Insertion of a foreign sequence on capsid surface loops of human papillomavirus type 16 virus-like particles reduces their capacity to induce neutralizing antibodies and delineates a conformational neutralizing epitope.
The aims of this study were to generate chimeric human papillomavirus (HPV)-16 L1 virus-like particles (VLPs) in order to identify immunogenic domains and conformational neutralizing epitopes, and to characterize the regions where a foreign epitope could be introduced. We hypothesized that these regions could be on L1 protein loops since they are exposed on the surface of VLPs. The aims of this study were achieved by mutating HPV-16 L1 proteins. Six amino acids encoding for the epitope 78-83 (DPASRE) of the hepatitis B core (HBc) antigen were introduced within the different loops of the L1 protein at positions 56/57, 140/141, 179/180, 266/267, 283/284 or 352/353. All these chimeric L1 proteins were capable of self-assembly into VLPs. The antigenicity and immunogenicity of some of these VLPs were reduced compared to the levels observed with wild-type VLPs. All were nevertheless able to induce neutralizing antibodies. VLPs with insertion at position 266/267 induced lower levels of neutralizing antibodies, suggesting the involvement of residues situated on FG loop in L1 neutralizing epitopes. All the chimeric L1 proteins except the one with insertion at position 56/57 were also able to induce anti-HBc antibodies, thus suggesting exposure of the HBc epitope on the VLP surface. Taken together, our findings indicate the possibility of designing HPV-derived vectors that are less immunogenic and suggest positions for insertion of defined immune epitopes or cell ligands into L1 protein to be exposed on the surface of VLPs.
Laboratoire de Virologie Moléculaire, INSERM EMIU 00-10 and USC INRA, IFR 82 Transposons et Virus, Faculté des Sciences Pharmaceutiques, 31 Avenue Monge, 37200 Tours, France.
Jean-Rémy
JR
Sadeyen
Sylvie
S
Tourne
Marina
M
Shkreli
Pierre-Yves
PY
Sizaret
Pierre
P
Coursaget
D001483:Base Sequence; D036022:Capsid Proteins / Q000737:chemistry* / Q000235:genetics; D000939:Epitopes / Q000737:chemistry; D006801:Humans; D009500:Neutralization Tests; D009838:Oligodeoxyribonucleotides; D027383:Papillomaviridae / Q000235:genetics* / Q000276:immunology
D036022:Capsid Proteins; D000939:Epitopes; D009838:Oligodeoxyribonucleotides