either normal or Gross virus-infected W/Fu rats were cultivated individually or mixed. Four continuous cell lines were obtained: 1) individual cultivation of the cells from thymic lymphoma, 2) combined cultivation of cells from thymic lymphoma and normal bone marrow, 3) bone marrow of a leukemic rat and normal thymus, and 4) bone marrow of a leukemic rat supplemented with cell-free extracts of normal thymus homogenates.The morphological features were similar in these 4 cell lines. The cells correspond morphologically to lymphoblasts, grow rapidly in suspension, and are transplantable in newborn rats. The cells release abundant Type-C virus particles. The inoculation of cell-free culture fluid into newborn rats resulted, after 7–9 weeks, in the development of thymic lymphoma in high incidence.This work was supported in part by a Grant-in-Aid for Cancer Research from the Ministry of Education, Science and Culture, Japan.     

Establishment and characterization of four new squamous cell carcinoma cell lines derived from oral tumors

Summary Four cell lines were established from squamous cell carcinomas (SCC) of the oral cavity. Cell lines AW 13516 and AW 8507 were derived from poorly differentiated SCC and epidermoid carcinoma of the tongue respectively. Cell line AW 10498 was derived from moderately differentiated SCC of the lower alveolus, and AW 9803 grew from a well-differentiated SCC of a retromolar trigone. The cultures showed typical epithelial cell morphology, numerous mitotic figures, occasional multinucleated giant cells, individual cell diskeratosis and nuclear and nucleolar abnormalities. The cell lines AW 13516 and AW 8507 were fast growers with a doubling time of 35.5 h and 31.9 h, respectively, which was independent of the initial seeding density. Cell lines AW 10498 (doubling time 52.2 h) and AW 9803 (doubling time 66 h) showed slower growth and had shorter doubling times at higher seeding densities. The presence of cytokeratins was detected in all the four cell lines by using polyclonal antikeratin antisera in indirect immunofluorescence and in Western blotting. None of the cell lines expressed major histocompatibility complex (MHC) class II antigens. MHC class I antigens were expressed by three cell lines but not by AW 9803. Flow cytometric analysis of DNA content and chromosomal studies suggested the presence of polyploidy and aneuploidy in all the four cell lines. Ultrastructural studies revealed typical epithelial cell features, such as the presence of desmosomes, tonofilaments and keratin bundles.Abbreviations SCC squamous cell carcinoma - PBS phosphatebuffered saline - MHC major histocompatibility complex     

Establishment and characterization of a new human leukemia cell line derived from M4E0

A new human leukemia cell line, designated as ME-1, was established from the peripheral blood leukemia cells of a patient with acute myelomonocytic leukemia with eosinophilia (M4E0). This cell line has the characteristic chromosome abnormality of M4E0, inv(16) (p13q22). When cultured in RPMI 1640 medium containing 10% fetal calf serum, ME-1 cells were monoblastoid, but with the addition of cytokines such as interleukin-3 (IL-3), granulocyte-macrophage colony-stimulating factor (GM-CSF), IL-4, or medium conditioned by phytohemagglutinin-stimulated human peripheral leukocytes (PHA-LCM), the cells exhibited differentiation to macrophage-like cells. PHA-LCM also promoted eosinophilic-lineage differentiation of this cell line, although IL-5 did not do so. To elucidate the mechanism of proliferation and differentiation of ME-1 cells, we studied the effect of a potent inhibitor of protein kinase C, 1-(5-isoquinolinyl-sulfonyl)-2- methylpiperazine (H-7), on colony formation of ME-1 cells. H-7 inhibited colony formation of ME-1 cells by IL-3 or GM-CSF dose dependently, but had little inhibitory effect on colony formation by IL- 4. These results indicate that the proliferation and differentiation of ME-1 cells by IL-3 or GM-CSF were related to the activation of protein kinase C, while those by IL-4 involved other regulatory systems. ME-1 cells should be useful for studying the pathogenesis of M4E0 and the mechanisms of proliferation and differentiation of leukemic and normal progenitors by cytokines.     

Pulmonary complications in patients with adult T-cell leukemia]

In order to investigate the character of pulmonary complications in patients with adult T-cell leukemia (ATL), a pathological and bacteriological study was performed in 92 autopsy cases with hematologic malignancies including 17 cases of ATL and 103 autopsy cases with solid malignancies from 1981 to 1990. Among 17 cases with ATL, pulmonary complications were seen in 16 cases (94.1%); pulmonary infection in 14 (82.3%), leukemic cell pulmonary infiltration in 9 (52.9%), pulmonary hemorrhage in 5 (29.4%), pulmonary alveolar calcinosis in 2 (11.8%), and idiopathic interstitial pneumonia in 2 (11.8%). The causative microorganisms were virus in 10; 9 of which were cytomegalovirus, followed by bacteria infection in 4 cases, mainly pseudomonas aeruginosa, and fungal infection in 3, mainly cryptococcus. pneumocystic carinii and mycobacterium tuberculosis were not detected. It is suggested that patients with ATL are severely compromised with chiefly cellular immunodeficiency, and administration of sulfamethoxazole-trimethoprim and isoniazid is very effective in prevention of pneumocystis carinii pneumonia and pulmonary tuberculosis.     

Establishment and characterization of differentiated, nontransformed hepatocyte cell lines derived from mice transgenic for transforming growth factor alpha.

Hepatocytes are extensively used in studies of gene regulation but cannot be maintained in long-term culture as replicating, differentiated cells while remaining nontumorigenic. We have derived two hepatocyte lines from livers of transgenic mice overexpressing transforming growth factor alpha, a potent hepatocyte mitogen, which overcome these limitations. The transgenic hepatocytes were maintained for > or = 2 months in serum-supplemented primary culture and gave rise to cell lines, of which two (AML12 and AML14) have been cultured for > 1.5 years (> 80 passages). Both lines have typical hepatocyte features such as peroxisomes and bile canalicular-like structures, do not grow in soft agar, and are nontumorigenic in nude mice. Like normal hepatocytes, AML cells express high levels of mRNA for serum (albumin, alpha 1-antitrypsin, and transferrin) and gap junction (connexins 26 and 32) proteins, secrete albumin, and contain solely isozyme 5 of lactate dehydrogenase. After extensive passaging, AML12 cells continue to strongly coexpress hepatocyte connexin mRNAs but do not display nonparenchymal cell markers. Although mRNA levels for some serum proteins progressively fall, high expression in late AML12 cultures may be regained by passage in serum-free medium. The AML14 line loses expression of both differentiated markers and transgene mRNA with extended passaging, and hepatocytic traits are only partially restored by passage in serum-free medium. These differentiated, nontumorigenic cell lines should serve as models in which to study hepatocyte growth and differentiation.     

Infectious complications in patients with adult T-cell leukemia]

Between January, 1982, and January, 1992, a total of 112 patients with adult T-cell leukemia (ATL) and 109 patients with non-Hodgkin's lymphoma (NHL) were admitted to our hospital. They were studied for their infectious complications. Infectious complications were seen in 90 patients (80.4%) with ATL, and 51 patients (46.8%) with NHL (p < 0.001). Documented infections were seen in 70 patients (62.5%) with ATL, and 30 patients (27.5%) with NHL (p < 0.001). Pneumonia (p < 0.005), skin infections (p < 0.05), Pneumocystis carinii pneumonia (p < 0.05), fungal infections (p < 0.05), cytomegalovirus infections (p < 0.05) and herpes simplex virus infections (p < 0.01) were identified infections at high risk for patients with ATL. Tuberculosis, listeriosis and salmonella infections were seen only in patients with ATL.     

Similar rearrangements of T-cell receptor beta gene in cell lines and uncultured cells from patients with large granular lymphocyte leukemia

We established interleukin-2-(IL-2) dependent cell lines from three patients with large granular lymphocyte (LGL) leukemia. Phenotypic analysis demonstrated retention of the CD3+, CD8+ phenotype that was observed in the original leukemic LGL. Unique rearrangements of T-cell receptor beta gene occurring in uncultured leukemic LGL, were also found in cell lines, which suggests that the cell lines were derived from the original leukemic LGL clone in each case.