Production of antisera against the enterotoxin of Bacteroides fragilis and their use in a cytotoxicity neutralization assay of HT-29 cells.
To study the enterotoxin of Bacteroides fragilis, the colon carcinoma cell line HT-29 was used in a standard cytotoxicity assay. We produced high-titer neutralizing antisera in rabbits and goats against both crude and purified toxin and developed a cytotoxicity neutralization assay for use in confirming enterotoxin activity in culture filtrates and stools. The neutralization titers of the antisera on the colon carcinoma cell line HT-29 ranged from 1,600 to 2,400. In an antibody screening enzyme-linked immunosorbent assay, titers ranged from 10(4) to 10(5). The antisera produced against the highly purified toxin also neutralized the enterotoxic activity of the toxin and were monospecific by immunoelectrophoresis.
Van Tassel
R L
RL
Department of Biochemistry and Anaerobic Microbiology, Virginia Polytechnic Institute and State University, Blacksburg 24061-0305, USA.
Lyerly
D M
DM
Wilkins
T D
TD
D000818:Animals; D001441:Bacteroides fragilis / Q000276:immunology* / Q000302:isolation & purification; D003601:Cytotoxicity Tests, Immunologic; D004768:Enterotoxins / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D006041:Goats; D019073:HT29 Cells / Q000276:immunology; D006801:Humans; D007122:Immunoelectrophoresis; D009500:Neutralization Tests; D011817:Rabbits
D004768:Enterotoxins