Identification of HLA-A33-restricted CMV pp65 epitopes as common targets for CD8+ CMV-specific cytotoxic T lymphocytes

Jong Baeck Lim, Maurizio Provenzano, Oh Hun Kwon, Maria Bettinotti, Lorraine Caruccio, Dirk Nagorsen, David Stroncek

Research output: Contribution to journalArticle

Abstract

Objective. To identify an immunodominant HLA-A33-restricted epitope within the CMV matrix phosphoprotein 65 (pp65) that could be used to elicit CMV-specific CTLs. Methods. A computer algorithm was used to identify pp65 peptides that were expected to bind to HLA-A33. The peptides were screened for their ability to reactivate memory T lymphocytes from CMV-seropositive HLA-A33 donors by using quantitative real-time RT-PCR. The most promising peptides were then tested for their ability to expand a CD8+ population of HLA-A33 CTLs that produced interferon-γ (IFN-γ) and were cytotoxic to either peptide-loaded or CMV-infected target cells. Results. Sixteen out of 250 peptides were selected using a computer algorithm and peptide stimulation by 8 out of the 16 induced a significant quantity of IFN-γ mRNA transcript from CMV-seropositive HLA-A33 peripheral blood mononuclear cells. All the eight peptides induced consistent T-cell reactivation. One specifically, the peptide pp6591-100 (SVNVHNPTGR), proved to be more active. T cells in vitro sensitized for 2 or 3 weeks with pp6591-100 were cytotoxic to both HLA-A33 peptide-loaded and CMV-infected target cells. Conclusions. CMV pp65 91-100 is a new HLA-A33-restricted peptide that broadens the list of antigenic determinants that can be used for CMV adoptive immunotherapy.

Original languageEnglish (US)
Pages (from-to)296-307
Number of pages12
JournalExperimental Hematology
Volume34
Issue number3
DOIs
StatePublished - Mar 2006
Externally publishedYes

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HLA-A*33 antigen
Phosphoproteins
Cytotoxic T-Lymphocytes
Epitopes
Peptides
T-Lymphocytes
Interferons
Adoptive Immunotherapy

ASJC Scopus subject areas

  • Cancer Research
  • Cell Biology
  • Genetics
  • Hematology
  • Oncology
  • Transplantation

Cite this

Identification of HLA-A33-restricted CMV pp65 epitopes as common targets for CD8+ CMV-specific cytotoxic T lymphocytes. / Lim, Jong Baeck; Provenzano, Maurizio; Kwon, Oh Hun; Bettinotti, Maria; Caruccio, Lorraine; Nagorsen, Dirk; Stroncek, David.

In: Experimental Hematology, Vol. 34, No. 3, 03.2006, p. 296-307.

Research output: Contribution to journalArticle

Lim, Jong Baeck ; Provenzano, Maurizio ; Kwon, Oh Hun ; Bettinotti, Maria ; Caruccio, Lorraine ; Nagorsen, Dirk ; Stroncek, David. / Identification of HLA-A33-restricted CMV pp65 epitopes as common targets for CD8+ CMV-specific cytotoxic T lymphocytes. In: Experimental Hematology. 2006 ; Vol. 34, No. 3. pp. 296-307.
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abstract = "Objective. To identify an immunodominant HLA-A33-restricted epitope within the CMV matrix phosphoprotein 65 (pp65) that could be used to elicit CMV-specific CTLs. Methods. A computer algorithm was used to identify pp65 peptides that were expected to bind to HLA-A33. The peptides were screened for their ability to reactivate memory T lymphocytes from CMV-seropositive HLA-A33 donors by using quantitative real-time RT-PCR. The most promising peptides were then tested for their ability to expand a CD8+ population of HLA-A33 CTLs that produced interferon-γ (IFN-γ) and were cytotoxic to either peptide-loaded or CMV-infected target cells. Results. Sixteen out of 250 peptides were selected using a computer algorithm and peptide stimulation by 8 out of the 16 induced a significant quantity of IFN-γ mRNA transcript from CMV-seropositive HLA-A33 peripheral blood mononuclear cells. All the eight peptides induced consistent T-cell reactivation. One specifically, the peptide pp6591-100 (SVNVHNPTGR), proved to be more active. T cells in vitro sensitized for 2 or 3 weeks with pp6591-100 were cytotoxic to both HLA-A33 peptide-loaded and CMV-infected target cells. Conclusions. CMV pp65 91-100 is a new HLA-A33-restricted peptide that broadens the list of antigenic determinants that can be used for CMV adoptive immunotherapy.",
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T1 - Identification of HLA-A33-restricted CMV pp65 epitopes as common targets for CD8+ CMV-specific cytotoxic T lymphocytes

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AU - Bettinotti, Maria

AU - Caruccio, Lorraine

AU - Nagorsen, Dirk

AU - Stroncek, David

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N2 - Objective. To identify an immunodominant HLA-A33-restricted epitope within the CMV matrix phosphoprotein 65 (pp65) that could be used to elicit CMV-specific CTLs. Methods. A computer algorithm was used to identify pp65 peptides that were expected to bind to HLA-A33. The peptides were screened for their ability to reactivate memory T lymphocytes from CMV-seropositive HLA-A33 donors by using quantitative real-time RT-PCR. The most promising peptides were then tested for their ability to expand a CD8+ population of HLA-A33 CTLs that produced interferon-γ (IFN-γ) and were cytotoxic to either peptide-loaded or CMV-infected target cells. Results. Sixteen out of 250 peptides were selected using a computer algorithm and peptide stimulation by 8 out of the 16 induced a significant quantity of IFN-γ mRNA transcript from CMV-seropositive HLA-A33 peripheral blood mononuclear cells. All the eight peptides induced consistent T-cell reactivation. One specifically, the peptide pp6591-100 (SVNVHNPTGR), proved to be more active. T cells in vitro sensitized for 2 or 3 weeks with pp6591-100 were cytotoxic to both HLA-A33 peptide-loaded and CMV-infected target cells. Conclusions. CMV pp65 91-100 is a new HLA-A33-restricted peptide that broadens the list of antigenic determinants that can be used for CMV adoptive immunotherapy.

AB - Objective. To identify an immunodominant HLA-A33-restricted epitope within the CMV matrix phosphoprotein 65 (pp65) that could be used to elicit CMV-specific CTLs. Methods. A computer algorithm was used to identify pp65 peptides that were expected to bind to HLA-A33. The peptides were screened for their ability to reactivate memory T lymphocytes from CMV-seropositive HLA-A33 donors by using quantitative real-time RT-PCR. The most promising peptides were then tested for their ability to expand a CD8+ population of HLA-A33 CTLs that produced interferon-γ (IFN-γ) and were cytotoxic to either peptide-loaded or CMV-infected target cells. Results. Sixteen out of 250 peptides were selected using a computer algorithm and peptide stimulation by 8 out of the 16 induced a significant quantity of IFN-γ mRNA transcript from CMV-seropositive HLA-A33 peripheral blood mononuclear cells. All the eight peptides induced consistent T-cell reactivation. One specifically, the peptide pp6591-100 (SVNVHNPTGR), proved to be more active. T cells in vitro sensitized for 2 or 3 weeks with pp6591-100 were cytotoxic to both HLA-A33 peptide-loaded and CMV-infected target cells. Conclusions. CMV pp65 91-100 is a new HLA-A33-restricted peptide that broadens the list of antigenic determinants that can be used for CMV adoptive immunotherapy.

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