OvB20, an Onchocerca volvulus-cloned antigen selected by differential immunoscreening with vaccination serum in a cattle model of onchocerciasis

Abstract

A cDNA of Onchocerca volvulus has been isolated by differential immunoscreening of an adult worm expression library using sera raised in cattle against the related species, O. lienalis. It was selected because of its recognition by antibodies from cattle immunized with irradiated third-stage (L3) larvae and not by antibodies from animals infected with non-irradiated larvae. The original 311-bp clone was used to isolate a 1478-bp cDNA. Designated OvB20, this codes for 460 amino acid residues, hybridizes with a ∼ 1.6 kBp transcript and appears to be transcribed from a filarial-specific, single copy gene. It is expressed in developing stages from embryo to L4 larva, but not in the adult. The product of OvB20 appears to undergo co- or post-translational processing: in vitro transcription and translation give rise to a polypeptide consistent with the deduced amino acid sequence (approximately 52 kDa), whilst products of 52 and 65 kDa are detected in larvae by immunoblotting and following in vitro translations to which exogenous microsomes have been added. A 42-kDa protein was also detected in all in vitro translations. No homologous genes were found in the computer databases, although there are regions of weak sequence similarity with C-reactive proteins. The functional role of OvB20 may warrant further attention, as it has recently been shown that the recombinant protein confers host protection against a related rodent filaria following active immunization (Taylor, M.J., Abdel-Wahab, N., Wu, Y., Jenkins, R.E. and Bianco, A.E. (1995) Onchocerca volvulus larval antigen, OvB20 induces partial protection in a rodent model of onchocerciasis. Infect. Immun. 63, 4417–4422).