Production of diphtheria antitoxin antibody in Epstein-Barr virus-induced lymphoblastoid cell lines
Tsuchiya, S.; Yokoyama, S.; Yoshie, O.; Ono, Y.
Journal of Immunology 124(4): 1970-1976
1980
ISSN/ISBN: 0022-1767 PMID: 6245131 Document Number: 162809
Human tonsil lymphocytes were cultured for 48 h with diphtheria toxoid and then infected Epstein-Barr virus to establish lymphoblastoid cell lines (LCL). Diphtheria antitoxin activities in the culture supernatants from these LCL were measured by passive hemagglutination with diphtheria toxoid-coated human red blood cells. Various titers of antitoxin activity were detected in 40 of 52 LCL derived from 4 tonsil specimens. The properties of the antitoxin activity were as follows: the activity was inhibited by the toxoid; it neutralized the toxin; it was inhibited by the anti-.gamma. chain serum; and the Sephadex G-200 gel elution pattern was similar to that of mouse anti-DNP-KLH G antibody. The antitoxin activity produced by the LCL was classified as IgG. The limiting dilution cultures showed that there were at least 10-260 antitoxin-producing cells in 106 lymphoblastoid cells. Serial passaging of the LCL gradually decreased the production of antitoxin antibody, which became undetectable after 2 mo. in most cultures. One cell line (TC-9) continued producing a constant titer of antitoxin antibody until the 110th day after initiation of the in vitro culture. Diphtheria antitoxin, anti-sheep red blood cells, anti-streptolysin O, anti-DNP hapten, and anti-phosphorylcholine antibody activities could be detected in the culture supernatants.