Identification of murine cytotoxic T-lymphocyte epitopes of bovine herpesvirus 1
Identifieur interne : 000474 ( Istex/Corpus ); précédent : 000473; suivant : 000475Identification of murine cytotoxic T-lymphocyte epitopes of bovine herpesvirus 1
Auteurs : Douglas S. Zatechka Jr. ; Nagendra R. Hegde ; Kandasamy Hariharan ; S. SrikumaranSource :
- Vaccine [ 0264-410X ] ; 1999.
English descriptors
- Teeft :
- Adjuvant, Allele, Aspms, Assay, Bovine, Bovine herpesvirus, Cell epitopes, Ctls, Cytotoxic, Cytotoxicity assay, Epitope, Error bars, Glycoprotein, Herpesvirus, Immunization, Immunol, Individual peptides, Lymphocyte, Lysed, Lysis, Maximum release, Molecule, Murine, Native peptides, Particular class, Peptide, Peptide motif, Peptide motifs, Recombinant, Spontaneous release, Standard deviations, Syngeneic, Synthetic peptides, Target cells, Vaccine, Viral, Viral immunol, Viral peptides, Virol, Zatechka.
Abstract
Abstract: Major histocompatibility complex (MHC) class I molecules present endogenously derived viral peptides to CD8+ cytotoxic T-lymphocytes (CTLs). The objective of this study was to identify the H-2Dd- and H-2Kd-restricted CTL epitopes of bovine herpesvirus 1 (BHV-1), based on the allele-specific peptide motifs (ASPMs) of the above class I molecules. Nine sequences conforming to the H-2Dd and H-2Kd ASPMs were identified on BHV-1 proteins, and the respective peptides were synthesized. Five of these peptides exhibited moderate to strong binding to the Dd molecule. CTLs generated by BALB/c mice immunized with BHV-1 proteins emulsified in a suitable adjuvant effectively lysed peptide-pulsed syngeneic targets, indicating that these epitopes were generated in vivo. Mice immunized with these peptides emulsified in a suitable adjuvant also developed anti-BHV-1 CTLs. These CTLs identified three veritable CTL epitopes among the “potential epitopes” synthesized based on the ASPMs. The elucidation of the CTL epitopes of BHV-1 should aid in the development of efficacious vaccines against this virus.
Url:
DOI: 10.1016/S0264-410X(98)00251-5
Links to Exploration step
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<front><div type="abstract" xml:lang="en">Abstract: Major histocompatibility complex (MHC) class I molecules present endogenously derived viral peptides to CD8+ cytotoxic T-lymphocytes (CTLs). The objective of this study was to identify the H-2Dd- and H-2Kd-restricted CTL epitopes of bovine herpesvirus 1 (BHV-1), based on the allele-specific peptide motifs (ASPMs) of the above class I molecules. Nine sequences conforming to the H-2Dd and H-2Kd ASPMs were identified on BHV-1 proteins, and the respective peptides were synthesized. Five of these peptides exhibited moderate to strong binding to the Dd molecule. CTLs generated by BALB/c mice immunized with BHV-1 proteins emulsified in a suitable adjuvant effectively lysed peptide-pulsed syngeneic targets, indicating that these epitopes were generated in vivo. Mice immunized with these peptides emulsified in a suitable adjuvant also developed anti-BHV-1 CTLs. These CTLs identified three veritable CTL epitopes among the “potential epitopes” synthesized based on the ASPMs. The elucidation of the CTL epitopes of BHV-1 should aid in the development of efficacious vaccines against this virus.</div>
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<note type="content">Fig. 1: The ability of the synthetic peptides to bind class I molecules. BALB/3T3 cells were treated with citrate–phosphate buffer, pH 3.3, for 20 s. Dissociation of the native peptides (stripping) was monitored by flow cytometry using antibodies that bind H-2d molecules only in the presence of bound peptides (TIB-126) or irrespective of peptide binding (HB87). The ability of the synthetic peptides to individually reconstitute the TIB-126 epitope was tested by incubating the stripped cells with peptides, in the presence of human β2-microglobulin. Results are expressed as fluorescence index (FI) calculated as follows: FI=(Mean sample fluorescence−Mean background fluorescence)/(Mean background fluorescence)×100, where background refers to peptide-stripped cells and sample refers to peptide-pulsed cells. The error bars indicate standard deviations of the means.</note>
<note type="content">Fig. 2: Immunization of mice with BHV-1 protein or peptides induces BHV-1-specific CTLs. Mice were immunized and boosted with protein (Panel A) or peptides (Panel B), in PROVAX™ or TiterMax, and lymphocytes were harvested from spleen (Panel A) or popliteal lymph nodes (Panel B), respectively. In vitro restimulated effectors were tested against syngeneic targets either pulsed with a cocktail of peptides or not pulsed with the peptides, at the indicated E:T ratios, in a standard 51Cr-release assay. The results are presented as % specific lysis, calculated as follows: % specific lysis=(Sample 51Cr release−Spontaneous 51Cr release)/(Maximum 51Cr release−Spontaneous 51Cr release) where spontaneous and maximum release refer to no effector and Triton X-100-treated samples, respectively. Spontaneous release was less than 20% of maximum release in all cases. The error bars indicate standard deviations of the means.</note>
<note type="content">Fig. 3: Identification of H-2Dd-restricted CTL epitopes of BHV-1. CTLs derived as described in Fig. 2B, were tested in a 51Cr-release assay using syngeneic cells pulsed with individual peptides at the indicated E:T ratios. The negative control peptide was a nonamer from BVDV gp53. The spontaneous release was less than 18% of maximum release in all cases. The error bars indicate standard deviations of the means. The means were statistically significant when no antigen was compared individually with peptides 1, 6 and 7, but not significant between no antigen and peptide (p<0.05).</note>
<note type="content">Fig. 4: BHV-1-specific CTLs are H-2Dd-restricted. CTLs obtained as described in Fig. 2, were tested in a 51Cr-release assay using targets pulsed with a pool of peptides 1, 6 and 7. The results are presented as described in Fig. 2. Spontaneous release was less than 12% of maximum release in all cases. (A) CTLs are class I-restricted. BALB/3T3 cells that do not express class II molecules were used as targets. (B) CTLs are H-2Dd-restricted. Syngeneic (P815, H-2d) and allogeneic (EL-4, H-2b) targets were used. The error bars indicate standard deviations of the means.</note>
<note type="content">Table 1: The allele-specific peptide motifs (ASPMs) of the MHC class I products utilized</note>
<note type="content">Table 2: Peptides synthesized in this study</note>
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<author xml:id="author-0001"><persName><forename type="first">Nagendra R.</forename>
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<note type="correspondence"><p>Corresponding author. Tel.: +1-402-472-3319; fax: +1-402-472-9690</p>
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<abstract xml:lang="en"><p>Abstract: Major histocompatibility complex (MHC) class I molecules present endogenously derived viral peptides to CD8+ cytotoxic T-lymphocytes (CTLs). The objective of this study was to identify the H-2Dd- and H-2Kd-restricted CTL epitopes of bovine herpesvirus 1 (BHV-1), based on the allele-specific peptide motifs (ASPMs) of the above class I molecules. Nine sequences conforming to the H-2Dd and H-2Kd ASPMs were identified on BHV-1 proteins, and the respective peptides were synthesized. Five of these peptides exhibited moderate to strong binding to the Dd molecule. CTLs generated by BALB/c mice immunized with BHV-1 proteins emulsified in a suitable adjuvant effectively lysed peptide-pulsed syngeneic targets, indicating that these epitopes were generated in vivo. Mice immunized with these peptides emulsified in a suitable adjuvant also developed anti-BHV-1 CTLs. These CTLs identified three veritable CTL epitopes among the “potential epitopes” synthesized based on the ASPMs. The elucidation of the CTL epitopes of BHV-1 should aid in the development of efficacious vaccines against this virus.</p>
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<head><ce:title>Identification of murine cytotoxic T-lymphocyte epitopes of bovine herpesvirus 1</ce:title>
<ce:author-group><ce:author><ce:given-name>Douglas S.</ce:given-name>
<ce:surname>Zatechka</ce:surname>
<ce:suffix>Jr.</ce:suffix>
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<ce:author><ce:given-name>Nagendra R.</ce:given-name>
<ce:surname>Hegde</ce:surname>
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<ce:author><ce:given-name>Kandasamy</ce:given-name>
<ce:surname>Hariharan</ce:surname>
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<ce:author><ce:given-name>S.</ce:given-name>
<ce:surname>Srikumaran</ce:surname>
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<ce:cross-ref refid="CORR1">*</ce:cross-ref>
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<ce:textfn>Department of Veterinary and Biomedical Sciences, University of Nebraska-Lincoln, Fair Street and East Campus Loop, Lincoln, NE 68583-0905, USA</ce:textfn>
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<ce:affiliation id="AFF2"><ce:label>b</ce:label>
<ce:textfn>IDEC Pharmaceuticals Corporation, San Diego, CA 92121, USA</ce:textfn>
</ce:affiliation>
<ce:correspondence id="CORR1"><ce:label>*</ce:label>
<ce:text>Corresponding author. Tel.: +1-402-472-3319; fax: +1-402-472-9690</ce:text>
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<ce:abstract><ce:section-title>Abstract</ce:section-title>
<ce:abstract-sec><ce:simple-para>Major histocompatibility complex (MHC) class I molecules present endogenously derived viral peptides to CD8<ce:sup>+</ce:sup>
cytotoxic T-lymphocytes (CTLs). The objective of this study was to identify the H-2D<ce:sup>d</ce:sup>
- and H-2K<ce:sup>d</ce:sup>
-restricted CTL epitopes of bovine herpesvirus 1 (BHV-1), based on the allele-specific peptide motifs (ASPMs) of the above class I molecules. Nine sequences conforming to the H-2D<ce:sup>d</ce:sup>
and H-2K<ce:sup>d</ce:sup>
ASPMs were identified on BHV-1 proteins, and the respective peptides were synthesized. Five of these peptides exhibited moderate to strong binding to the D<ce:sup>d</ce:sup>
molecule. CTLs generated by BALB/c mice immunized with BHV-1 proteins emulsified in a suitable adjuvant effectively lysed peptide-pulsed syngeneic targets, indicating that these epitopes were generated in vivo. Mice immunized with these peptides emulsified in a suitable adjuvant also developed anti-BHV-1 CTLs. These CTLs identified three veritable CTL epitopes among the “potential epitopes” synthesized based on the ASPMs. The elucidation of the CTL epitopes of BHV-1 should aid in the development of efficacious vaccines against this virus.</ce:simple-para>
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<ce:keywords class="keyword"><ce:section-title>Keywords</ce:section-title>
<ce:keyword><ce:text>Allele-specific peptide motif</ce:text>
</ce:keyword>
<ce:keyword><ce:text>BHV-1</ce:text>
</ce:keyword>
<ce:keyword><ce:text>CTL epitope</ce:text>
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<name type="personal"><namePart type="given">Douglas S.</namePart>
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<affiliation>Department of Veterinary and Biomedical Sciences, University of Nebraska-Lincoln, Fair Street and East Campus Loop, Lincoln, NE 68583-0905, USA</affiliation>
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<abstract lang="en">Abstract: Major histocompatibility complex (MHC) class I molecules present endogenously derived viral peptides to CD8+ cytotoxic T-lymphocytes (CTLs). The objective of this study was to identify the H-2Dd- and H-2Kd-restricted CTL epitopes of bovine herpesvirus 1 (BHV-1), based on the allele-specific peptide motifs (ASPMs) of the above class I molecules. Nine sequences conforming to the H-2Dd and H-2Kd ASPMs were identified on BHV-1 proteins, and the respective peptides were synthesized. Five of these peptides exhibited moderate to strong binding to the Dd molecule. CTLs generated by BALB/c mice immunized with BHV-1 proteins emulsified in a suitable adjuvant effectively lysed peptide-pulsed syngeneic targets, indicating that these epitopes were generated in vivo. Mice immunized with these peptides emulsified in a suitable adjuvant also developed anti-BHV-1 CTLs. These CTLs identified three veritable CTL epitopes among the “potential epitopes” synthesized based on the ASPMs. The elucidation of the CTL epitopes of BHV-1 should aid in the development of efficacious vaccines against this virus.</abstract>
<note type="content">Fig. 1: The ability of the synthetic peptides to bind class I molecules. BALB/3T3 cells were treated with citrate–phosphate buffer, pH 3.3, for 20 s. Dissociation of the native peptides (stripping) was monitored by flow cytometry using antibodies that bind H-2d molecules only in the presence of bound peptides (TIB-126) or irrespective of peptide binding (HB87). The ability of the synthetic peptides to individually reconstitute the TIB-126 epitope was tested by incubating the stripped cells with peptides, in the presence of human β2-microglobulin. Results are expressed as fluorescence index (FI) calculated as follows: FI=(Mean sample fluorescence−Mean background fluorescence)/(Mean background fluorescence)×100, where background refers to peptide-stripped cells and sample refers to peptide-pulsed cells. The error bars indicate standard deviations of the means.</note>
<note type="content">Fig. 2: Immunization of mice with BHV-1 protein or peptides induces BHV-1-specific CTLs. Mice were immunized and boosted with protein (Panel A) or peptides (Panel B), in PROVAX™ or TiterMax, and lymphocytes were harvested from spleen (Panel A) or popliteal lymph nodes (Panel B), respectively. In vitro restimulated effectors were tested against syngeneic targets either pulsed with a cocktail of peptides or not pulsed with the peptides, at the indicated E:T ratios, in a standard 51Cr-release assay. The results are presented as % specific lysis, calculated as follows: % specific lysis=(Sample 51Cr release−Spontaneous 51Cr release)/(Maximum 51Cr release−Spontaneous 51Cr release) where spontaneous and maximum release refer to no effector and Triton X-100-treated samples, respectively. Spontaneous release was less than 20% of maximum release in all cases. The error bars indicate standard deviations of the means.</note>
<note type="content">Fig. 3: Identification of H-2Dd-restricted CTL epitopes of BHV-1. CTLs derived as described in Fig. 2B, were tested in a 51Cr-release assay using syngeneic cells pulsed with individual peptides at the indicated E:T ratios. The negative control peptide was a nonamer from BVDV gp53. The spontaneous release was less than 18% of maximum release in all cases. The error bars indicate standard deviations of the means. The means were statistically significant when no antigen was compared individually with peptides 1, 6 and 7, but not significant between no antigen and peptide (p<0.05).</note>
<note type="content">Fig. 4: BHV-1-specific CTLs are H-2Dd-restricted. CTLs obtained as described in Fig. 2, were tested in a 51Cr-release assay using targets pulsed with a pool of peptides 1, 6 and 7. The results are presented as described in Fig. 2. Spontaneous release was less than 12% of maximum release in all cases. (A) CTLs are class I-restricted. BALB/3T3 cells that do not express class II molecules were used as targets. (B) CTLs are H-2Dd-restricted. Syngeneic (P815, H-2d) and allogeneic (EL-4, H-2b) targets were used. The error bars indicate standard deviations of the means.</note>
<note type="content">Table 1: The allele-specific peptide motifs (ASPMs) of the MHC class I products utilized</note>
<note type="content">Table 2: Peptides synthesized in this study</note>
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