The APL 400-047 DNA plasmid vaccine expresses the human immunodeficiency virus type-1 core structural proteins, and reverse transcriptase and integrase proteins. Both the reverse transcriptase and integrase proteins are partially deleted and non-functional. These genes are derived from the HIV-1 strain HXB2 and are inserted into the GENEVAX^® DNA plasmid backbone. APL 400-047 is formulated with the facilitating agent bupivacaine, an amide-type local anesthetic agent.

HIVAC-1e: The vaccinia virus v-envNY5 construct contains the entire coding sequence of the envelope glycoproteins of HIV strain LAV-1. The vaccinia viral vector used was a derivative of the New York Board of Health strain obtained by plaque purification from a commercial smallpox vaccine (DryVax, Lot #321501G, Wyeth Laboratories) using three successive plaque purifications in tissue culture cells. The master seed lot of the recombinant virus is prepared in human diploid MRC-5 cells.

HIV p17/p24:Ty-VLP is a particulate immunogen produced in yeast. The expression plasmid encoding the p1/p17/p24 fusion gene is introduced into yeast cells. The cells are grown to high cell density in a medium containing glucose. Expression of the fusion protein is then induced by adding galactose which switches on the production via a cascading promoter system.

ENV 2-3 is a non-glycosylated, denatured polypeptide containing the entire primary sequence of the gp120 envelope protein of HIV-1. It is obtained from the SF-2 isolate of HIV and produced in genetically engineered yeast.

The HIV SF-2 gp120 candidate vaccine is a recombinant product produced in genetically engineered CHO cells under the control of the cytomegalovirus (hCMV) immediate early - 1 (IE-1) promoter. The antigen is formulated in phosphate buffered saline. The most commonly used adjuvant is MF59, a microfluidized oil-in-water emulsion.

Recombinant subunit p24 antigen is derived from the SF-2 p24 isolate of HIV-1 and is expressed in genetically engineered Saccharomyces cerevisiae organisms and formulated in combination with MF59 adjuvant emulsion.

032

The immunogen IIIB rgp120-HIV-1 produced in a genetically engineered mammalian cell (CHO) adsorbed to aluminum hydroxide adjuvant with 0.1 mg thimerosal.

Recombinant MN rgp120/HIV-1 is a protein produced by genetically engineered mammalian cell lines (CHO) and isolated in a highly purified form. MN rgp120/HIV-1 is adsorbed to aluminum hydroxide adjuvant with 0.1 mg thimerosal added as a preservative.

Recombinant soluble gp120/HIV-1 (referred to as rsgp120) is the same glycoprotein, as described above, without alum. Alum and/or QS21 adjuvant are added to the rsgp120 protein to make the various dosing solutions for studies that use rsgp120.

AIDSVAX^TM B/E is a highly purified mixture of recombinant MN rgp120 (subtype B) and A244 rgp120 (subtype E). These gp120 proteins are produced by genetically engineered mammalian cell lines (CHO). AIDSVAX^TM B/E is adsorbed onto an aluminum hydroxide gel adjuvant.

rgp160 IIIB. Vaccinia-derived HIV-1 IIIB recombinant envelope glycoprotein (IIIB rgp160) is obtained using a recombinant vaccinia virus/bacteriophage T7 hybrid system, to express the HIV env gene in mammalian cells. Briefly, one vaccinia recombinant contains the bacteriophage T7 polymerase gene under the direction of the vaccinia virus P7.5 promoter. This is infected together into vero cells with another vaccinia recombinant containing the HIV env gene under the direction of a T7 promoter. The rgp160 is purified from infected vero cells following extraction, lentil-lectin chromatography and immune affinity chromatography. The adjuvant used is alum + deoxycholate (DOC).

rgp160 MN. Vaccinia derived HIV-1 MN recombinant envelope glycoprotein (rgp160) is obtained by using a recombinant vaccinia virus/bacteriophage T7 hybrid system to express the HIV env gene in mammalian cells. Briefly, one vaccinia recombinant contains the bacteriophage T7 polymerase gene under the direction of the vaccinia virus P7.5 promoter. Vero cells are co-infected with this recombinant and another vaccinia recombinant containing the HIV gp160 env gene under the direction of a T7 promoter. The rgp160 is purified from infected Vero cells following extraction, lentil-lectin chromatography and immune affinity chromatography. The adjuvant used is alum + deoxycholate (DOC).

VaxSyn: VaxSyn, HIV-1 (Human T-Lymphotrophic Virus Type III recombinant gp160 experimental vaccine, aluminum phosphate adsorbed) is a non-infectious subunit vaccine containing the HIV-1 gp160 envelope protein (LAV strain). The gp160 contained in the vaccine is at least 95% pure. Because this protein is prepared from recombinant baculoviruses and insect cell cultures, it is free of association with human blood or blood products. The gp160 protein is adsorbed onto an aluminum phosphate adjuvant.

The recombinant canarypox-HIV-1 MN gp160 virus, ALVAC vCP125, was produced by homologous recombination between the plasmid pC5HIVMNE and the genomic DNA of ALVAC (canarypox), thereby substituting the gp160 gene for the open reading frame C5. The gp160 gene is under the control of the vaccinia H6 promoter. Plaques were selected by hybridization with radioactive DNA gp160 probes; after several cycles of purification, one plaque was selected. Viral stocks are grown in chick embryo fibroblasts. The DNA from this viral stock has been analyzed by restriction enzyme analyses; the pattern corresponds exactly to that predicted. The sequence of the MN gp160 gene in vCP125 is identical to the gp160 gene in the plasmid pC5HIVMNE. Expression of the protein gp160 by cells infected with vCP125 has been confirmed by immunofluorescence. By SDS-PAGE, Vero cells infected with ALVAC-gp160 have been shown to produce radioactively labeled gp160, gp120, and gp41. ALVAC vCP125 does not appear to replicate in mammalian cells; titers of ALVAC recovered from 6 mammalian cell lines after 72 hours incubation were lower than the original virus inoculum. After penetration into a mammalian cell, the virion is uncoated and the recombinant gene is expressed.

The recombinant canarypox-HIV-1 virus, ALVAC vCP205, is a second generation vaccine that can be used to induce a humoral and cellular response against several antigens. It expresses these products of HIV-1 genes:

• the gag gene expressing the gag p55-poly protein of the HIV-1 LAI strain
• the protease gene, expressing the p15 protein of the HIV-1 LAI strain
• a part of the env gene expressing the glycoprotein (gp) 120 of the HIV-1 MN strain, and the anchoring region of gp41, the transmembrane glycoprotein (TM) of the HIV-1 LAI strain.

The gag gene, which codes for virus core antigens, is a well-conserved gene. It codes for a polyprotein (55kDa) which matures into the core proteins, p24, p17 and p15, the last of which is then split into p9 and p6. This maturation process is catalyzed by the HIV protease coded by a part of the pol gene. The HIV-1 gag proteins synthesized in cells by various expression systems form virus-like particles before leaving the cells. The co-expression of env components in the same cell as gag proteins results in a gag-env interaction, followed by the enveloping of the gag particles. The viral particles bud from the cell membrane and are similar to HIV virions in their morphology and protein content.

The recombinant canarypox-HIV-1 virus, ALVAC vCP300, is a second generation vaccine that can be used to induce a humoral and cellular response against several antigens. It expresses these products of HIV-1 genes:

• the gag gene expressing the gag p55-poly protein of the HIV-1 LAI strain
• the protease gene, expressing the p15 protein of the HIV-1 LAI strain
• a part of the env gene expressing the glycoprotein (gp) 120 of the HIV-1 MN strain, and the anchoring region of gp41, the transmembrane glycoprotein (TM) of the HIV-1 LAI strain
• a part of the HIV-1 nef gene (LAI strain) gene encoding two peptides (9.1 and 3.3 kDa) containing multiple CTL epitopes
• a part of the HIV-1 pol gene (LAI strain) encoding three peptides (5.5, 7.2 and 7.0 kDa) containing multiple CTL epitopes

The production of non-infectious virus-like particles is a feature of this vaccine. This new vaccine should make it possible to extend the range of immune responses, to express gp120 at the surface of the infected cells, and to induce the in vivo production of structures similar to viruses (virus-like particles or pseudovirions) by the co-expression of an envelope and gag component. The deletion of part of the sequences of the envelope transmembrane region (gp41) makes it easier to distinguish between HIV-infected subjects and vaccinated subjects since most HIV-infected subjects show antibodies directed against products of the particular gp41 sequences deleted in vCP300.

Pasteur-Merieux/

Connaught

ALVAC-HIV (vCP1433) vaccine is a preparation of a modified recombinant canarypox virus expressing the products of the HIV-1 env, gag and the potion of the pol gene encoding the protease genes, and a synthetic polypeptide encompassing the known human CTL epitopes from the nef and pol gene products.

Recombinant ALVAC-MN120TMGNPst (vCP1433) was generated by insertion of an expression cassette encoding a synthetic polypeptide containing several of the known human pol CTL epitopes and all of the known human nef CTL epitopes into vCP205. ALVAC-HIV (vCP1433) was grown in chick embryo fibroblasts (CEF) derived from pathogen-free chicken eggs (SPF).

ALVAC-HIV (vCP1452) vaccine is a preparation of a modified recombinant canarypox virus expressing the products of the HIV-1 env, gag and the pol gene encoding the protease, and a synthetic polypeptide encompassing several known human CTL epitopes from the nef and pol gene products. Recombinant ALVAC-MN120TMGNPst (vCP1452) was generated by insertion of the vector modifying sequences encoding E3L and K3L into the C6 site of recombinant vCP1433.

Recombinant protein gp160MN/LAI-2 is an envelope glycoprotein from HIV-1 virus expressed by VV.TG.9150 vaccinia virus on BHK21 cells. This recombinant virus is a derivative of VV.TG.1163 where the HIV-1 LAI gp120 coding sequence has been replaced by the HIV-1 MN counterpart. Env hybrid HIV-1 MN/LAI glycoprotein gene is inserted under the control of the vaccinia H5R promotor into the TK locus of the Copenhagen vaccinia virus. Recombinant gp160MN/LAI-2 was produced by VV.TG.9150 infected BHK21 cells, followed by a multistep purification process.

TBC-3B is a live recombinant vaccinia-HIV-1 virus that expresses multiple genes of HIV-1. The parental strain used for the generation of this vaccine was plaque purified from the licensed Wyeth (New York City Board of Health) vaccine strain of vaccinia virus. The insertion of the HIV-1 (strain IIIB) env, gag and pol genes, each under the transcriptional direction of a vaccinia promoter element, into the genome of this parental virus has resulted in the generation of a multivalent recombinant virus that efficiently expresses fully processed, authentically configured HIV-1 polypeptides encoded by those genes. Upon administration to animals, this vaccine presents multiple conserved and variable HIV-1 epitopes to the immune system, and elicits both humoral and cell mediated immune responses to the expressed polypeptides.

TBC-3B was originally designed for administration by scarification. The estimated human dose delivered by this route is ~1.0 µl or 10⁶ pfu of virus. TBC-3B may also be administered by intradermal or subcutaneous injection. A volume of 100 µl, which is practical for human use by these routes, will deliver a maximum dose of ~10⁸ pfu of virus.

TBC-3B lot #3-101191: virus was grown in RK₁₃ (rabbit kidney) cells and purified through a 36% sucrose cushion. Following virus titration using BSC-40 cells, the concentration of the bulk material was adjusted by the addition of PBS with 10% glycerol.

TBC-3B Lot #6-082195: The virus was grown in CV-1 (African green monkey kidney) cells and purified through a 36% sucrose cushion. Following virus titration using BSC-40 cells, the concentration of the bulk material was adjusted by the addition of PBS with 10% glycerol. The final titer of Lot #6-082195 is 3.67x10⁹ plaque forming units (pfu) per ml.

SynVac HIV-1 MN octameric V3 or PND immunogen is a synthetic peptide prototype vaccine based on eight V3-derived homologous peptides attached to a heptalysyl core to form radially branched structures and formulated in alum.

Multivalent HIV-1 V3 peptide immunogen is a mixture of HIV-1 gp120 principal neutralizing domain (PND), V3 octameric synthetic peptides from 15 HIV-1 subtypes and formulated in alum. Each component of the vaccine consists of eight V3-derived homologous peptides attached to a heptalysyl core to form radially branched structures.

Microparticulate Monovalent HIV-1 MN V3 octameric peptide vaccine is a branched peptide immunogen consisting of eight V3-derived homologous peptides attached to a heptalysyl core to form radially branched structures, and entrapped in microparticles polylactide co-glycosides and polylactide polymers.

HIV gag lipopeptide, P3C541b, is a linear peptide representing a sequence of the HIV gag protein conjugated to a lipid moiety and solubilized for parenteral administration.

Attenuated S. Typhi CVD 908 is a live vector expressing an engineered prokaryotic expression cassette that encodes HIV-1 gp120 or truncated gp120, resulting in secretion of rgp120 into the cytoplasm or periplasmic compartment of the S. Typhi vector

HIV-1 gp120 C4-V3 is a hybrid polyvalent peptide immunogen in mineral oil + mannose mono-oleate (IFA). Four synthetic peptides based on the HIV-1 clade B strains MN, EV91, RF and CANO are included in the candidate vaccine