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Tattoo Delivery of a Semliki Forest Virus-Based Vaccine Encoding Human Papillomavirus E6 and E7.

van de Wall S, Walczak M, van Rooij N, Hoogeboom BN, Meijerhof T, Nijman HW, Daemen T - Vaccines (Basel) (2015)

Bottom Line: Tattoo injection of rSFV particles resulted in antigen expression in both the skin and draining lymph nodes.Strikingly, despite the observed lower overall transgene expression, SFVeE6,7 delivered via tattoo injection resulted in higher or equal levels of immune responses as compared to intramuscular injection.The intrinsic immunogenic potential of tattooing provides a benefit for immunotherapy based on an alphavirus.

View Article: PubMed Central - PubMed

Affiliation: Department of Medical Microbiology, University of Groningen, University Medical Center Groningen, HPC EB88, PO Box 30.001, 9700 RB Groningen, The Netherlands. m.n.s.van.de.wall@umcg.nl.

ABSTRACT
The skin is an attractive organ for immunization because of the presence of antigen-presenting cells. Intradermal delivery via tattooing has demonstrated superior vaccine immunogenicity of DNA vaccines in comparison to conventional delivery methods. In this study, we explored the efficacy of tattoo injection of a tumor vaccine based on recombinant Semliki Forest virus replicon particles (rSFV) targeting human papillomavirus (HPV). Tattoo injection of rSFV particles resulted in antigen expression in both the skin and draining lymph nodes. In comparison with intramuscular injection, the overall antigen expression determined at the site of administration and draining lymph nodes was 10-fold lower upon tattoo injection. Delivery of SFV particles encoding the E6 and E7 antigens of human papillomavirus type 16 (SFVeE6,7) via tattooing resulted in HPV-specific cytotoxic T cells and in vivo therapeutic antitumor response. Strikingly, despite the observed lower overall transgene expression, SFVeE6,7 delivered via tattoo injection resulted in higher or equal levels of immune responses as compared to intramuscular injection. The intrinsic immunogenic potential of tattooing provides a benefit for immunotherapy based on an alphavirus.

No MeSH data available.


Related in: MedlinePlus

SFVeE6,7 tattoo injections induce strong T cell responses. Mice were tattooed or intramuscularly immunized with 5 × 106 i.u. of SFVeE6,7 and boosted 14 days later using the same or alternative delivery route. Ten days after the booster immunization animals were sacrificed. Freshly isolated (a) and 7-day in vitro restimulated spleen cells (b) were stained with MHC class I tetramers and anti-CD8 antibodies and analyzed by flow cytometry. The percentages of tetramer-positive cells within the CD8 T cells are shown. Activity of restimulated spleen and inguinal lymph node cells were analyzed in regular (c); only spleen cells) and micro CTL assay (d; both spleen and lymph node cells). The frequencies of IFNγ-producing cells, both in the spleens and inguinal lymph nodes, were determined using Elispot assay. Results are expressed as number of IFNγ-producing cells per 106 splenocytes (e) or per 106 lymph node cells (f). Each dot represents an individual mouse; a–d—data from a representative experiment out of four (three mice/group); e—pooled data from four experiments; f—data from one experiment. E:T ratio—effector:target ratio. * p < 0.05 as compared with the im group.
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vaccines-03-00221-f002: SFVeE6,7 tattoo injections induce strong T cell responses. Mice were tattooed or intramuscularly immunized with 5 × 106 i.u. of SFVeE6,7 and boosted 14 days later using the same or alternative delivery route. Ten days after the booster immunization animals were sacrificed. Freshly isolated (a) and 7-day in vitro restimulated spleen cells (b) were stained with MHC class I tetramers and anti-CD8 antibodies and analyzed by flow cytometry. The percentages of tetramer-positive cells within the CD8 T cells are shown. Activity of restimulated spleen and inguinal lymph node cells were analyzed in regular (c); only spleen cells) and micro CTL assay (d; both spleen and lymph node cells). The frequencies of IFNγ-producing cells, both in the spleens and inguinal lymph nodes, were determined using Elispot assay. Results are expressed as number of IFNγ-producing cells per 106 splenocytes (e) or per 106 lymph node cells (f). Each dot represents an individual mouse; a–d—data from a representative experiment out of four (three mice/group); e—pooled data from four experiments; f—data from one experiment. E:T ratio—effector:target ratio. * p < 0.05 as compared with the im group.

Mentions: Mice were immunized by tattooing or intramuscular injection with 5 × 106 i.u. of SFVeE6,7 and boosted 14 days later using the same or the alternative route of administration. Animals were sacrificed ten days after the booster immunization. The frequency of HPV-specific CTLs was determined by direct staining of freshly isolated spleen cells with MHC class I tetramers and anti-CD8 antibodies. SFVeE6,7 tattoo prime and boost resulted in induction of HPV-specific CD8 T cells (approx. 0.8% RAHYNIVTF-specific cells in CD8 T cells; Figure 2a). Other immunization protocols resulted in comparable frequencies of antigen specific CD8 T cells. Tetramer staining after 7-day in vitro restimulation showed that CTLs induced in vivo with SFVeE6,7 tattooing readily proliferated in vitro, resulted in 50%–80% RAHYNIVTF-specific CD8 T cells (Figure 2b). These results were reflected in high CTL lytic activity of these cells as measured in a bulk 51Cr-release assay (Figure 2c). All immunization protocols used resulted in similar high levels of specific cytolysis (approx. 75% at an effector:target ratio of 30:1).


Tattoo Delivery of a Semliki Forest Virus-Based Vaccine Encoding Human Papillomavirus E6 and E7.

van de Wall S, Walczak M, van Rooij N, Hoogeboom BN, Meijerhof T, Nijman HW, Daemen T - Vaccines (Basel) (2015)

SFVeE6,7 tattoo injections induce strong T cell responses. Mice were tattooed or intramuscularly immunized with 5 × 106 i.u. of SFVeE6,7 and boosted 14 days later using the same or alternative delivery route. Ten days after the booster immunization animals were sacrificed. Freshly isolated (a) and 7-day in vitro restimulated spleen cells (b) were stained with MHC class I tetramers and anti-CD8 antibodies and analyzed by flow cytometry. The percentages of tetramer-positive cells within the CD8 T cells are shown. Activity of restimulated spleen and inguinal lymph node cells were analyzed in regular (c); only spleen cells) and micro CTL assay (d; both spleen and lymph node cells). The frequencies of IFNγ-producing cells, both in the spleens and inguinal lymph nodes, were determined using Elispot assay. Results are expressed as number of IFNγ-producing cells per 106 splenocytes (e) or per 106 lymph node cells (f). Each dot represents an individual mouse; a–d—data from a representative experiment out of four (three mice/group); e—pooled data from four experiments; f—data from one experiment. E:T ratio—effector:target ratio. * p < 0.05 as compared with the im group.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4494346&req=5

vaccines-03-00221-f002: SFVeE6,7 tattoo injections induce strong T cell responses. Mice were tattooed or intramuscularly immunized with 5 × 106 i.u. of SFVeE6,7 and boosted 14 days later using the same or alternative delivery route. Ten days after the booster immunization animals were sacrificed. Freshly isolated (a) and 7-day in vitro restimulated spleen cells (b) were stained with MHC class I tetramers and anti-CD8 antibodies and analyzed by flow cytometry. The percentages of tetramer-positive cells within the CD8 T cells are shown. Activity of restimulated spleen and inguinal lymph node cells were analyzed in regular (c); only spleen cells) and micro CTL assay (d; both spleen and lymph node cells). The frequencies of IFNγ-producing cells, both in the spleens and inguinal lymph nodes, were determined using Elispot assay. Results are expressed as number of IFNγ-producing cells per 106 splenocytes (e) or per 106 lymph node cells (f). Each dot represents an individual mouse; a–d—data from a representative experiment out of four (three mice/group); e—pooled data from four experiments; f—data from one experiment. E:T ratio—effector:target ratio. * p < 0.05 as compared with the im group.
Mentions: Mice were immunized by tattooing or intramuscular injection with 5 × 106 i.u. of SFVeE6,7 and boosted 14 days later using the same or the alternative route of administration. Animals were sacrificed ten days after the booster immunization. The frequency of HPV-specific CTLs was determined by direct staining of freshly isolated spleen cells with MHC class I tetramers and anti-CD8 antibodies. SFVeE6,7 tattoo prime and boost resulted in induction of HPV-specific CD8 T cells (approx. 0.8% RAHYNIVTF-specific cells in CD8 T cells; Figure 2a). Other immunization protocols resulted in comparable frequencies of antigen specific CD8 T cells. Tetramer staining after 7-day in vitro restimulation showed that CTLs induced in vivo with SFVeE6,7 tattooing readily proliferated in vitro, resulted in 50%–80% RAHYNIVTF-specific CD8 T cells (Figure 2b). These results were reflected in high CTL lytic activity of these cells as measured in a bulk 51Cr-release assay (Figure 2c). All immunization protocols used resulted in similar high levels of specific cytolysis (approx. 75% at an effector:target ratio of 30:1).

Bottom Line: Tattoo injection of rSFV particles resulted in antigen expression in both the skin and draining lymph nodes.Strikingly, despite the observed lower overall transgene expression, SFVeE6,7 delivered via tattoo injection resulted in higher or equal levels of immune responses as compared to intramuscular injection.The intrinsic immunogenic potential of tattooing provides a benefit for immunotherapy based on an alphavirus.

View Article: PubMed Central - PubMed

Affiliation: Department of Medical Microbiology, University of Groningen, University Medical Center Groningen, HPC EB88, PO Box 30.001, 9700 RB Groningen, The Netherlands. m.n.s.van.de.wall@umcg.nl.

ABSTRACT
The skin is an attractive organ for immunization because of the presence of antigen-presenting cells. Intradermal delivery via tattooing has demonstrated superior vaccine immunogenicity of DNA vaccines in comparison to conventional delivery methods. In this study, we explored the efficacy of tattoo injection of a tumor vaccine based on recombinant Semliki Forest virus replicon particles (rSFV) targeting human papillomavirus (HPV). Tattoo injection of rSFV particles resulted in antigen expression in both the skin and draining lymph nodes. In comparison with intramuscular injection, the overall antigen expression determined at the site of administration and draining lymph nodes was 10-fold lower upon tattoo injection. Delivery of SFV particles encoding the E6 and E7 antigens of human papillomavirus type 16 (SFVeE6,7) via tattooing resulted in HPV-specific cytotoxic T cells and in vivo therapeutic antitumor response. Strikingly, despite the observed lower overall transgene expression, SFVeE6,7 delivered via tattoo injection resulted in higher or equal levels of immune responses as compared to intramuscular injection. The intrinsic immunogenic potential of tattooing provides a benefit for immunotherapy based on an alphavirus.

No MeSH data available.


Related in: MedlinePlus