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Comparison of influenza and SIV specific CD8 T cell responses in macaques.

Jegaskanda S, Reece JC, De Rose R, Stambas J, Sullivan L, Brooks AG, Kent SJ, Sexton A - PLoS ONE (2012)

Bottom Line: We recently developed an influenza-SIV vaccination model of pigtail macaques (Macaca nemestrina) and used this to study both influenza-specific and SIV-specific CD8(+) T-cells in 39 pigtail macaques expressing the common Mane-A*10(+) (Mane-A01*084) MHC-I allele.In contrast, within weeks following active SIV infection, SIV-specific CD8(+) effector T-cells expressed fewer cytokines/degranulation markers and had a lower avidity compared to influenza specific CD8(+) T-cells.This contrasted with the effector SIV-specific CD8(+) T-cells following SIV infection which expressed significantly higher amounts of PD-1 and lower amounts of CD28.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology and Immunology, University of Melbourne, Melbourne, Victoria, Australia.

ABSTRACT
Macaques are a potentially useful non-human primate model to compare memory T-cell immunity to acute virus pathogens such as influenza virus and effector T-cell responses to chronic viral pathogens such as SIV. However, immunological reagents to study influenza CD8(+) T-cell responses in the macaque model are limited. We recently developed an influenza-SIV vaccination model of pigtail macaques (Macaca nemestrina) and used this to study both influenza-specific and SIV-specific CD8(+) T-cells in 39 pigtail macaques expressing the common Mane-A*10(+) (Mane-A01*084) MHC-I allele. To perform comparative studies between influenza and SIV responses a common influenza nucleoprotein-specific CD8(+) T-cell response was mapped to a minimal epitope (termed RA9), MHC-restricted to Mane-A*10 and an MHC tetramer developed to study this response. Influenza-specific memory CD8(+) T-cell response maintained a highly functional profile in terms of multitude of effector molecule expression (CD107a, IFN-γ, TNF-α, MIP-1β and IL-2) and showed high avidity even in the setting of SIV infection. In contrast, within weeks following active SIV infection, SIV-specific CD8(+) effector T-cells expressed fewer cytokines/degranulation markers and had a lower avidity compared to influenza specific CD8(+) T-cells. Further, the influenza specific memory CD8 T-cell response retained stable expression of the exhaustion marker programmed death-marker-1 (PD-1) and co-stimulatory molecule CD28 following infection with SIV. This contrasted with the effector SIV-specific CD8(+) T-cells following SIV infection which expressed significantly higher amounts of PD-1 and lower amounts of CD28. Our results suggest that strategies to maintain a more functional CD8(+) T-cell response, profile may assist in controlling HIV disease.

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Comparison of tetramer positive influenza and SIV-specific CD8 T cell responses following recombinant influenza-SIV vaccination.Polyfunctional-tetramer ICS assay was performed on whole blood from day 0 post SIV infection stimulating with either RA9 or KVA10 peptides. (A) Summary of functional profile; the proportions of the number of effector molecules produced by antigen specific CD8 T cells when stimulated with either RA9 or KVA10 peptide. (B) The frequency of total effector molecules produced by RA9 (black) and KVA10 (white) specific CD8 T cells. Results are shown for 2 separate animals. (C) Comparison of the kinetics of effector molecule for RA9 (filled) versus KVA10 (open) at day 5 post SIV infection.
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pone-0032431-g006: Comparison of tetramer positive influenza and SIV-specific CD8 T cell responses following recombinant influenza-SIV vaccination.Polyfunctional-tetramer ICS assay was performed on whole blood from day 0 post SIV infection stimulating with either RA9 or KVA10 peptides. (A) Summary of functional profile; the proportions of the number of effector molecules produced by antigen specific CD8 T cells when stimulated with either RA9 or KVA10 peptide. (B) The frequency of total effector molecules produced by RA9 (black) and KVA10 (white) specific CD8 T cells. Results are shown for 2 separate animals. (C) Comparison of the kinetics of effector molecule for RA9 (filled) versus KVA10 (open) at day 5 post SIV infection.

Mentions: Having optimized the use of the Mane-A*10-RA9 tetramer in the ICS assay we used this assay to determine the polyfunctionality of influenza- and SIV-specific CD8 T cell responses after influenza-SIV vaccination. As seen with the standard ICS assay (Figure 2) the overall pattern of effector molecule expression from CD8 T cells in response to either influenza RA9 or SIV KVA10 were similar (Figure 6a and 6b). This is consistent with the epitopes being expressed from the same vector and consistent with other studies using vaccinia vectors [49], [50]. Interestingly, the incorporation of tetramer in the ICS assay also allowed us to determine that close to half of the RA9- and KVA10-specific CD8 T cells did not produce any of the studied effectors molecules (as shown by pale section of pie chart in Fig. 6a), with 3–15% producing one effector molecule and very few cells (<1%) producing all 4 effector molecules. These studies were repeated in another 3 animals with similar results, where the most common number of effector molecule expressed was none, 11–15% of cells expressed one and <1% producing all 4 effector molecules. The pattern of expression of effector molecule expression was also similar across another 3 animals, with MIP1β the least commonly expressed (<2% of total) and IFNγ, TNFα and CD107a all of reasonable expression levels (47.9%, 20.2% and 31.4% of total respectively).


Comparison of influenza and SIV specific CD8 T cell responses in macaques.

Jegaskanda S, Reece JC, De Rose R, Stambas J, Sullivan L, Brooks AG, Kent SJ, Sexton A - PLoS ONE (2012)

Comparison of tetramer positive influenza and SIV-specific CD8 T cell responses following recombinant influenza-SIV vaccination.Polyfunctional-tetramer ICS assay was performed on whole blood from day 0 post SIV infection stimulating with either RA9 or KVA10 peptides. (A) Summary of functional profile; the proportions of the number of effector molecules produced by antigen specific CD8 T cells when stimulated with either RA9 or KVA10 peptide. (B) The frequency of total effector molecules produced by RA9 (black) and KVA10 (white) specific CD8 T cells. Results are shown for 2 separate animals. (C) Comparison of the kinetics of effector molecule for RA9 (filled) versus KVA10 (open) at day 5 post SIV infection.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3293803&req=5

pone-0032431-g006: Comparison of tetramer positive influenza and SIV-specific CD8 T cell responses following recombinant influenza-SIV vaccination.Polyfunctional-tetramer ICS assay was performed on whole blood from day 0 post SIV infection stimulating with either RA9 or KVA10 peptides. (A) Summary of functional profile; the proportions of the number of effector molecules produced by antigen specific CD8 T cells when stimulated with either RA9 or KVA10 peptide. (B) The frequency of total effector molecules produced by RA9 (black) and KVA10 (white) specific CD8 T cells. Results are shown for 2 separate animals. (C) Comparison of the kinetics of effector molecule for RA9 (filled) versus KVA10 (open) at day 5 post SIV infection.
Mentions: Having optimized the use of the Mane-A*10-RA9 tetramer in the ICS assay we used this assay to determine the polyfunctionality of influenza- and SIV-specific CD8 T cell responses after influenza-SIV vaccination. As seen with the standard ICS assay (Figure 2) the overall pattern of effector molecule expression from CD8 T cells in response to either influenza RA9 or SIV KVA10 were similar (Figure 6a and 6b). This is consistent with the epitopes being expressed from the same vector and consistent with other studies using vaccinia vectors [49], [50]. Interestingly, the incorporation of tetramer in the ICS assay also allowed us to determine that close to half of the RA9- and KVA10-specific CD8 T cells did not produce any of the studied effectors molecules (as shown by pale section of pie chart in Fig. 6a), with 3–15% producing one effector molecule and very few cells (<1%) producing all 4 effector molecules. These studies were repeated in another 3 animals with similar results, where the most common number of effector molecule expressed was none, 11–15% of cells expressed one and <1% producing all 4 effector molecules. The pattern of expression of effector molecule expression was also similar across another 3 animals, with MIP1β the least commonly expressed (<2% of total) and IFNγ, TNFα and CD107a all of reasonable expression levels (47.9%, 20.2% and 31.4% of total respectively).

Bottom Line: We recently developed an influenza-SIV vaccination model of pigtail macaques (Macaca nemestrina) and used this to study both influenza-specific and SIV-specific CD8(+) T-cells in 39 pigtail macaques expressing the common Mane-A*10(+) (Mane-A01*084) MHC-I allele.In contrast, within weeks following active SIV infection, SIV-specific CD8(+) effector T-cells expressed fewer cytokines/degranulation markers and had a lower avidity compared to influenza specific CD8(+) T-cells.This contrasted with the effector SIV-specific CD8(+) T-cells following SIV infection which expressed significantly higher amounts of PD-1 and lower amounts of CD28.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology and Immunology, University of Melbourne, Melbourne, Victoria, Australia.

ABSTRACT
Macaques are a potentially useful non-human primate model to compare memory T-cell immunity to acute virus pathogens such as influenza virus and effector T-cell responses to chronic viral pathogens such as SIV. However, immunological reagents to study influenza CD8(+) T-cell responses in the macaque model are limited. We recently developed an influenza-SIV vaccination model of pigtail macaques (Macaca nemestrina) and used this to study both influenza-specific and SIV-specific CD8(+) T-cells in 39 pigtail macaques expressing the common Mane-A*10(+) (Mane-A01*084) MHC-I allele. To perform comparative studies between influenza and SIV responses a common influenza nucleoprotein-specific CD8(+) T-cell response was mapped to a minimal epitope (termed RA9), MHC-restricted to Mane-A*10 and an MHC tetramer developed to study this response. Influenza-specific memory CD8(+) T-cell response maintained a highly functional profile in terms of multitude of effector molecule expression (CD107a, IFN-γ, TNF-α, MIP-1β and IL-2) and showed high avidity even in the setting of SIV infection. In contrast, within weeks following active SIV infection, SIV-specific CD8(+) effector T-cells expressed fewer cytokines/degranulation markers and had a lower avidity compared to influenza specific CD8(+) T-cells. Further, the influenza specific memory CD8 T-cell response retained stable expression of the exhaustion marker programmed death-marker-1 (PD-1) and co-stimulatory molecule CD28 following infection with SIV. This contrasted with the effector SIV-specific CD8(+) T-cells following SIV infection which expressed significantly higher amounts of PD-1 and lower amounts of CD28. Our results suggest that strategies to maintain a more functional CD8(+) T-cell response, profile may assist in controlling HIV disease.

Show MeSH
Related in: MedlinePlus