Glycosylation-dependent peptide antigenic determinants of env gp46 HTLV-1.

Human T-lymphotropic virus type 1 (HTLV-I) is a retrovirus associated with adult T-cell leukemia/lymphoma, and the virus infection constitutes a growing public health problem. In a continuing effort to engineer conformationally dependent HTLV-I epitopes that elicit a protective immune response, we have examined the role and functional importance of carbohydrate moieties in specific immune recognition and antibody responses. There have been several reports of the importance of N-linked virus glycosylation in the formation of neutralizing antibodies. Residues 230-257 is predicted to encode two β-turn/loop regions at 240-244 (LYGPN), 248-257 (VPSSSSTPL) and a glycosylation site at N-244 (NVS). We have successfully engineered and synthesized the 233-253 sequence of gp46 of HTLV-1 with and without GlcNAC at Asn244. Circular dichroism spectroscopy and proton NMR showed the presence of β-turn conformation in both peptide constructs. Chimeras of the glycosylated and non-glycosylated epitope with promiscuous T-cell epitope were synthesized and shown to elicit high titered antibodies in rabbits specific for the immunogen (SC1MVF and SC2MVF) and the B cell epitope 233-253. Additionally, antibodies to the glycosylated form of the peptide recognized the HTLV-I envelope precusor in radioimmunoassay precipitation assay and react with HTLV-I whole virus preparations in ELISA.