Characterization of a cytotoxic factor produced by rat bladder cancer cell line: its identity to TNF alpha.
Clone C19 derived from a rat bladder cancer cell line BC31ad constitutively released cytotoxic factor RCF. The serum-free C19-conditioned medium was partially purified by DEAE-Sephacel, Sephacryl S-200 and MonoQ-FPLC chromatographies to the specific activity of 6.4 x 10(6) U/mg protein (LM assay). RCF was a 17-kDa protein with pI 4.6-5.0. Anti-RCF serum neutralized the cytotoxic activity of rat and mouse, but not human, tumor necrosis factor-alpha (TNF alpha) as well as RCF. The partial amino acid sequence of RCF was homologous with the sequence 43-58 of rat TNF alpha. RCF and rat TNF alpha showed a similar cytotoxic spectrum against several tumor cells. Furthermore, they induced a marked proliferation of mouse T cells in combination with a suboptimal dose of ConA or anti-CD3 antibody, however, human TNF alpha almost not. Taken together, RCF was substantially regarded as rat TNF alpha.
Department of Hygienic Chemistry, Tohoku University, Sendai, Japan.
H
H
Hojo
A
A
Kaneko
K
K
Nanya
N
N
Kayagaki
Y
Y
Hashimoto
D000595:Amino Acid Sequence; D000818:Animals; D003602:Cytotoxicity, Immunologic; D055646:Killer Factors, Yeast; D008213:Lymphocyte Activation / Q000187:drug effects; D051379:Mice; D008809:Mice, Inbred C3H; D008969:Molecular Sequence Data; D008970:Molecular Weight; D014176:Protein Biosynthesis; D011506:Proteins / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification; D051381:Rats; D011912:Rats, Inbred ACI; D013601:T-Lymphocytes / Q000276:immunology; D014407:Tumor Cells, Cultured; D014409:Tumor Necrosis Factor-alpha / Q000737:chemistry* / Q000276:immunology; D001749:Urinary Bladder Neoplasms / Q000378:metabolism*
D055646:Killer Factors, Yeast; D011506:Proteins; D014409:Tumor Necrosis Factor-alpha