SIMULTANEOUS PRODUCTION OF NATURAL HUMAN-TUMOR NECROSIS FACTOR-ALPHA, FACTOR-BETA AND INTERFERON-ALPHA FROM BALL-1 CELLS STIMULATED BY HVJ

  • 1 June 1988
    • journal article
    • research article
    • Vol. 7  (2) , 175-185
Abstract
B-cell human lymphoblastoid cell line, BALL-1, produced interferon-.alpha. (IFN-.alpha.) and cytotoxic factors by the stimulation of hemagglutinating virus of Japan (HVJ). These cytotoxic factors were identified to be tumor necrosis factor-.alpha. (TNF-.alpha.) and tumor necrosis factor-.beta. (TNF-.beta.) on the basis of molecular level analysis. In the kinetic study, it was found that the maximum production of IFN-a was reached at 16 .apprx. 20 hr and that of TNF-.alpha. was at 10 .apprx. 16 hr after the stimulation by HVj, whereas the maximum production of TNf-.beta. was at 40 .apprx. 50 hr, suggesting that the regulation of TNF-.alpha. and TNF-.beta. genes are different. These cytokines were purified by monoclonal antibody linked column chromatography, respectively. IFN-.alpha. was further separated into 9 subspecies on the basis of the difference of isoelectric point by FPLC chromatofocusing method. From the N-terminal amino acid sequence analyses of the 7 species of 9 subspecies, these subspecies were classified into 3 subtypes according to the Weissmann''s notation. On the other hand,the amino acid sequences of the tryptic peptides of purified TNf-.alpha. and TNF-.beta. coincided well with the parts of the complete amino acid sequences which has been deduced from complementary DNA of TNF-.alpha. and TNF-.beta. of BALL-1 cell.