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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

Overview of the immunization strategy. Prior to initiation of the vaccine trial, candidate pigs were blood sampled and SLA-typed in order to select animals holding one or more of the following SLA alleles: SLA-1*04:01, SLA-1*07:02, SLA-2*04:01, SLA-2*05:02, and SLA-3*04:01. Pigs were then purchased and blood sampled at day −2 to determine the background level of IFN-γ. At day 0, pigs were primed with 58 20mer overlapping peptides in combination with either CAF09, CASAC, ISA 51VG or PBS. Blood samples were obtained at day 12, and all pigs were then boosted with another round of immunization at day 19. Blood samples were obtained three times following boost namely at day 33, 40, and 54.
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Figure 1: Overview of the immunization strategy. Prior to initiation of the vaccine trial, candidate pigs were blood sampled and SLA-typed in order to select animals holding one or more of the following SLA alleles: SLA-1*04:01, SLA-1*07:02, SLA-2*04:01, SLA-2*05:02, and SLA-3*04:01. Pigs were then purchased and blood sampled at day −2 to determine the background level of IFN-γ. At day 0, pigs were primed with 58 20mer overlapping peptides in combination with either CAF09, CASAC, ISA 51VG or PBS. Blood samples were obtained at day 12, and all pigs were then boosted with another round of immunization at day 19. Blood samples were obtained three times following boost namely at day 33, 40, and 54.

Mentions: Previous studies have confirmed the involvement of CD8+ T cells in anti-cancer immune reactivity (Klebanoff et al., 2005; Sørensen et al., 2011b; Andersen, 2012; Joyce and Fearon, 2015; Rosenberg and Restifo, 2015) and anti-cancer vaccines are generally administered with the aim of enhancing this antigen- specific T-cell reactivity. To establish the pig as a large animal model for human cancer vaccine development, we constructed a monitoring platform for vaccine-induced T-cell reactivity. First, candidate pigs were blood sampled and SLA-typed in order to choose animals holding the relevant SLA class I molecules (Figure 1). Second, identification of proteins relevant for vaccination and prediction of candidate CD8+ T cell epitopes from the full protein sequence were carried out. The in vitro stability of the candidate T cell epitopes in complex with relevant SLA class I molecule was then examined. Pigs were immunized with 20mer overlapping peptides and blood sampled at various time points pre- and post-immunization in order to monitor the T-cell reactivity ex vivo (Figure 1). To increase the knowledge obtained from this study, we stratified the pigs in four groups based on their SLA-profile and immunized each group with peptides in combination with an adjuvant system or PBS.


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)

Overview of the immunization strategy. Prior to initiation of the vaccine trial, candidate pigs were blood sampled and SLA-typed in order to select animals holding one or more of the following SLA alleles: SLA-1*04:01, SLA-1*07:02, SLA-2*04:01, SLA-2*05:02, and SLA-3*04:01. Pigs were then purchased and blood sampled at day −2 to determine the background level of IFN-γ. At day 0, pigs were primed with 58 20mer overlapping peptides in combination with either CAF09, CASAC, ISA 51VG or PBS. Blood samples were obtained at day 12, and all pigs were then boosted with another round of immunization at day 19. Blood samples were obtained three times following boost namely at day 33, 40, and 54.
© Copyright Policy
Related In: Results  -  Collection

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

Figure 1: Overview of the immunization strategy. Prior to initiation of the vaccine trial, candidate pigs were blood sampled and SLA-typed in order to select animals holding one or more of the following SLA alleles: SLA-1*04:01, SLA-1*07:02, SLA-2*04:01, SLA-2*05:02, and SLA-3*04:01. Pigs were then purchased and blood sampled at day −2 to determine the background level of IFN-γ. At day 0, pigs were primed with 58 20mer overlapping peptides in combination with either CAF09, CASAC, ISA 51VG or PBS. Blood samples were obtained at day 12, and all pigs were then boosted with another round of immunization at day 19. Blood samples were obtained three times following boost namely at day 33, 40, and 54.
Mentions: Previous studies have confirmed the involvement of CD8+ T cells in anti-cancer immune reactivity (Klebanoff et al., 2005; Sørensen et al., 2011b; Andersen, 2012; Joyce and Fearon, 2015; Rosenberg and Restifo, 2015) and anti-cancer vaccines are generally administered with the aim of enhancing this antigen- specific T-cell reactivity. To establish the pig as a large animal model for human cancer vaccine development, we constructed a monitoring platform for vaccine-induced T-cell reactivity. First, candidate pigs were blood sampled and SLA-typed in order to choose animals holding the relevant SLA class I molecules (Figure 1). Second, identification of proteins relevant for vaccination and prediction of candidate CD8+ T cell epitopes from the full protein sequence were carried out. The in vitro stability of the candidate T cell epitopes in complex with relevant SLA class I molecule was then examined. Pigs were immunized with 20mer overlapping peptides and blood sampled at various time points pre- and post-immunization in order to monitor the T-cell reactivity ex vivo (Figure 1). To increase the knowledge obtained from this study, we stratified the pigs in four groups based on their SLA-profile and immunized each group with peptides in combination with an adjuvant system or PBS.

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