Properties of Nine Continuous B‐Cell Lines Established from Enzootic Bovine Leukosis Tumors

Abstract

We established 9 cell lines from 63 tumor cases of enzootic bovine leukosis and studied their properties. Cells of all lines formed small clumps and floated in culture medium, indicating growth. Four of the 9 cell lines were surface immunoglobulin (S1g)-positive, but the remaining 5 line cells were negative for Slg or, if SIg was detected, the percentage of SIg-positive cells was very low. Tests for the properties of the cells with monoclonal antibodies to lymphocytes revealed that the established line cells are B-lymphocytes. Morphological observation also revealed that they had the morphology of B-lymphoblastic cell. The results of E and EAC rosette assay were negative, but 6 of 8 cell lines were positive for EA rosetting. All the 9 cell lines reacted with MoAb C-143, which recognizes the tumorassociated antigen (TAA) of the EBL tumor cell. All 9 cell lines produced bovine leukosis virus (BLV). These results suggest that the 9 cell lines are tumor cells derived from B-lymphocytes of EBL. a) Number of BLV positive dishes; number of dishes tested.

Antitumor Effect of Diphtheria Toxin A‐Chain Gene‐containing Cationic Liposomes Conjugated with Monoclonal Antibody Directed to Tumor‐associated Antigen of Bovine Leukemia Cells

“…It has been reported that about 90% of KU-17 cells showed a positive reaction with c143 monoclonal antibody. 33) In this study, in addition, we found that cationic liposomes coupled with monoclonal antibody were more effective in gene transfer to lymphoid cells than the uncoupled cationic liposomes (Fig. 1).…”

“…KU-17 cells have B cell surface markers and TAA identified by c143 on the cell surface, and produce BLV. 33) P3U1 and C8 cells do not possess TAA recognized by c143 on the cell surface. KU-17 and P3U1 cells were maintained in RPMI-1640 medium (Nissui Pharmaceutical Co., Ltd., Tokyo) supplemented with 10% fetal calf serum and antibiotics.…”

“…4,35) The bovine lymphoid cell line BLSC-KU-17, derived from an EBL tumor, has TAA on its cell surface and about 90% of the cells react with c143 monoclonal antibody in vitro. 33) To confirm the specific targeting of c143 monoclonal antibody to TAApositive cells in vivo, 125 I-labeled c143 monoclonal antibody F(ab′) 2 fragment was injected into athymic nude mice bearing BLV-induced bovine lymphoid cells.…”

Characterization of microRNA expression in B cells derived from Japanese black cattle naturally infected with bovine leukemia virus by deep sequencing

“…RNA was extracted from whole blood collected in PAXgene Blood RNA tubes (PreAnalytix, Hombrechtikon, Switzerland) and stored at -70˚C for several months after the blood collection. RNA was also extracted from the bovine B cell leukemia cell line KU-17 [27] with TRIzol Reagent (Invitrogen, Carlsbad, CA, USA). These DNA and RNA extraction procedures were performed according to manufacturer's instructions.…”

Comprehensive Comparison of Novel Bovine Leukemia Virus (BLV) Integration Sites between B-Cell Lymphoma Lines BLSC-KU1 and BLSC-KU17 Using the Viral DNA Capture High-Throughput Sequencing Method

“…The bovine B-cell lymphoma lines BLSC-KU1 and BLSC-KU17 were established from leukemic cells of BLV-infected cattle with EBL. Both cell lines grow as single, free-floating cells that do not attach to a glass surface [ 12 , 13 , 14 ]. These cell lines have B-cell surface marker and tumor-associated antigen on their cell surface [ 12 , 13 , 14 , 15 , 16 ].…”