Open AccessCharacterization of a Novel Human Tumor Necrosis Factor‐α Mutant with Increased Cytotoxic Activity
Author(s) -
Masegi Tsukio,
Kato Arata,
Kitai Kazuo,
Fukuoka Masami,
Ogawa Hiroko,
Ichikawa Yataro,
Nakamura Satoshi,
Watanabe Naoki,
Niitsu Yoshiro
Publication year - 1995
Publication title -
japanese journal of cancer research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.035
H-Index - 141
eISSN - 1349-7006
pISSN - 0910-5050
DOI - 10.1111/j.1349-7006.1995.tb02990.x
Subject(s) - cytotoxic t cell , mutant , tumor necrosis factor alpha , microbiology and biotechnology , recombinant dna , biology , biological activity , cytotoxicity , cell culture , chemistry , in vitro , biochemistry , immunology , gene , genetics
Various novel recombinant human tumor necrosis factor‐α (TNF) mutants were prepared using protein engineering techniques, and their cytotoxic activity was compared with that of the intact form of TNF (intact TNF). Mutant 471 (a TNF mutant molecule with the deletion of 7 amino acids at the amino‐terminal and the substitution of Pro 8 Ser 9 Asp 10 by ArgLysArg) had a 6‐fold higher cytotoxic activity against murine L929 cells. The mutant TNF had an increased ability to bind to TNF receptor on murine L929 fibroblasts cells. A cross‐linking study revealed that mutant 471 had an increased ability to form an active trimer. Mutant 471 also showed higher cytotoxic activity against human KYM myosarcoma cells and human MIA PaCa‐2 pancreatic carcinoma cells. The possible cachectin activity of the mutant was almost the same as that of intact TNF. These results suggest that mutant 471 might be a more promising candidate as an anticancer agent than intact TNF.