Molecular and Biological Properties of an Immunopotentiating Complex Polysaccharide Adjuvant Produced by a Gliding Bacterium.
Abstract
Gliding bacteria adjuvant (GBA) is composed of macromolecular substances secreted or shed into the growth media of a newly described Cytophaga species. We had established prior to the onset of this contract that GBA strongly augments the in vitro generation of humoral immunity by mouse spleen cells and that it is mitogenic for murine spleen cells. Since GBA is very water soluble, its absorbance at 280 nm is low, and its biological activities are stable to boiling, we hypothesized that GBA was a polysaccharide or a complex of polysaccharides. Our principal objectives during the first year of this contract were to develop methods for large scale production and purification of GBA, to initiate studies of its physical and chemical properties, to prepare a panel of monoclonal antibodies against GBA for future studies defining its biologically significant epitopes and to extend studies of its biological activities. Large scale production and purification have been essentially achieved using protocols described in this report. Knowledge of the biological activities of GBA for mammalian cells was extended this year. Titration curves with the most highly purified preparations of GBA were performed with assays measuring in vitro adjuvanticity, mitogenicity, B cell polyclonal activation and induction of the murine macrophage cell line, WEHI 265, to secrete IL-1. Studies on the mechanism of action of GBA were initiated.
Document Details
- Document Type
- Technical Report
- Publication Date
- Dec 13, 1985
- Accession Number
- ADA162664
Entities
People
- Robert I. Mishell
- William R. Usinger
Organizations
- University of California, Berkeley