title: Molecular cytogenetic analysis of feline leukemia virus insertions in cat lymphoid tumor cells.
abstract: This study was conducted to map the acquired proviral insertions in the chromosomal genome of feline lymphoid tumors induced by feline leukemia virus (FeLV). Chromosome specimens of the lymphoid tumor-derived cell lines and normal cat lymphocytes were subjected to fluorescence in situ hybridization and tyramide signal amplification, using an exogenous FeLV-A genome as a probe. Specific hybridization signals were detected only on the metaphase chromosomes of the tumor cells. Poisson's distribution-based statistics indicated that 6 chromosomal loci in each cell line showed FeLV integration. In the examination of metaphase chromosomes of FL-74, FT-1 and KO-1 cells, significant signals were detected on B2p15-p14, B2q11, D1p14, E1p14-p13, E1q12 and F2q16; A2p23-p22, B2p15-p14, B4p15-p14, D4q23-q24, E1p14-p13 and E2p13-p12; and A2p22, A3q22, B1p13, B1q13, D1p13 and D3p15-p14, respectively. Consistently, Southern blot hybridization using an FeLV LTR-U3 probe specific for exogenous FeLV revealed the presence of at least 6 copies of exogenous FeLV proviruses at different integration sites in each cell line. These results indicate that there may be common FeLV integration sites at least in A2p22 and B2p15-p14. The cytogenetic analysis used in this study can promptly screen FeLV insertions and provide tags for identifying the novel common integration site.
authors.affiliation: Department of Veterinary Internal Medicine, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan. afujino@mail.ecc.u-tokyo.ac.jp
authors.forename: Yasuhito
authors.initials: Y
authors.lastname: Fujino
authors.forename: Hitoshi
authors.initials: H
authors.lastname: Satoh
authors.forename: Koichi
authors.initials: K
authors.lastname: Ohno
authors.forename: Hajime
authors.initials: H
authors.lastname: Tsujimoto
mesh_terms: D000818:Animals; D015139:Blotting, Southern; D002415:Cats; D045744:Cell Line, Tumor; D002875:Chromosomes / Q000821:virology; D020732:Cytogenetic Analysis; D017404:In Situ Hybridization, Fluorescence / Q000379:methods; D007621:Karyotyping; D005256:Leukemia Virus, Feline / Q000235:genetics*; D008223:Lymphoma / Q000821:virology*; D016254:Mutagenesis, Insertional; D011533:Proviruses / Q000235:genetics*; D016662:Virus Integration