Biological activity of mutants of human tumour necrosis factor-alpha.
Point mutations in different regions of the tumour necrosis factor-alpha (TNF-alpha) molecule influence anti-tumour cytotoxic/cytostatic activities as well as haemorrhagic tumour necrosis, tumour regression and lethal toxicity in mice. Mutations in the C-terminal region in positions 150 and 155 markedly decrease cytotoxicity for murine L929 fibroblasts and human MCF7 mammary carcinoma cells. Competitive binding experiments with 125I-labelled TNF-alpha revealed that the loss of cytotoxicity is caused by a loss of target cell binding. In contrast to the reduced activity against L929 and MCF7 cells, neither binding to nor cytostatic activity against the human myeloid leukaemia cell lines HL60 and U937 are affected. This target cell type-dependent behaviour is probably due to the fact that L929 and MCF7 cells express different types of TNF receptor compared with myeloid leukaemia cells. While a mutation in position 127 decreases the overall activity of TNF-alpha, a deletion of four N-terminal amino acids does not reduce biological activity. In vivo the TNF mutants differed in their anti-tumour effects and lethal toxicity, but a segregation of anti-tumour activity and toxicity was not observed.
Kircheis
R
R
Max Delbrück Center for Molecular Medicine, Berlin-Buch, Germany.
Milleck
J
J
Korobko
V G
VG
Shingarova
L N
LN
Behnke
D
D
Schmidt
H E
HE
D000595:Amino Acid Sequence; D000818:Animals; D001667:Binding, Competitive / Q000187:drug effects; D001943:Breast Neoplasms / Q000473:pathology / Q000628:therapy; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D003602:Cytotoxicity, Immunologic / Q000187:drug effects; D005347:Fibroblasts / Q000187:drug effects; D006801:Humans; D007928:Lethal Dose 50; D007948:Leukemia, Monocytic, Acute / Q000473:pathology / Q000628:therapy; D015473:Leukemia, Promyelocytic, Acute / Q000473:pathology / Q000628:therapy; D051379:Mice; D008969:Molecular Sequence Data; D009154:Mutation; D009369:Neoplasms / Q000188:drug therapy* / Q000473:pathology; D012689:Sequence Homology, Nucleic Acid; D014407:Tumor Cells, Cultured; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics / Q000494:pharmacology* / Q000633:toxicity
D014409:Tumor Necrosis Factor-alpha