Establishment and characterization of five human malignant mesothelioma cell lines derived from pleural effusions

Abstract

Malignant mesothelioma (MM) is an aggressive tumour of the serosal cavities which is associated with exposure to asbestos. Studies of this tumour have been limited by a paucity of well-characterized human MM cell lines. In this study, 5 human MM cell lines were established from pleural effusions of patients with this malignancy. All 5 patients were males with known crocidolite asbestos exposure, who had received no treatment for their disease and in whom the diagnosis was confirmed by cytology, histology and electron microscopy (EM). These lines have been in culture from 11 to 25 months, and all of them for more than 18 passages. The appearance of the cells in culture was extremely varied; in 3 of the lines they were spindle-shaped with few vacuoles (JU77, LO68 and ONE58); in 1 line they had a thick, stellate shape with vacuoles (NO36) and in 1 they were very pleomorphic in both shape and size with irregular membranes and numerous vacuoles [DeH128 (M)]. Upon reaching confluence, cells in 3 of the 5 lines assumed the cobblestone-like pattern characteristic of epithelial-type cells, whereas in the other 2 (LO68 and ONE58) they remained spindle-shaped. All 5 lines demonstrated a loss of contact inhibition (i.e., piling) at confluence. Minimum doubling times varied significantly from 18 hr (JU77) to more than 30 hr [DeH128 (M)]. Cytological examination showed characteristic mesothelial/mesothelioma morphology, and epithelial membrane antigen (EMA) and cytokeratin were demonstrated in cells from all 5 lines. These cells lacked CEA and epithelial mucin. The presence of cell junctions, glycogen and numerous long, thin, branching microvilli was readily demonstrable by EM. All lines had abnormal karyotypes, with the modal chromosome number varying from 40 to 80. Variable chromosome numbers, numerous structural rearrangements and unrecognizable marker chromosomes were readily observed; however, the only consistent change seen was del 6q21 in 4 of the 5 lines. The establishment of these 5 cultured human MM cell lines now provides an opportunity for comparative study of several aspects of the biology of MM in vitro as well as screening new treatment modalities.

Dual Control of Antitumor CD8 T Cells through the Programmed Death-1/Programmed Death-Ligand 1 Pathway and Immunosuppressive CD4 T Cells: Regulation and Counterregulation

“…We used a mouse model of mesothelioma (15,17) to assess the role of PD-1-mediated T cell inhibition in natural tumor-induced immune responses (i.e., without additional therapy). First, we evaluated the expression patterns of PD-L1 on AB1-HA mesothelioma cells.…”

“…However, it has recently been shown that CD25 ϩ regulatory T cells are also under the control of PD-1, i.e., PD-L1 blockade enhanced the suppressive function of regulatory T cells (14). Therefore, the objective of the present study was to investigate the role of PD-1 in tumor-induced immunosuppression in a mouse model of malignant mesothelioma (15), in which immunosuppression depends in part on regulatory T cells (16). To study this, we have used the AB1-hemagglutinin (HA) mesothelioma tumor cell line that was obtained by transfecting asbestos-induced AB1 mesothelioma cells (15) with the infuenza virus HA gene as model tumor Ag (17).…”

“…Therefore, the objective of the present study was to investigate the role of PD-1 in tumor-induced immunosuppression in a mouse model of malignant mesothelioma (15), in which immunosuppression depends in part on regulatory T cells (16). To study this, we have used the AB1-hemagglutinin (HA) mesothelioma tumor cell line that was obtained by transfecting asbestos-induced AB1 mesothelioma cells (15) with the infuenza virus HA gene as model tumor Ag (17). Expression of the HA gene does not impact on the immunogenicity of the tumor (18).…”

A Proteomic Analysis of the Malignant Mesothelioma Secretome Using iTRAQ

“…Six previously established MM cell lines, JU77, LO68, ONE58, OLD1612, JO38 and GAY2911, were maintained as previously described (5). The cell lines and the patient demographics from which the cell lines were derived are described in Table I.…”

p53 autoantibodies in patients with malignant mesothelioma: stability through disease progression

“…The human MM cell lines, JU77, LO68, NO36, ONE58 and STY51 were all derived from the pleural fluid of patients with disease (Manning et al, 1991). The human lung adenocarcinoma, A549, and the human colon cancer line HT29 were purchased from the American Type Culture Collection (Manassas, VA).…”