Sialyl Residues in Hepatitis B Antigen: their Role in Determining the Life Span of the Antigen in Serum and in Eliciting an Immunological Response

Abstract

Summary Hepatitis B surface antigen was adsorbed to insolubilized sialic acid-specific haemagglutinin isolated from the haemolymph of Limulus polyphemus. Treatment of the antigen with Vibrio cholerae neuraminidase (EC 3.2.1.18) resulted in the release of sialic acid and in an increase of the isoelectric point from pH 4.35 (for subtype ad) or 4.9 (for subtype ay) to pH 5.45. Treated, but not untreated, antigen incorporated [14C]-sialic acid when incubated at 37 °C with sialyl transferase (EC 2.4.99.1) and cytidine-5′-monophosphate-[14C]-sialic acid. The major portion of [14C]-sialic acid was linked to a glycoprotein with an apparent mol. wt. of 26 × 103. De-sialylated antigen had a drastically reduced in vivo life span in rabbit plasma and elicited a higher humoral antibody response than intact antigen (subtype ad). Antigen-stimulated proliferation of lymphocytes, measured 3 months after immunization, was observed only with cells from rabbits injected with neuraminidase-treated antigen.