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Comparative Study
. 2007 Jun 21;25(26):4949-59.
doi: 10.1016/j.vaccine.2007.01.118. Epub 2007 Feb 26.

Comparison of immunogenicity between codon optimized HIV-1 Thailand subtype B gp140 and gp145 vaccines

Yanmin Wan  1 , Lan Wu, Lianxing Liu, Jianqing Xu, Ying Liu, Yong Liu, Yiming Shao
Affiliations
Comparative Study

Comparison of immunogenicity between codon optimized HIV-1 Thailand subtype B gp140 and gp145 vaccines

Yanmin Wan et al. Vaccine. .

Abstract

HIV-1 pandemic posed an unprecedented challenge to the global health and it is believed that an effective vaccine will be the final solution against HIV-1. HIV-1 envelope is the primary immunogen in developing neutralization antibody based HIV vaccine. To define the suitable Env derived immunogen, we systemically compared the immunogenicity of gp140 and gp145 in a DNA vaccination alone and a prime-boost modalities. Two DNA vaccines and two recombinant Tiantan vaccinia vaccines (rTTV) were constructed for vaccination of female Balb/c mice. Elispot assay was used to read out the T cell immunity and ELISA assay was used to quantify antibody immunity. PLL (poly-L-leucine)-ELISA assay was used in linear antibody epitope mapping. Mice primed with gp145 tended to elicit more Env-specific T cells responses than those primed with gp140, significant difference was observed in DNA immunization alone. The ultimate T cell responses in prime-boost regimen tend to be determined mainly by the priming efficacy. Linear antibody epitope mapping displayed that sera raised by gp145 priming were vigorously reactive to more peptides than that by gp140. Our data demonstrated HIV-1 Thailand B-derived gp145 may raise higher T-cell responses and broader linear peptide-specific antibody responses than gp140 does. However, it remains to be determined that how these observations are relevant to the neutralization of antibody activities.

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Figures

Figure 1
Figure 1
In vitro expressions of DNA vaccines encoding HIV-1rl42 gp140 and gp145 in Hela cells (upper panel) and of rTTVs in primary chicken embryo cells (bottom panel) were assayed by Western Blotting of cell lysate. Blank vector pSV1.0 and wild type Tiantan vaccinia virus was used as mock controls. Transfection and infection were carried out separately for different vaccines, cell numbers used for preparation of cell lysate were varied.
Figure 2
Figure 2
Env-specific T cell immune responses among different groups. Mice immunized with different regimens of vaccines (as scheduled in table 1) were sacrificed and splenocytes were harvested. Four peptide pools derived from consensus B clade Env were used as stimuli in Elispot assay, T-cell responses for each mouse were compiled from 4 peptide pool stimulations and then grouped in the figure. Positive Elispot control (stimulated with PMA and Inomycin) and negative control (stimulated with no peptides) were also included. Peptide pool specific IFN-γ Elispot responses were considered as positive only when the responses were 4-fold above negative control with no peptide stimulation. Statistical significances were only observed between gp140 and other 5 groups. Control groups immunized with mock vectors raise no T cell immune responses.
Figure 3
Figure 3
Gp120-specific binding antibodies among different groups. Mice immunized with different regimens of vaccines (as scheduled in table 1) were sacrificed and sera were collected. Gp120 expressed in E.coli was used in ELISA assay and endpoint titers were determined by the last dilution whose OD was >2 folds than that at the corresponding dilution of mice sera immunized with mock control group (sv1.0/wild type TTV). Control groups immunized with mock vectors raise no humoral responses. No significant differences were observed among all 6 groups.
Figure 4
Figure 4
Results of linear antibody epitope mapping. For each group, the sera from mice in the same group were combined together in equal volume and mixed. The mixed serum was then assayed in duplicated wells. The averages of duplicated original OD values from vaccination groups were graphed into figures. The straight-scatter lines in the figures indicate the threshold value, which is 2-fold of the OD values from mock vector controls against each corresponding peptide(2-fold of sv1.0 is the cutoff value for groups immunized with DNA only; 2-fold of sv1.0+wild type TTV is the cutoff value for groups immunized DNA plus rTTV). Though vaccination stimulated no significant responses to the majority of peptides, several highly reactive epitopes were identified across the full amino acid sequences of HIV-1rl42 gp145, including p22~p27 of C1, P76/p77 of V3, p86/p87 of C3, p116~p119 of C5 and p145/146 between the 2 HR (heptad repeat) regions.
Figure 4
Figure 4
Results of linear antibody epitope mapping. For each group, the sera from mice in the same group were combined together in equal volume and mixed. The mixed serum was then assayed in duplicated wells. The averages of duplicated original OD values from vaccination groups were graphed into figures. The straight-scatter lines in the figures indicate the threshold value, which is 2-fold of the OD values from mock vector controls against each corresponding peptide(2-fold of sv1.0 is the cutoff value for groups immunized with DNA only; 2-fold of sv1.0+wild type TTV is the cutoff value for groups immunized DNA plus rTTV). Though vaccination stimulated no significant responses to the majority of peptides, several highly reactive epitopes were identified across the full amino acid sequences of HIV-1rl42 gp145, including p22~p27 of C1, P76/p77 of V3, p86/p87 of C3, p116~p119 of C5 and p145/146 between the 2 HR (heptad repeat) regions.
Figure 4
Figure 4
Results of linear antibody epitope mapping. For each group, the sera from mice in the same group were combined together in equal volume and mixed. The mixed serum was then assayed in duplicated wells. The averages of duplicated original OD values from vaccination groups were graphed into figures. The straight-scatter lines in the figures indicate the threshold value, which is 2-fold of the OD values from mock vector controls against each corresponding peptide(2-fold of sv1.0 is the cutoff value for groups immunized with DNA only; 2-fold of sv1.0+wild type TTV is the cutoff value for groups immunized DNA plus rTTV). Though vaccination stimulated no significant responses to the majority of peptides, several highly reactive epitopes were identified across the full amino acid sequences of HIV-1rl42 gp145, including p22~p27 of C1, P76/p77 of V3, p86/p87 of C3, p116~p119 of C5 and p145/146 between the 2 HR (heptad repeat) regions.
Figure 4
Figure 4
Results of linear antibody epitope mapping. For each group, the sera from mice in the same group were combined together in equal volume and mixed. The mixed serum was then assayed in duplicated wells. The averages of duplicated original OD values from vaccination groups were graphed into figures. The straight-scatter lines in the figures indicate the threshold value, which is 2-fold of the OD values from mock vector controls against each corresponding peptide(2-fold of sv1.0 is the cutoff value for groups immunized with DNA only; 2-fold of sv1.0+wild type TTV is the cutoff value for groups immunized DNA plus rTTV). Though vaccination stimulated no significant responses to the majority of peptides, several highly reactive epitopes were identified across the full amino acid sequences of HIV-1rl42 gp145, including p22~p27 of C1, P76/p77 of V3, p86/p87 of C3, p116~p119 of C5 and p145/146 between the 2 HR (heptad repeat) regions.
Figure 4
Figure 4
Results of linear antibody epitope mapping. For each group, the sera from mice in the same group were combined together in equal volume and mixed. The mixed serum was then assayed in duplicated wells. The averages of duplicated original OD values from vaccination groups were graphed into figures. The straight-scatter lines in the figures indicate the threshold value, which is 2-fold of the OD values from mock vector controls against each corresponding peptide(2-fold of sv1.0 is the cutoff value for groups immunized with DNA only; 2-fold of sv1.0+wild type TTV is the cutoff value for groups immunized DNA plus rTTV). Though vaccination stimulated no significant responses to the majority of peptides, several highly reactive epitopes were identified across the full amino acid sequences of HIV-1rl42 gp145, including p22~p27 of C1, P76/p77 of V3, p86/p87 of C3, p116~p119 of C5 and p145/146 between the 2 HR (heptad repeat) regions.
Figure 4
Figure 4
Results of linear antibody epitope mapping. For each group, the sera from mice in the same group were combined together in equal volume and mixed. The mixed serum was then assayed in duplicated wells. The averages of duplicated original OD values from vaccination groups were graphed into figures. The straight-scatter lines in the figures indicate the threshold value, which is 2-fold of the OD values from mock vector controls against each corresponding peptide(2-fold of sv1.0 is the cutoff value for groups immunized with DNA only; 2-fold of sv1.0+wild type TTV is the cutoff value for groups immunized DNA plus rTTV). Though vaccination stimulated no significant responses to the majority of peptides, several highly reactive epitopes were identified across the full amino acid sequences of HIV-1rl42 gp145, including p22~p27 of C1, P76/p77 of V3, p86/p87 of C3, p116~p119 of C5 and p145/146 between the 2 HR (heptad repeat) regions.
Figure 4
Figure 4
Results of linear antibody epitope mapping. For each group, the sera from mice in the same group were combined together in equal volume and mixed. The mixed serum was then assayed in duplicated wells. The averages of duplicated original OD values from vaccination groups were graphed into figures. The straight-scatter lines in the figures indicate the threshold value, which is 2-fold of the OD values from mock vector controls against each corresponding peptide(2-fold of sv1.0 is the cutoff value for groups immunized with DNA only; 2-fold of sv1.0+wild type TTV is the cutoff value for groups immunized DNA plus rTTV). Though vaccination stimulated no significant responses to the majority of peptides, several highly reactive epitopes were identified across the full amino acid sequences of HIV-1rl42 gp145, including p22~p27 of C1, P76/p77 of V3, p86/p87 of C3, p116~p119 of C5 and p145/146 between the 2 HR (heptad repeat) regions.
Figure 4
Figure 4
Results of linear antibody epitope mapping. For each group, the sera from mice in the same group were combined together in equal volume and mixed. The mixed serum was then assayed in duplicated wells. The averages of duplicated original OD values from vaccination groups were graphed into figures. The straight-scatter lines in the figures indicate the threshold value, which is 2-fold of the OD values from mock vector controls against each corresponding peptide(2-fold of sv1.0 is the cutoff value for groups immunized with DNA only; 2-fold of sv1.0+wild type TTV is the cutoff value for groups immunized DNA plus rTTV). Though vaccination stimulated no significant responses to the majority of peptides, several highly reactive epitopes were identified across the full amino acid sequences of HIV-1rl42 gp145, including p22~p27 of C1, P76/p77 of V3, p86/p87 of C3, p116~p119 of C5 and p145/146 between the 2 HR (heptad repeat) regions.
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