Retroviral vectors for vaccine development: induction of HIV-1-specific humoral and cellular immune responses in rhesus macaques using a novel MLV(HIV-1) pseudotype vector.
Retroviral vectors have yet not been tested for their potential as vaccines despite their frequent utilization in gene therapy allowing for highly efficient gene transfer into a number of cell types and their suitability for large-scale production in biotechnology. To investigate MLV-based vectors suitability for inducing immune response against HIV-1-antigens, we generated a MLV(HIV-1) pseudotype vector enabling CD4-specific transduction of HIV-1 genes env, vpu, tat and rev originating from the pathogenic SHIV-89.6P. Functional expression of the lentiviral genes in packaging cells, human and rhesus CD4+ target cells was demonstrated by various assays. Following highly efficient ex vivo transduction, up to 3.4x10(7) autologous, transfer vector-positive rhesus peripheral blood mononuclear cells (rhPBMCs) were re-inoculated into a rhesus macaque. Five weeks after the initial inoculation HIV-1 Env-specific antibodies were detected using ELISA. ELIspot-assay revealed the induction of a HIV-1 Rev and Env-specific CTL-response 7.5 weeks after immunization. Thus, these novel MLV(HIV-1) vectors facilitate efficient transduction and subsequent expression of HIV-1-genes in CD4-positive host cells. Induction of both humoral and cellular HIV-1-specific immune responses in vivo confirmed their potential as an effective HIV-1 vaccine to be further studied in SHIV/rhesus macaque model of lentivirus infection.
Paul-Ehrlich-Institute, Department of Medical Biotechnology, Paul-Ehrlich-Strasse 51-59, D-63225 Langen, Germany.
Jeanette
J
Neumann
Jörn
J
Stitz
Renate
R
König
Eric
E
Seibold
Stephen
S
Norley
Egbert
E
Flory
Klaus
K
Cichutek
D016915:AIDS Vaccines / Q000008:administration & dosage* / Q000235:genetics; D000818:Animals; D000917:Antibody Formation / Q000276:immunology*; D003001:Cloning, Molecular / Q000379:methods; D005822:Genetic Vectors / Q000235:genetics; D015497:HIV-1 / Q000235:genetics* / Q000276:immunology*; D000949:Histocompatibility Antigens Class II / Q000276:immunology*; D008253:Macaca mulatta; D012190:Retroviridae / Q000235:genetics*; D014162:Transfection / Q000379:methods*
D016915:AIDS Vaccines; D000949:Histocompatibility Antigens Class II