Progression of a human B cell chronic lymphocytic leukemia line in nude mice

Abstract

Subcutaneous (s.c.) inoculation of the 85-4LN subline, derived from a lymph nodal metastasis of the Epstein-Barr virus (EBV) transformed human chronic lymphocytic leukemia (CLL) B cell line, EBV-CLL (1), produced progressively growing lethal tumors in 31/35 nonirradiated (88.6%) and 22/25 (88%) of whole-body irradiated (440 rad) nude mice. In contrast, EBV-CLL(1) could produce progressive tumors only in irradiated nude mice. All 85-4LN cells had Epstein-Barr virus nuclear antigen and reacted with pan B and anti-la antibodies. The morphology and ultrastructural features was consistent with the lymphoblastoid nature of the cells. In all s.c. tumor bearing mice, there was enlargement of the spleen and draining lymph nodes. Karyological studies revealed human cells in the spleen and draining nodes in all the mice investigated. Metastases in nonlymphoid organs were seen in 1/8 irradiated and 8/12 nonirradiated mice. The subline contained 77% cells with 47,XY, +12 and 23% cells with 45,XY karyotype. The clone with trisomy 12 did not have any growth advantage either in s.c. transplants or in splenic/lymph nodal metastases. Treatment with the maximum permissible doses of methotrexate (MTX) or chlorambucil (CBL) revealed xenografts to be more sensitive to MTX than CBL. A clone with a 1g+ marker, i.e., 46,XY,Dup(1) (q11----q32) appeared to be associated with resistance to CBL. We have not seen any previous report on the growth and dissemination of human CLL B cells in nonirradiated nude mice. The 85-4LN subline, thus, provides a model for studying the progression, dissemination and therapeutic response of human CLL-B cells.

Inhibition of human B‐cell chronic lymphocytic leukemia by a monoclonal antibody in xenograft models

“…The origin of the EBV-transformed human B-cell CLL line EBV-CLL-1 from a CLL Rai stage I1 patient (Lee et al, 1986) and of its subclone Dlo.' with partial duplication of chromosome lq, i.e., 46, XY, dup(1) (qll + q32), has been described (Ghose et al, 1988(Ghose et al, , 1990. HL6", a human promyelocytic leukemia cell line, Yac, a mouse lymphoma cell line, and ATCC TIB9, a non-specific mouse IgG1-producing myeloma cell line, were purchased from the ATCC (Rockville, MD).…”

“…Chromosomal analysis of tumor cells harvested from ascites and solid tumors, as well as of the parent Dlo.l cells, was carried out as described (Ghose et al, 1988). Briefly, for harvesting metaphases, aliquots of dissociated tumor cells obtained from mouse ascites or solid tumors were first exposed to Colcemind (0.05 Fgiml, Sigma) for approximately 2 hr and then to a 0.075% M KCI solution for 20 min.…”

“…The chemosensitivity of tumor cells to MTX (Sigma) and ADR (Adria, Columbus, OH) was determined as described (Ghose et al, 1988). Briefly, aliquots of 3 x lo4 tumor cells in 100 ~1 of medium were plated in each well of a 96-well culture plate and incubated overnight.…”

“…We have reported progressive lethal growth along with lymph-nodal and splenic metastasis of S.C. xenografts of an EBV-transformed human B-cell CLL line in both irradiated (Lee et al, 1986) and non-irradiated nude mice, and have isolated a clone of this CLL line, clone Dlo.1, that after S.C. inoculation into nude mice produces progressively growing S.C. tumors with multiple organ metastasis (Ghose et al, 1988(Ghose et al, , 1990. However, there still remains a need for experimental models of CLL that have a close resemblance to the clinical disease or that allow a comparatively rapid evaluation of therapeutic agents.…”

Monoclonal Antibodies Against Epstein-Barr Virus Transformed B Lymphocytes from a CLL Patient

“…Animals maintained under germfree conditions were examined, and tumors were measured three times a week (13).…”