Analysis of transformation with epstein‐barr virus and phenotypic characteristics of lymphoblastoid cell lines established from patients with hairy cell leukemia

Abstract

In order to assess the role of Epstein-Barr virus (EBV) in patients with hairy cell leukemia (HCL), we have sought to characterize 1) the ability of EBV to infect and transform hairy leukemic cells in vitro and 2) the phenotypes of cell lines putatively derived from those leukemic cells. Analysis of EBV-induced transformation and the kinetics of Epstein-Barr nuclear antigen (EBNA) induction in leukemic preparations indicated that most leukemic cells were not susceptible to EBV infection but that at least a small subpopulation of leukemic cells could be infected with EBV. Lymphoblastoid cells lines were established after exposure of peripheral blood or splenic cells from HCL patients to B95-8 or QIMR-WIL EBV. Splenic leukemic cell preparations were more sensitive targets for EBV transformation than were peripheral blood cell samples. The newly established cell lines, but not long-established B lines such as Raji, demonstrated high levels of synthesis of p35, (a protein complex expressed abundantly by cells of a subset of HCL patients) and high levels of tartrate-resistant acid phosphatase (an enzyme relatively diagnostic for HCL). Lymphoblastoid lines from one patient with HCL expressed lambda light chains and no kappa chains as did the patient's leukemic cells. Virus expression in these lines showed that HCL-derived lines had spontaneous early antigen (EA) and viral capsid antigen (VCA) expression. Transforming EBV could be rescued from HCL-derived cell lines but not from cord blood-derived lines.

Predilection of a nasopharyngeal carcinoma‐derived isolate of epstein‐barr virus for infection of specific subsets of B lymphocytes

“…Many studies have explored the importance of the stage of B-lymphocyte maturity on EBV transformation by IF using antibodies to cytoplasmic and surface Ig [Sairenji et al, 1983;Katamine et al, 1984;Chan et al, 1986;Gregory et al, 1987;Kubagawa et al, 19891. Chan et al [19861 showed heterogeneity of B-lymphocyte cultures transformed by EBV.…”

“…We have explored the possibility that there might be different target B lymphocytes for the NPC-EBV isolate as compared with two other EBV isolates, B95-8 and AG876, since this isolate was originally obtained from EBV genome-positive NPC epithelial tumor cells and maintained continuously in latently infected epithelial-epithelial hybrid cells in vitro [Takimoto et al, 1985;Sat0 et al, 19861. Many studies have explored the importance of the stage of B-lymphocyte maturity on EBV transformation by IF using antibodies to cytoplasmic and surface Ig [Sairenji et al, 1983;Katamine et al, 1984;Chan et al, 1986;Gregory et al, 1987;Kubagawa et al, 19891. Chan et al [19861 showed heterogeneity of B-lymphocyte cultures transformed by EBV.…”

Differential effect of TPA and n‐butyrate on induction of Ii and EBV antigens in the P3HR‐1 lymphoblastoid cell line

“…In the case of some leukemic cells, Ii might influence Ia function and, thus potentially, the immune response to the leukemia. Hyperexpression of Ii has been defined on cells of a subset of patients with hairy cell leukemia (Spiro et al, 1980(Spiro et al, , 1984a, on prolymphocytic leukemia cells (Spiro et al, 1983),on freshly EBV-transformed lymphoblastoid cell lines , and on n-butyrate-treated Burkitt's lymphoma cell lines which then express EBV antigens (Spiro et al, 1984b). Since p35, coelectrophoresing with Ii in SDS-PAGE, was also induced in P3HR-l EBV-superinfected Raji cells (Casareale et al, 1984), and Ii has been demonstrated in freshly EBV-transformed B cell lines, one could consider Ii to be an EBVinduced antigen.…”

“…Cell viability was determined by trypan blue dye exclusion. Previously described techniques were followed for [ 35S]methionine-labelling of cells, isolation of membranes, preparation of immunoprecipitates and SDS-PAGE (Malenka et al, 1979;Spiro et al, 1983). (Spiro et al, 1984b).…”

Establishment and Characterization of an Epstein-Barr Virus Nuclear Antigen-Negative Hairy Cell Line

“…Since the Ficoll-purified spleen cell popula tion did not consist of pure hairy cells, there is at present no way to decide whether EBV caused transformation of normal lympho cytes or hairy cells. Recent studies showed that only a subpopulation of hairy' cells from a spleen preparation was susceptible to delib erate EBV infection [Sairenji et al, 1983]. Subpopulations of hairy cells have previously been suggested on the basis of different crite ria [Golomb et al, 1978;Spiro et al, 1980].…”