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Establishing the pig as a large animal model for vaccine development against human cancer.

Overgaard NH, Frøsig TM, Welner S, Rasmussen M, Ilsøe M, Sørensen MR, Andersen MH, Buus S, Jungersen G - Front Genet (2015)

Bottom Line: Taking advantage of recombinant swine MHC class I molecules (SLAs), the peptide-SLA complex stability was measured for 198 IDO- or RhoC-derived 9-11mer peptides predicted to bind to SLA-1(*)04:01, -1(*)07:02, -2(*)04:01, -2(*)05:02, and/or -3(*)04:01.This identified 89 stable (t½ ≥ 0.5 h) peptide-SLA complexes.By IFN-γ release in PBMC cultures we monitored the vaccine-induced peptide-specific CTL responses, and found responses to both IDO- and RhoC-derived peptides across all groups with no adjuvant being superior.

View Article: PubMed Central - PubMed

Affiliation: Department of Immunology and Vaccinology, National Veterinary Institute, Technical University of Denmark Copenhagen, Denmark.

ABSTRACT
Immunotherapy has increased overall survival of metastatic cancer patients, and cancer antigens are promising vaccine targets. To fulfill the promise, appropriate tailoring of the vaccine formulations to mount in vivo cytotoxic T cell (CTL) responses toward co-delivered cancer antigens is essential. Previous development of therapeutic cancer vaccines has largely been based on studies in mice, and the majority of these candidate vaccines failed to induce therapeutic responses in the subsequent human clinical trials. Given that antigen dose and vaccine volume in pigs are translatable to humans and the porcine immunome is closer related to the human counterpart, we here introduce pigs as a supplementary large animal model for human cancer vaccine development. IDO and RhoC, both important in human cancer development and progression, were used as vaccine targets and 12 pigs were immunized with overlapping 20mer peptides spanning the entire porcine IDO and RhoC sequences formulated in CTL-inducing adjuvants: CAF09, CASAC, Montanide ISA 51 VG, or PBS. Taking advantage of recombinant swine MHC class I molecules (SLAs), the peptide-SLA complex stability was measured for 198 IDO- or RhoC-derived 9-11mer peptides predicted to bind to SLA-1(*)04:01, -1(*)07:02, -2(*)04:01, -2(*)05:02, and/or -3(*)04:01. This identified 89 stable (t½ ≥ 0.5 h) peptide-SLA complexes. By IFN-γ release in PBMC cultures we monitored the vaccine-induced peptide-specific CTL responses, and found responses to both IDO- and RhoC-derived peptides across all groups with no adjuvant being superior. These findings support the further use of pigs as a large animal model for vaccine development against human cancer.

No MeSH data available.


Related in: MedlinePlus

Peptide-MHC class I binding stability as determined by SPA analysis. Stability of the predicted peptides in the 9-11mer peptide library with relevant SLA molecules measured by the Scintillation Proximity Assay. (A) SPA-determined half-life of peptides predicted as binders for each SLA molecule. (B) Individual peptides with SPA-determined half-life ≥0.5 h (stably binding) for each SLA molecule. Peptides derived from IDO and RhoC are shown in circles and diamonds, respectively.
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Figure 2: Peptide-MHC class I binding stability as determined by SPA analysis. Stability of the predicted peptides in the 9-11mer peptide library with relevant SLA molecules measured by the Scintillation Proximity Assay. (A) SPA-determined half-life of peptides predicted as binders for each SLA molecule. (B) Individual peptides with SPA-determined half-life ≥0.5 h (stably binding) for each SLA molecule. Peptides derived from IDO and RhoC are shown in circles and diamonds, respectively.

Mentions: In order to determine peptides capable of forming stable (t1∕2≥0.5 h) complexes with the relevant SLA amongst the 198 peptides, the half-life of the peptide-MHC class I binding was determined for each of the 244 predicted peptide-SLA complexes using SPA analysis. For the widely distributed SLA-1*04:01, a total of 12 IDO and RhoC-derived peptides formed stable complexes with this SLA molecule (Figure 2A), and especially IDO122 showed very high binding stability (t1∕2 = 30.2 h, Figure 2B). Also, 26 peptides formed stable complexes with the other well-distributed SLA-type amongst the pigs, namely SLA-1*07:02 (Figure 2A). Here, especially IDO21 (t1∕2 = 10.1 h) and IDO88 (t1∕2 = 24.0 h) were found to form highly stable complexes (Figure 2B). Strikingly, only five peptides were found to form stable complexes with SLA-3*04:01 in the SPA analysis (Figure 2A), and IDO105 was the only peptide being able to form a complex with a half-life longer than 1 h with this SLA molecule (t1∕2 = 14.7 h) (Figure 2B). A total of 24 peptides were shown to form stable complexes with SLA-2*05:02 (Figure 2A) with IDO16 (t1∕2 = 12.5 h) forming the most stable complex. IDO21 and IDO29 were both found to form stable complexes with SLA-2*04:01 exhibiting half-lives of 23.8 and 33.6 h, respectively (Figure 2B), and additional 20 peptides also formed stable complexes with SLA-2*04:01. To sum up, the SPA analysis revealed 89 stable peptide-SLA complexes (80 different peptides) from the 244 predicted high-affinity complexes (198 different peptides), and these were used in subsequent analyses of the CD8+ T-cell reactivity.


Establishing the pig as a large animal model for vaccine development against human cancer.

Overgaard NH, Frøsig TM, Welner S, Rasmussen M, Ilsøe M, Sørensen MR, Andersen MH, Buus S, Jungersen G - Front Genet (2015)

Peptide-MHC class I binding stability as determined by SPA analysis. Stability of the predicted peptides in the 9-11mer peptide library with relevant SLA molecules measured by the Scintillation Proximity Assay. (A) SPA-determined half-life of peptides predicted as binders for each SLA molecule. (B) Individual peptides with SPA-determined half-life ≥0.5 h (stably binding) for each SLA molecule. Peptides derived from IDO and RhoC are shown in circles and diamonds, respectively.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4584933&req=5

Figure 2: Peptide-MHC class I binding stability as determined by SPA analysis. Stability of the predicted peptides in the 9-11mer peptide library with relevant SLA molecules measured by the Scintillation Proximity Assay. (A) SPA-determined half-life of peptides predicted as binders for each SLA molecule. (B) Individual peptides with SPA-determined half-life ≥0.5 h (stably binding) for each SLA molecule. Peptides derived from IDO and RhoC are shown in circles and diamonds, respectively.
Mentions: In order to determine peptides capable of forming stable (t1∕2≥0.5 h) complexes with the relevant SLA amongst the 198 peptides, the half-life of the peptide-MHC class I binding was determined for each of the 244 predicted peptide-SLA complexes using SPA analysis. For the widely distributed SLA-1*04:01, a total of 12 IDO and RhoC-derived peptides formed stable complexes with this SLA molecule (Figure 2A), and especially IDO122 showed very high binding stability (t1∕2 = 30.2 h, Figure 2B). Also, 26 peptides formed stable complexes with the other well-distributed SLA-type amongst the pigs, namely SLA-1*07:02 (Figure 2A). Here, especially IDO21 (t1∕2 = 10.1 h) and IDO88 (t1∕2 = 24.0 h) were found to form highly stable complexes (Figure 2B). Strikingly, only five peptides were found to form stable complexes with SLA-3*04:01 in the SPA analysis (Figure 2A), and IDO105 was the only peptide being able to form a complex with a half-life longer than 1 h with this SLA molecule (t1∕2 = 14.7 h) (Figure 2B). A total of 24 peptides were shown to form stable complexes with SLA-2*05:02 (Figure 2A) with IDO16 (t1∕2 = 12.5 h) forming the most stable complex. IDO21 and IDO29 were both found to form stable complexes with SLA-2*04:01 exhibiting half-lives of 23.8 and 33.6 h, respectively (Figure 2B), and additional 20 peptides also formed stable complexes with SLA-2*04:01. To sum up, the SPA analysis revealed 89 stable peptide-SLA complexes (80 different peptides) from the 244 predicted high-affinity complexes (198 different peptides), and these were used in subsequent analyses of the CD8+ T-cell reactivity.

Bottom Line: Taking advantage of recombinant swine MHC class I molecules (SLAs), the peptide-SLA complex stability was measured for 198 IDO- or RhoC-derived 9-11mer peptides predicted to bind to SLA-1(*)04:01, -1(*)07:02, -2(*)04:01, -2(*)05:02, and/or -3(*)04:01.This identified 89 stable (t½ ≥ 0.5 h) peptide-SLA complexes.By IFN-γ release in PBMC cultures we monitored the vaccine-induced peptide-specific CTL responses, and found responses to both IDO- and RhoC-derived peptides across all groups with no adjuvant being superior.

View Article: PubMed Central - PubMed

Affiliation: Department of Immunology and Vaccinology, National Veterinary Institute, Technical University of Denmark Copenhagen, Denmark.

ABSTRACT
Immunotherapy has increased overall survival of metastatic cancer patients, and cancer antigens are promising vaccine targets. To fulfill the promise, appropriate tailoring of the vaccine formulations to mount in vivo cytotoxic T cell (CTL) responses toward co-delivered cancer antigens is essential. Previous development of therapeutic cancer vaccines has largely been based on studies in mice, and the majority of these candidate vaccines failed to induce therapeutic responses in the subsequent human clinical trials. Given that antigen dose and vaccine volume in pigs are translatable to humans and the porcine immunome is closer related to the human counterpart, we here introduce pigs as a supplementary large animal model for human cancer vaccine development. IDO and RhoC, both important in human cancer development and progression, were used as vaccine targets and 12 pigs were immunized with overlapping 20mer peptides spanning the entire porcine IDO and RhoC sequences formulated in CTL-inducing adjuvants: CAF09, CASAC, Montanide ISA 51 VG, or PBS. Taking advantage of recombinant swine MHC class I molecules (SLAs), the peptide-SLA complex stability was measured for 198 IDO- or RhoC-derived 9-11mer peptides predicted to bind to SLA-1(*)04:01, -1(*)07:02, -2(*)04:01, -2(*)05:02, and/or -3(*)04:01. This identified 89 stable (t½ ≥ 0.5 h) peptide-SLA complexes. By IFN-γ release in PBMC cultures we monitored the vaccine-induced peptide-specific CTL responses, and found responses to both IDO- and RhoC-derived peptides across all groups with no adjuvant being superior. These findings support the further use of pigs as a large animal model for vaccine development against human cancer.

No MeSH data available.


Related in: MedlinePlus