Effect of heptakis (2,6-0-dimethyl)beta-cyclodextrin on cell growth and the production of pertussis toxin and filamentous hemagglutinin in Bordetella pertussis

Abstract

Heptakis (2,6-0 dimethyl)beta-cyclodextrin (MeCD) which permits the growth of single colonies of Bordetella pertussis Tohama phase I on Stainer-Scholte medium solidified with agar also enhanced the production of pertussis toxin (PT). More than one hundred times the amount of PT was produced in Stainer-Scholte medium with MeCD in shake culture than was produced in MeCD-free medium. A maximum of 50 mg PT protein was produced per liter of culture broth as estimated by in vitro and in vivo assays. The production of filamentous hemagglutinin (FHA) was several hundred times greater when B. pertussis was grown in shake cultures with MeCD than when growth was in MeCD-free shake cultures. The FHA content of the production medium was confirmed by enzyme-linked immunosorbent assays and electron microscopy. Evaluation of an acellular vaccine containing PT and FHA detoxified with formaldehyde showed that it was protective in the intracerebral challenge mouse potency assay.