Immune system status was characterized in patients with hairy cell leukemia (HCL) with respect to explaining their chronic or recurrent infections with Epstein-Barr virus. Measures of cellular immune responsiveness for a group of 11 HCL patients were, in general, decreased when expressed as the proportion of tested patients with values less than 2 S.D. below mean values for a group of 17 healthy adults: T-cell enumeration, seven of 13; mitogen responsiveness of phytohemagglutinin, 10 of 11; concanavalin A, 10 of 11; pokeweed mitogen, 10 of 11; B-cell responsiveness by anti-immunoglobulin immunobead stimulation, two of six; responsiveness to streptolysin O antigen, four of seven; mixed-lymphocyte reaction, six of seven; natural killer cell activity, six of eight. Specific immunity to Epstein-Barr virus was measured by complement-independent, antibody-mediated virus neutralization (mean index for HCL patients being 56% of control value) and complement-dependent virus neutralization (98% of control value). We concluded that, in spite of depressed levels of immune responses measured with general, cellular assays, functional levels of complement-dependent virus-neutralizing antibody were present in these HCL patients.

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.

Morphological and biochemical differences were demonstrated between prolymphocytic leukemia cells obtained from the spleen and peripheral blood of one patient. Peripheral blood prolymphocytes had consistently smaller nuclear-cytoplasmic ratios than did splenic prolymphocytes. Percoll gradient-purified prolymphocytes from the spleen synthesized abundant amounts of some membrane proteins which were hardly expressed by peripheral blood prolymphocytes. Peripheral blood prolymphocytes did not change their expression of membrane proteins during three days in culture. These findings are consistent with the view that prolymphocytic leukemia cells from the spleen exist, on the average, at an earlier stage of differentiation than do circulating leukemic cells, and that peripheral blood leukemic cells are frozen at a specific phase of differentiation.