Immunological studies in ulcerative colitis: Extraction, fractionation, and immunological characterization of germ-free rat colon antigen

Abstract

Intestinal mucins from germ-free rats contained antigens reactive with sera from patients with ulcerative colitis, in addition to human blood group A- and H-like antigens. A crude antigen extract was obtained by phenol-water extraction at 65 °C. Two intestinal glycoproteins were purified from the extract by fractionated ethanol precipitation, ion-exchange chromatography, and gel filtration. The two glycoproteins (2aI and 4aIIb) were homogeneous in regard to electrical charge and molecular size. Both were glycoproteins of the blood group substance type. Component 2aI was very rich in N-acetylgalactosamine and threonine and low in N-acetylglucosamine and sialic acid(s). It had strong blood group A-like activity, weak blood group H activity, and no colon antigen activity as defined by patients' sera. Component 4aIIb was rich in sialic acid(s). About 40% of the sera from patients with ulcerative colitis reacted with this component. No blood group A- or H-like activity could be demonstrated. Colon antigen activity was sensitive to periodate oxidation, but resistant to boiling at neutral pH. It was very sensitive to acid hydrolysis. In fact, colon antigen activity was significantly reduced when subjected to weak acid hydrolysis under conditions which only appeared to release sialic acids.