ESTABLISHMENT AND CHARACTERIZATION OF AN EBNA-NEGATIVE HUMAN LYMPHOMACELL-LINE (BHL-89)

Background. The aim of the work was to establish human malignant lymph omas in culture, in order to study the biological characteristics and drug sensitivity of lymphomas of human lymphoid origin. Materials and Methods. Lymph nodes of patients were explanted and kept in cultures u sing conventional tissue culture methods. Cytogenetic methods were use d for karyotype analysis. Clonogenic assay was applied to test drug se nsitivity. The tumorigenic capacity of the cells was determined by ino culating them into immunosuppressed mice. Immunological and other mark ers were examined with conventional techniques. Results. A cell line, BHL-89, was established in culture from a patient with B-cell type non -Hodgkin's malignant lymphoma. Cells started to grow after a few days without a feeder layer in stationary suspension. The population doubli ng time was 48 h. The cells were hyperploid, and nonrandom aberrations were +1, -15, +14q+. Cloning efficiency in soft agar was found to be as high as 50-60%. The cells expressed markers characteristic of early B cells. The BHL-89 cells were Epstein-Barr nuclear antigen (EBNA) ne gative. They produced tumors when 10(7) cells were injected into immun osuppressed mice. The cells were sensitive to dibromodulcitol (Elobrom ol), an alkylating antitumor drug, and resistant to the phorbol ester TPA; Conclusions. The established EBNA-negative BHL-89 cell line has a few unique characteristics, e.g. rapid establishment without feeder c ells, origin from the lymph node of an adult patient, high clonogenici ty in soft agar, and resistance to TPA. The cell line is suitable for studying the nature of B lymphomas and testing compounds against lymph oproliferative disorders.