Comprehensive epitope mapping of the Epstein-Barr virus latent membrane protein-2 in normal, non tumor-bearing individuals

Maurizio Provenzano, Silvia Selleri, Ping Jin, Ena Wang, Rosemary Werden, Stephanie Slezak, Sharon D. Adams, Monica C. Panelli, Susan F. Leitman, David F. Stroncek, Francesco M. Marincola

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

Latent membrane protein (LMP)-2 is one of the Epstein-Barr virus (EBV)-encoded proteins consistently expressed by nasopharyngeal carcinoma (NPC). EBV-transformed lymphoblastoid cell lines (LCL) have been used in patients with NPC to induce LMP-2-recognizing T cell lines which have been in turn utilized for protein-wide mapping of T cell epitopes. However, comprehensive mapping of naturally recognized LMP-2 epitopes in non tumor-bearing individuals has not been reported. Here, we applied a low sensitivity epitope-defining technique for the identification of LMP-2 CTL responses detectable ex vivo in EBV-experienced individuals. This screening tool has been previously validated by analyzing memory CTL responses to Flu, cytomegalovirus (CMV), and the melanoma associated antigen gp100/Mel17. Peripheral blood monocytes (PBMC) from ten Caucasian and ten Chinese individuals were stimulated ex vivo with pools of nonamer (9-mer) peptides overlapping in a stepwise fashion each single amino acid of the LMP-2 sequence. No obvious differences were observed between the immune response of the two ethnic groups save for those related to the divergence in the ethnic prevalence of HLA haplotypes. Several novel and known LMP-2 epitopes were identified. Reactivity toward at least one LMP-2 epitope was detected in 18 of the 20 donors but no prevalent human leukocyte antigen (HLA)/epitope combination was observed confirming that LMP-2 reactivity in the context of common HLA alleles is more pleiotropic than that of FLU and CMV. We believe that the usefulness of these epitopes occurring naturally in non-cancer bearing patients as reagents for the immunization of patients with early or advanced stage NPC deserves further evaluation.

Original languageEnglish
Pages (from-to)1047-1063
Number of pages17
JournalCancer Immunology, Immunotherapy
Volume56
Issue number7
DOIs
Publication statusPublished - Jul 2007
Externally publishedYes

Fingerprint

Epitope Mapping
Human Herpesvirus 4
Membrane Proteins
Epitopes
Neoplasms
HLA Antigens
Cytomegalovirus
gp100 Melanoma Antigen
Transformed Cell Line
T-Lymphocyte Epitopes
Ethnic Groups
Haplotypes
Monocytes
Immunization
Proteins
Alleles
Tissue Donors
T-Lymphocytes
Amino Acids
Cell Line

Keywords

  • Antigens/peptides/epitopes
  • Cell activation
  • T cells
  • Tumor immunity

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology
  • Oncology
  • Cancer Research

Cite this

Comprehensive epitope mapping of the Epstein-Barr virus latent membrane protein-2 in normal, non tumor-bearing individuals. / Provenzano, Maurizio; Selleri, Silvia; Jin, Ping; Wang, Ena; Werden, Rosemary; Slezak, Stephanie; Adams, Sharon D.; Panelli, Monica C.; Leitman, Susan F.; Stroncek, David F.; Marincola, Francesco M.

In: Cancer Immunology, Immunotherapy, Vol. 56, No. 7, 07.2007, p. 1047-1063.

Research output: Contribution to journalArticle

Provenzano, M, Selleri, S, Jin, P, Wang, E, Werden, R, Slezak, S, Adams, SD, Panelli, MC, Leitman, SF, Stroncek, DF & Marincola, FM 2007, 'Comprehensive epitope mapping of the Epstein-Barr virus latent membrane protein-2 in normal, non tumor-bearing individuals', Cancer Immunology, Immunotherapy, vol. 56, no. 7, pp. 1047-1063. https://doi.org/10.1007/s00262-006-0246-3
Provenzano, Maurizio ; Selleri, Silvia ; Jin, Ping ; Wang, Ena ; Werden, Rosemary ; Slezak, Stephanie ; Adams, Sharon D. ; Panelli, Monica C. ; Leitman, Susan F. ; Stroncek, David F. ; Marincola, Francesco M. / Comprehensive epitope mapping of the Epstein-Barr virus latent membrane protein-2 in normal, non tumor-bearing individuals. In: Cancer Immunology, Immunotherapy. 2007 ; Vol. 56, No. 7. pp. 1047-1063.
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abstract = "Latent membrane protein (LMP)-2 is one of the Epstein-Barr virus (EBV)-encoded proteins consistently expressed by nasopharyngeal carcinoma (NPC). EBV-transformed lymphoblastoid cell lines (LCL) have been used in patients with NPC to induce LMP-2-recognizing T cell lines which have been in turn utilized for protein-wide mapping of T cell epitopes. However, comprehensive mapping of naturally recognized LMP-2 epitopes in non tumor-bearing individuals has not been reported. Here, we applied a low sensitivity epitope-defining technique for the identification of LMP-2 CTL responses detectable ex vivo in EBV-experienced individuals. This screening tool has been previously validated by analyzing memory CTL responses to Flu, cytomegalovirus (CMV), and the melanoma associated antigen gp100/Mel17. Peripheral blood monocytes (PBMC) from ten Caucasian and ten Chinese individuals were stimulated ex vivo with pools of nonamer (9-mer) peptides overlapping in a stepwise fashion each single amino acid of the LMP-2 sequence. No obvious differences were observed between the immune response of the two ethnic groups save for those related to the divergence in the ethnic prevalence of HLA haplotypes. Several novel and known LMP-2 epitopes were identified. Reactivity toward at least one LMP-2 epitope was detected in 18 of the 20 donors but no prevalent human leukocyte antigen (HLA)/epitope combination was observed confirming that LMP-2 reactivity in the context of common HLA alleles is more pleiotropic than that of FLU and CMV. We believe that the usefulness of these epitopes occurring naturally in non-cancer bearing patients as reagents for the immunization of patients with early or advanced stage NPC deserves further evaluation.",
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AU - Werden, Rosemary

AU - Slezak, Stephanie

AU - Adams, Sharon D.

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AU - Stroncek, David F.

AU - Marincola, Francesco M.

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