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Expression of Bcl‐2 in feline lymphoma cell lines
Author(s) -
Kano Rui,
Sato Eimi,
Okamura Tomotaka,
Watanabe Shinichi,
Hasegawa Atsuhiko
Publication year - 2008
Publication title -
veterinary clinical pathology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.537
H-Index - 51
eISSN - 1939-165X
pISSN - 0275-6382
DOI - 10.1111/j.1939-165x.2008.00013.x
Subject(s) - monoclonal antibody , microbiology and biotechnology , lymphoma , biology , peripheral blood mononuclear cell , canine lymphoma , antibody , cell culture , recombinant dna , monoclonal , apoptosis , virology , gene , immunology , in vitro , biochemistry , genetics
Background: The Bcl‐2 gene is a member of the rapidly expanding Bcl‐2 family of genes that regulate apoptosis. Bcl‐2 has been shown to repress cell death triggered by a diverse array of stimuli, including chemotherapy and gamma irradiation. Objective: The purpose of this study was to determine feline Bcl‐2 expression level in feline lymphoma cells using an immunoblot assay with anti‐human and anti‐canine Bcl‐2 monoclonal antibodies. Methods: About 708 base pairs containing the coding sequence of the feline Bcl‐2 gene were transformed into Escherichia coli . The recombinant Bcl‐2 was used as a positive control for an immunoblot assay using mouse monoclonal antibodies against human and canine Bcl‐2 . An immunoblot assay using the monoclonal antibodies was carried out to determine the level of feline Bcl‐2 expression in lymphoma and lymphocytic leukemia cell lines. Results: The recombinant feline Bcl‐2 protein produced in E . coli had a molecular weight of about 26 kDa and was detected by immunoblot assay by using anti‐human Bcl‐2 mouse monoclonal antibody. Feline Bcl‐2 expression was high in lymphoma cell lines (FL‐74‐UDC‐1 and FT‐1) and low in the cell line from peripheral blood mononuclear cells from a healthy cat (FeTJ‐1) but not low in freshly isolated peripheral blood mononuclear cells from a healthy cat. The anti‐human Bcl‐2 mouse monoclonal antibody was found to cross‐react with feline Bcl‐2. Conclusions: These results confirm the expression of Bcl‐2 in T‐cell lymphoma cell lines and indicate that it is suitable to detect feline Bcl‐2 using an immunoblot assay. Pending further evaluation, Bcl‐2 expression might be useful in the differential diagnosis of feline tumors.