Quantification of the number of cytotoxic T cells specific for an immunodominant HCV-specific CTL epitope primed by DNA immunization
Identifieur interne : 001768 ( Istex/Corpus ); précédent : 001767; suivant : 001769Quantification of the number of cytotoxic T cells specific for an immunodominant HCV-specific CTL epitope primed by DNA immunization
Auteurs : Alexander Y. Lee ; Noelle K. Polakos ; Gillis R. Otten ; Jeffrey B. Ulmer ; Michael Houghton ; Xavier PaliardSource :
- Vaccine [ 0264-410X ] ; 2000.
English descriptors
- Teeft :
- Acute hepatitis, Assay, Cell epitopes, Cell repertoire, Cytometry, Cytotoxic, Cytotoxic activity, Days post booster, Elsevier science, Epitope, Gene expression, Hepatitis, Immune, Immune response, Immune responses, Immunization, Important role, Individual mice, Individual peptides, Intracellular, Intracellular staining, Mouse, Natural history, Ns5a, Ns5b, Peptide, Peptide pool, Plasmid, Release assay, Restimulated, Restimulation, Spleen, Spleen cells, Svbalb target cells, Target cells, Unpublished observations, Vaccine, Vaccine strategies, Viral, Viral infection, Virol shirai, Virus infection.
Abstract
Abstract: Priming of strong cellular immune responses to hepatitis C (HCV) is thought to be important for eradication of infection. Although productive infection of HCV occurs only reproducibly in humans and chimpanzees, definition of HCV-specific T cell epitopes in mice is necessary to screen efficiently HCV vaccine strategies for their ability to prime cellular immune responses. Out of seven strains of mice screened for immunodominant CTL epitopes against HCV-1a Core, E2, NS5a and NS5b, only one epitope (p214K9) in only one mouse strain was identified. Enumeration of p214K9-specific CD8+ cells by flow cytometry revealed that the number of epitope specific CTL primed by ‘naked’ DNA immunization was lower than that reported during viral infection. The p214K9 epitope described here, combined with analysis of CTL responses by flow cytometry, should be instrumental in ranking various HCV vaccine strategies for their ability to prime CTL responses.
Url:
DOI: 10.1016/S0264-410X(99)00486-7
Links to Exploration step
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<front><div type="abstract" xml:lang="en">Abstract: Priming of strong cellular immune responses to hepatitis C (HCV) is thought to be important for eradication of infection. Although productive infection of HCV occurs only reproducibly in humans and chimpanzees, definition of HCV-specific T cell epitopes in mice is necessary to screen efficiently HCV vaccine strategies for their ability to prime cellular immune responses. Out of seven strains of mice screened for immunodominant CTL epitopes against HCV-1a Core, E2, NS5a and NS5b, only one epitope (p214K9) in only one mouse strain was identified. Enumeration of p214K9-specific CD8+ cells by flow cytometry revealed that the number of epitope specific CTL primed by ‘naked’ DNA immunization was lower than that reported during viral infection. The p214K9 epitope described here, combined with analysis of CTL responses by flow cytometry, should be instrumental in ranking various HCV vaccine strategies for their ability to prime CTL responses.</div>
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<note type="content">Fig. 1: Mapping of HCV-NS5a regions recognized by CD8+ T cells. Spleen cells from DNA immunized, VV boosted C3H/HeJ mice were cultured in media alone or restimulated ex vivo with peptide pools (4–6 peptides per pool) or individual peptides and stained for surface CD8 and intracellular IFN-γ with anti-CD8-FITC and anti-IFN-γ-PE, respectively. Lymphocytes were first gated by side vs forward scatter light and then for CD8-FITC. Plots show log fluorescence intensity for CD8 (FITC) and IFN-γ (PE) observed for a representative individual mouse.</note>
<note type="content">Fig. 2: P214K9-specific CD8+ T cells are cytotoxic. Spleen cells from DNA-NS5a immunized C3H/HeJ (H-2k) mice were pooled and restimulated in vitro with p214. Cultures were then tested for cytotoxic activity in a standard 51Cr release assay against L929 (H-2k) target cells sensitized with (■) p214, (▾) p214 K, (⋅) p214K9, (×) p214J, or, SvBalb (H-2d) target cells sensitized with (□) p214 or (○) p214K9.</note>
<note type="content">Table 1: Summary of HCV-specific CTL epitope mapping in various mouse strains by flow cytometry</note>
<note type="content">Table 2: p214K9-specific CTL in C3H/HeJ mice primed with DNA-NS5a and boosted with either DNA-NS5a or VV-NS5a</note>
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<note type="correspondence"><p>Corresponding author. Tel.: +1-510-923-3545; fax: +1-510-923-2586</p>
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<abstract xml:lang="en"><p>Abstract: Priming of strong cellular immune responses to hepatitis C (HCV) is thought to be important for eradication of infection. Although productive infection of HCV occurs only reproducibly in humans and chimpanzees, definition of HCV-specific T cell epitopes in mice is necessary to screen efficiently HCV vaccine strategies for their ability to prime cellular immune responses. Out of seven strains of mice screened for immunodominant CTL epitopes against HCV-1a Core, E2, NS5a and NS5b, only one epitope (p214K9) in only one mouse strain was identified. Enumeration of p214K9-specific CD8+ cells by flow cytometry revealed that the number of epitope specific CTL primed by ‘naked’ DNA immunization was lower than that reported during viral infection. The p214K9 epitope described here, combined with analysis of CTL responses by flow cytometry, should be instrumental in ranking various HCV vaccine strategies for their ability to prime CTL responses.</p>
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<head><ce:title>Quantification of the number of cytotoxic T cells specific for an immunodominant HCV-specific CTL epitope primed by DNA immunization</ce:title>
<ce:author-group><ce:author><ce:given-name>Alexander Y.</ce:given-name>
<ce:surname>Lee</ce:surname>
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<ce:author><ce:given-name>Noelle K.</ce:given-name>
<ce:surname>Polakos</ce:surname>
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<ce:author><ce:given-name>Gillis R.</ce:given-name>
<ce:surname>Otten</ce:surname>
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<ce:author><ce:given-name>Jeffrey B.</ce:given-name>
<ce:surname>Ulmer</ce:surname>
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<ce:author><ce:given-name>Michael</ce:given-name>
<ce:surname>Houghton</ce:surname>
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<ce:author><ce:given-name>Xavier</ce:given-name>
<ce:surname>Paliard</ce:surname>
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<ce:e-address>xavier_paliard@cc.chiron.com</ce:e-address>
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<ce:affiliation><ce:textfn>Chiron Corporation, 4560 Horton Street, Emeryville, CA 94608, USA</ce:textfn>
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<ce:text>Corresponding author. Tel.: +1-510-923-3545; fax: +1-510-923-2586</ce:text>
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<ce:abstract-sec><ce:simple-para>Priming of strong cellular immune responses to hepatitis C (HCV) is thought to be important for eradication of infection. Although productive infection of HCV occurs only reproducibly in humans and chimpanzees, definition of HCV-specific T cell epitopes in mice is necessary to screen efficiently HCV vaccine strategies for their ability to prime cellular immune responses. Out of seven strains of mice screened for immunodominant CTL epitopes against HCV-1a Core, E2, NS5a and NS5b, only one epitope (p214K9) in only one mouse strain was identified. Enumeration of p214K9-specific CD8+ cells by flow cytometry revealed that the number of epitope specific CTL primed by ‘naked’ DNA immunization was lower than that reported during viral infection. The p214K9 epitope described here, combined with analysis of CTL responses by flow cytometry, should be instrumental in ranking various HCV vaccine strategies for their ability to prime CTL responses.</ce:simple-para>
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<ce:keyword><ce:text>DNA immunization</ce:text>
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<abstract lang="en">Abstract: Priming of strong cellular immune responses to hepatitis C (HCV) is thought to be important for eradication of infection. Although productive infection of HCV occurs only reproducibly in humans and chimpanzees, definition of HCV-specific T cell epitopes in mice is necessary to screen efficiently HCV vaccine strategies for their ability to prime cellular immune responses. Out of seven strains of mice screened for immunodominant CTL epitopes against HCV-1a Core, E2, NS5a and NS5b, only one epitope (p214K9) in only one mouse strain was identified. Enumeration of p214K9-specific CD8+ cells by flow cytometry revealed that the number of epitope specific CTL primed by ‘naked’ DNA immunization was lower than that reported during viral infection. The p214K9 epitope described here, combined with analysis of CTL responses by flow cytometry, should be instrumental in ranking various HCV vaccine strategies for their ability to prime CTL responses.</abstract>
<note type="content">Fig. 1: Mapping of HCV-NS5a regions recognized by CD8+ T cells. Spleen cells from DNA immunized, VV boosted C3H/HeJ mice were cultured in media alone or restimulated ex vivo with peptide pools (4–6 peptides per pool) or individual peptides and stained for surface CD8 and intracellular IFN-γ with anti-CD8-FITC and anti-IFN-γ-PE, respectively. Lymphocytes were first gated by side vs forward scatter light and then for CD8-FITC. Plots show log fluorescence intensity for CD8 (FITC) and IFN-γ (PE) observed for a representative individual mouse.</note>
<note type="content">Fig. 2: P214K9-specific CD8+ T cells are cytotoxic. Spleen cells from DNA-NS5a immunized C3H/HeJ (H-2k) mice were pooled and restimulated in vitro with p214. Cultures were then tested for cytotoxic activity in a standard 51Cr release assay against L929 (H-2k) target cells sensitized with (■) p214, (▾) p214 K, (⋅) p214K9, (×) p214J, or, SvBalb (H-2d) target cells sensitized with (□) p214 or (○) p214K9.</note>
<note type="content">Table 1: Summary of HCV-specific CTL epitope mapping in various mouse strains by flow cytometry</note>
<note type="content">Table 2: p214K9-specific CTL in C3H/HeJ mice primed with DNA-NS5a and boosted with either DNA-NS5a or VV-NS5a</note>
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