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Optimization of HIV-1 Envelope DNA Vaccine Candidates within Three Different Animal Models, Guinea Pigs, Rabbits and Cynomolgus Macaques.

Borggren M, Vinner L, Andresen BS, Grevstad B, Repits J, Melchers M, Elvang TL, Sanders RW, Martinon F, Dereuddre-Bosquet N, Bowles EJ, Stewart-Jones G, Biswas P, Scarlatti G, Jansson M, Heyndrickx L, Grand RL, Fomsgaard A - Vaccines (Basel) (2013)

Bottom Line: Codon-optimized genes, encoded secreted gp140 or membrane bound gp150, were modified for expression of stabilized soluble trimer gene products, and delivered individually or mixed.Specific IgG after repeated i.d. inoculations with electroporation confirmed in vivo expression and immunogenicity.It was concluded that a trivalent mix of non-modified gp140 genes from rationally selected clinical isolates was, in this study, the best option to induce high and broad NAb in the rabbit model, but this optimization does not directly translate into similar responses in cynomolgus macaques.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology Diagnostics and Virology, Statens Serum Institut, Copenhagen 2300, Denmark.

ABSTRACT
HIV-1 DNA vaccines have many advantageous features. Evaluation of HIV-1 vaccine candidates often starts in small animal models before macaque and human trials. Here, we selected and optimized DNA vaccine candidates through systematic testing in rabbits for the induction of broadly neutralizing antibodies (bNAb). We compared three different animal models: guinea pigs, rabbits and cynomolgus macaques. Envelope genes from the prototype isolate HIV-1 Bx08 and two elite neutralizers were included. Codon-optimized genes, encoded secreted gp140 or membrane bound gp150, were modified for expression of stabilized soluble trimer gene products, and delivered individually or mixed. Specific IgG after repeated i.d. inoculations with electroporation confirmed in vivo expression and immunogenicity. Evaluations of rabbits and guinea pigs displayed similar results. The superior DNA construct in rabbits was a trivalent mix of non-modified codon-optimized gp140 envelope genes. Despite NAb responses with some potency and breadth in guinea pigs and rabbits, the DNA vaccinated macaques displayed less bNAb activity. It was concluded that a trivalent mix of non-modified gp140 genes from rationally selected clinical isolates was, in this study, the best option to induce high and broad NAb in the rabbit model, but this optimization does not directly translate into similar responses in cynomolgus macaques.

No MeSH data available.


Related in: MedlinePlus

Immunization regimen and comparing antibody responses in syn.gp140Bx08 or syn.gp150Bx08. DNA vaccinated rabbits. (A) Schematic immunization schedule with vertical arrows indicating immunizations. Sera were collected before immunization (w0) and two weeks after last immunization (w14). (B) Average IgG response against recombinant gp120IIIb (rgp120IIIb) in immunized rabbits (n = 4). (C) Average neutralizing activity, expressed as IC50, of purified IgG from week 14 rabbit sera against pseudotype virus strains of clade B, C and A (SF162, Bx08, JR-FL, BaL, 92Br025 and 92RW009).
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vaccines-01-00305-f001: Immunization regimen and comparing antibody responses in syn.gp140Bx08 or syn.gp150Bx08. DNA vaccinated rabbits. (A) Schematic immunization schedule with vertical arrows indicating immunizations. Sera were collected before immunization (w0) and two weeks after last immunization (w14). (B) Average IgG response against recombinant gp120IIIb (rgp120IIIb) in immunized rabbits (n = 4). (C) Average neutralizing activity, expressed as IC50, of purified IgG from week 14 rabbit sera against pseudotype virus strains of clade B, C and A (SF162, Bx08, JR-FL, BaL, 92Br025 and 92RW009).

Mentions: Ten week old female iparous New Zealand white rabbits purchased from Charles River Laboratories were housed at Statens Serum Institute Animal Facility (Copenhagen, Denmark). Acclimatization was at least 10 days prior to any experimental procedures. Animal experiments were performed by certified animal handlers and according to the Animal Experimentation Act of Denmark and European Convention ETS 123. Groups of four rabbits were immunized at week 0 (three times during the first week), 4, 8 and 12 with 200 µg DNA injected intradermal (i.d.) and distributed at two injections sites (Figure 1A). The mode of “intensive” priming within week 0 (3 × 200 µg DNA) was initially compared with single DNA immunization and protein immunization. and demonstrated a more rapid and uniform antibody response than both other immunizations (Supplementary Figure 1). Subsequent electroporation using OncoVet™ system (CytoPulse Sciencies/Cellectis, Romainville, France) was done over each injected area. Four groups of rabbits were used, receiving syn.gp140Bx08, syn.gp150Bx08, syn.gp140mix (syn.gp140ctl21 + syn.gp140ctl27 + syn.gp140Bx08) or syn.gp140mix modified (syn.gp140Bx08 SOSIP.R6-IZ-H8 + syn.gp140ctl21 SOSIP.R6-IZ-H8 + syn.gp140ctl27 SOSIP.R6-IZ-H8). The amount of DNA constructs in the mixed formulations was 1:1:1, giving in total a 200 µg/immunization. In all four groups, blood was collected before immunization (w0) and two weeks after last immunization (w14). In addition, rabbits immunized with syn.gp140mix had blood collected at each re-immunization (w4, 8, 12) and rabbits immunized with syn.gp140mixSOSIP.R6-IZ-H8 had blood collected every week until w6 (w1, 2, 3, 4, 5, 6), then every second week until w14 (w8, 10, 12, 14).


Optimization of HIV-1 Envelope DNA Vaccine Candidates within Three Different Animal Models, Guinea Pigs, Rabbits and Cynomolgus Macaques.

Borggren M, Vinner L, Andresen BS, Grevstad B, Repits J, Melchers M, Elvang TL, Sanders RW, Martinon F, Dereuddre-Bosquet N, Bowles EJ, Stewart-Jones G, Biswas P, Scarlatti G, Jansson M, Heyndrickx L, Grand RL, Fomsgaard A - Vaccines (Basel) (2013)

Immunization regimen and comparing antibody responses in syn.gp140Bx08 or syn.gp150Bx08. DNA vaccinated rabbits. (A) Schematic immunization schedule with vertical arrows indicating immunizations. Sera were collected before immunization (w0) and two weeks after last immunization (w14). (B) Average IgG response against recombinant gp120IIIb (rgp120IIIb) in immunized rabbits (n = 4). (C) Average neutralizing activity, expressed as IC50, of purified IgG from week 14 rabbit sera against pseudotype virus strains of clade B, C and A (SF162, Bx08, JR-FL, BaL, 92Br025 and 92RW009).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4494233&req=5

vaccines-01-00305-f001: Immunization regimen and comparing antibody responses in syn.gp140Bx08 or syn.gp150Bx08. DNA vaccinated rabbits. (A) Schematic immunization schedule with vertical arrows indicating immunizations. Sera were collected before immunization (w0) and two weeks after last immunization (w14). (B) Average IgG response against recombinant gp120IIIb (rgp120IIIb) in immunized rabbits (n = 4). (C) Average neutralizing activity, expressed as IC50, of purified IgG from week 14 rabbit sera against pseudotype virus strains of clade B, C and A (SF162, Bx08, JR-FL, BaL, 92Br025 and 92RW009).
Mentions: Ten week old female iparous New Zealand white rabbits purchased from Charles River Laboratories were housed at Statens Serum Institute Animal Facility (Copenhagen, Denmark). Acclimatization was at least 10 days prior to any experimental procedures. Animal experiments were performed by certified animal handlers and according to the Animal Experimentation Act of Denmark and European Convention ETS 123. Groups of four rabbits were immunized at week 0 (three times during the first week), 4, 8 and 12 with 200 µg DNA injected intradermal (i.d.) and distributed at two injections sites (Figure 1A). The mode of “intensive” priming within week 0 (3 × 200 µg DNA) was initially compared with single DNA immunization and protein immunization. and demonstrated a more rapid and uniform antibody response than both other immunizations (Supplementary Figure 1). Subsequent electroporation using OncoVet™ system (CytoPulse Sciencies/Cellectis, Romainville, France) was done over each injected area. Four groups of rabbits were used, receiving syn.gp140Bx08, syn.gp150Bx08, syn.gp140mix (syn.gp140ctl21 + syn.gp140ctl27 + syn.gp140Bx08) or syn.gp140mix modified (syn.gp140Bx08 SOSIP.R6-IZ-H8 + syn.gp140ctl21 SOSIP.R6-IZ-H8 + syn.gp140ctl27 SOSIP.R6-IZ-H8). The amount of DNA constructs in the mixed formulations was 1:1:1, giving in total a 200 µg/immunization. In all four groups, blood was collected before immunization (w0) and two weeks after last immunization (w14). In addition, rabbits immunized with syn.gp140mix had blood collected at each re-immunization (w4, 8, 12) and rabbits immunized with syn.gp140mixSOSIP.R6-IZ-H8 had blood collected every week until w6 (w1, 2, 3, 4, 5, 6), then every second week until w14 (w8, 10, 12, 14).

Bottom Line: Codon-optimized genes, encoded secreted gp140 or membrane bound gp150, were modified for expression of stabilized soluble trimer gene products, and delivered individually or mixed.Specific IgG after repeated i.d. inoculations with electroporation confirmed in vivo expression and immunogenicity.It was concluded that a trivalent mix of non-modified gp140 genes from rationally selected clinical isolates was, in this study, the best option to induce high and broad NAb in the rabbit model, but this optimization does not directly translate into similar responses in cynomolgus macaques.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology Diagnostics and Virology, Statens Serum Institut, Copenhagen 2300, Denmark.

ABSTRACT
HIV-1 DNA vaccines have many advantageous features. Evaluation of HIV-1 vaccine candidates often starts in small animal models before macaque and human trials. Here, we selected and optimized DNA vaccine candidates through systematic testing in rabbits for the induction of broadly neutralizing antibodies (bNAb). We compared three different animal models: guinea pigs, rabbits and cynomolgus macaques. Envelope genes from the prototype isolate HIV-1 Bx08 and two elite neutralizers were included. Codon-optimized genes, encoded secreted gp140 or membrane bound gp150, were modified for expression of stabilized soluble trimer gene products, and delivered individually or mixed. Specific IgG after repeated i.d. inoculations with electroporation confirmed in vivo expression and immunogenicity. Evaluations of rabbits and guinea pigs displayed similar results. The superior DNA construct in rabbits was a trivalent mix of non-modified codon-optimized gp140 envelope genes. Despite NAb responses with some potency and breadth in guinea pigs and rabbits, the DNA vaccinated macaques displayed less bNAb activity. It was concluded that a trivalent mix of non-modified gp140 genes from rationally selected clinical isolates was, in this study, the best option to induce high and broad NAb in the rabbit model, but this optimization does not directly translate into similar responses in cynomolgus macaques.

No MeSH data available.


Related in: MedlinePlus