A disulfide-bound HIV-1 V3 loop sequence on the surface of human rhinovirus 14 induces neutralizing responses against HIV-1.
An immunogenic sequence from the V3 loop of the MN isolate of human immunodeficiency virus type 1 (HIV-1), His-Ile-Gly-Pro-Gly-Arg-Ala-Phe, was transplanted onto a surface loop of the VP2 capsid protein of human rhinovirus 14. To optimize for virus viability and immunogenicity of the transplanted sequence, the HIV sequence was flanked by (1) a cysteine residue that could form a disulfide bond and (2) randomized amino acids (in either of two arrangements) to generate numerous presentations of the Cys-Cys loop. The location for engineering in VP2 was chosen by searching the geometries of disulfide-bound loops in known protein structures. A model for the structure of the transplanted V3 loop sequence was developed using molecular dynamics and energy minimization calculations. Proteolytic digestion with and without reducing agent demonstrated the presence of the disulfide bond in the chimeric virus examined. Monoclonal and polyclonal antibodies directed against the V3 region of the HIV-1MN strain potently neutralized two chimeric viruses. Guinea pig antisera against two chimeric viruses were able to neutralize HIV-1MN and HIV-1ALA-1 in cell culture. The ability of chimeric viruses to elicit antibodies capable of neutralizing the source of the transplanted sequence could be favorable for vaccine development.
Zhang
A
A
Center for Advanced Biotechnology and Medicine and Department of Chemistry, Rutgers University, Piscataway, NJ 08854, USA.
Geisler
S C
SC
Smith
A D
AD
Resnick
D A
DA
Li
M L
ML
Wang
C Y
CY
Looney
D J
DJ
Wong-Staal
F
F
Arnold
E
E
Arnold
G F
GF
D000818:Animals; D002213:Capsid / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D036022:Capsid Proteins; D004279:DNA, Viral; D004220:Disulfides; D006168:Guinea Pigs; D015483:HIV Antibodies / Q000276:immunology; D015699:HIV Envelope Protein gp120 / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D015497:HIV-1 / Q000276:immunology*; D006367:HeLa Cells; D006801:Humans; D008958:Models, Molecular; D016296:Mutagenesis; D009500:Neutralization Tests; D010446:Peptide Fragments / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology; D012229:Rhinovirus / Q000235:genetics / Q000276:immunology*
D036022:Capsid Proteins; D004279:DNA, Viral; D004220:Disulfides; D015483:HIV Antibodies; D015699:HIV Envelope Protein gp120; C059762:HIV envelope protein gp120 (305-321); D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; C442544:VP2 protein, Rhinovirus