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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: Specific IgG after repeated i.d. inoculations with electroporation confirmed in vivo expression and immunogenicity.The superior DNA construct in rabbits was a trivalent mix of non-modified codon-optimized gp140 envelope genes.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.


Schematic representation of HIV-1 envelope DNA constructs and protein expression. DNA constructs encoding gp140. The tissue plasminogen-activator leader sequence (tPA) and the region encoding the variable regions V1 to V5 are indicated (grey boxes). (A) The gp140ctl21/27 construct with V1-V5 region from ctl21 and ctl27 env flanked by Bx08 env. (B) DNA construct encoding modified gp140 including the SOSIP amino acid substitutions A501C, T605C and I559P (SOSIP), the hexa-arginine cleavage site (R6), the introduced isoleucine-zipper motif (IZ) and the histidine tag (H8). (C) Western blot analysis of protein expression (SDS-PAGE) and oligomerization (Blue-Native PAGE) of EnvBx08 constructs, encoding gp120, gp140, gp140SOSIP.R6 and gp140SOSIP.R6-IZ-H8.
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vaccines-01-00305-f003: Schematic representation of HIV-1 envelope DNA constructs and protein expression. DNA constructs encoding gp140. The tissue plasminogen-activator leader sequence (tPA) and the region encoding the variable regions V1 to V5 are indicated (grey boxes). (A) The gp140ctl21/27 construct with V1-V5 region from ctl21 and ctl27 env flanked by Bx08 env. (B) DNA construct encoding modified gp140 including the SOSIP amino acid substitutions A501C, T605C and I559P (SOSIP), the hexa-arginine cleavage site (R6), the introduced isoleucine-zipper motif (IZ) and the histidine tag (H8). (C) Western blot analysis of protein expression (SDS-PAGE) and oligomerization (Blue-Native PAGE) of EnvBx08 constructs, encoding gp120, gp140, gp140SOSIP.R6 and gp140SOSIP.R6-IZ-H8.

Mentions: Plasma samples (n = 35) from Danish HIV-1-infected treatment-naïve individuals were collected [46] and screened for neutralization against HIV-1 virus isolates, four clade B and one A1D intersubtype recombinant [47] (Table 1). As expected, the sensitivity to neutralization varied among the virus isolates, with clade B HIV-1BaL being most sensitive to neutralization and recombinant A1D HIV-1DK1 least sensitive. In many samples, neutralization was primarily directed against one or two viruses, but in 17 sera (49%) the neutralizing effect was detected against all five isolates, including the A1D recombinant. Among these, two plasma samples, ctl21 and ctl27, obtained from a male and a female with 9 and 3.5 years of infection, respectively, displayed robust and balanced neutralization titers against all five viruses. To test the hypothesis, the env region including V1-V5 of the clade B virus isolates from ctl21 and ctl27 were cloned, sequenced and synthesized as codon-optimized DNA vaccine constructs, flanked by the N- and C-terminal region of gp120 and the extracellular part of the gp41 region from the HIV-1Bx08env cassette (see Figure 3A and [18]). The constructs were control sequenced and tested for successful in vitro expression of functional envelope glycoproteins (CD4 binding) (data not shown).


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)

Schematic representation of HIV-1 envelope DNA constructs and protein expression. DNA constructs encoding gp140. The tissue plasminogen-activator leader sequence (tPA) and the region encoding the variable regions V1 to V5 are indicated (grey boxes). (A) The gp140ctl21/27 construct with V1-V5 region from ctl21 and ctl27 env flanked by Bx08 env. (B) DNA construct encoding modified gp140 including the SOSIP amino acid substitutions A501C, T605C and I559P (SOSIP), the hexa-arginine cleavage site (R6), the introduced isoleucine-zipper motif (IZ) and the histidine tag (H8). (C) Western blot analysis of protein expression (SDS-PAGE) and oligomerization (Blue-Native PAGE) of EnvBx08 constructs, encoding gp120, gp140, gp140SOSIP.R6 and gp140SOSIP.R6-IZ-H8.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

vaccines-01-00305-f003: Schematic representation of HIV-1 envelope DNA constructs and protein expression. DNA constructs encoding gp140. The tissue plasminogen-activator leader sequence (tPA) and the region encoding the variable regions V1 to V5 are indicated (grey boxes). (A) The gp140ctl21/27 construct with V1-V5 region from ctl21 and ctl27 env flanked by Bx08 env. (B) DNA construct encoding modified gp140 including the SOSIP amino acid substitutions A501C, T605C and I559P (SOSIP), the hexa-arginine cleavage site (R6), the introduced isoleucine-zipper motif (IZ) and the histidine tag (H8). (C) Western blot analysis of protein expression (SDS-PAGE) and oligomerization (Blue-Native PAGE) of EnvBx08 constructs, encoding gp120, gp140, gp140SOSIP.R6 and gp140SOSIP.R6-IZ-H8.
Mentions: Plasma samples (n = 35) from Danish HIV-1-infected treatment-naïve individuals were collected [46] and screened for neutralization against HIV-1 virus isolates, four clade B and one A1D intersubtype recombinant [47] (Table 1). As expected, the sensitivity to neutralization varied among the virus isolates, with clade B HIV-1BaL being most sensitive to neutralization and recombinant A1D HIV-1DK1 least sensitive. In many samples, neutralization was primarily directed against one or two viruses, but in 17 sera (49%) the neutralizing effect was detected against all five isolates, including the A1D recombinant. Among these, two plasma samples, ctl21 and ctl27, obtained from a male and a female with 9 and 3.5 years of infection, respectively, displayed robust and balanced neutralization titers against all five viruses. To test the hypothesis, the env region including V1-V5 of the clade B virus isolates from ctl21 and ctl27 were cloned, sequenced and synthesized as codon-optimized DNA vaccine constructs, flanked by the N- and C-terminal region of gp120 and the extracellular part of the gp41 region from the HIV-1Bx08env cassette (see Figure 3A and [18]). The constructs were control sequenced and tested for successful in vitro expression of functional envelope glycoproteins (CD4 binding) (data not shown).

Bottom Line: Specific IgG after repeated i.d. inoculations with electroporation confirmed in vivo expression and immunogenicity.The superior DNA construct in rabbits was a trivalent mix of non-modified codon-optimized gp140 envelope genes.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.