Establishment and characterization of an EBNA-negative human lymphoma cell line (BHL-89)

The aim of the work was to establish human malignant lymphomas in culture, in order to study the biological characteristics and drug sensitivity of lymphomas of human lymphoid origin.

Lymph nodes of patients were explanted and kept in cultures using conventional tissue culture methods. Cytogenetic methods were used for karyotype analysis. Clonogenic assay was applied to test drug sensitivity. The tumorigenic capacity of the cells was determined by inoculating them into immunosuppressed mice. Immunological and other markers were examined with conventional techniques.

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 doubling time was 48 h. The cells were hyperploid, and non-random 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) negative. They produced tumors when 10(7) cells were injected into immunosuppressed mice. The cells were sensitive to dibromodulcitol (Elobromol), an alkylating antitumor drug, and resistant to the phorbol ester TPA.

The established EBNA-negative BHL-89 cell line has a few unique characteristics, e.g. rapid establishment without feeder cells, origin from the lymph node of an adult patient, high clonogenicity in soft agar, and resistance to TPA. The cell line is suitable for studying the nature of B lymphomas and testing compounds against lymphoproliferative disorders.