Optimization of LMP-specific CTL expansion for potential adoptive immunotherapy in NPC patients.
Nasopharyngeal carcinoma (NPC) is Epstein-Barr virus (EBV) positive in all undifferentiated cases, expressing the latency II phenotype of latent membrane proteins (LMPs) 1 and 2, in addition to EBV nuclear antigen (EBNA) 1. Several studies have attempted to treat NPC with EBV-specific cytotoxic T lymphocyte (CTL) with a partial response. To improve this therapy, there is a need to expand CTL targeted to the latency II antigens of EBV, rather than the immunodominant EBV nuclear antigens 3-6 peptides typically expanded by lymphoblastoid cells. In order to maximize the expansion of LMP-specific CTL in vitro for use in adoptive immunotherapy of nasopharyngeal carcinoma patients, we used lymphoblastoid cell lines coated with synthetic peptides corresponding to CTL determinants from the LMP proteins. We investigated several issues pertaining to the expansion of an immunologically weak CTL response, including peptide and interleukin-2 concentration, and screening assays for selecting the optimal peptide for use in expansion of LMP-specific CTL. Although screening of ex vivo peripheral blood mononuclear cells did not prove to be useful in the selection of an LMP peptide for use in CTL cultures, the peptide and interleukin-2 concentrations were critical for the maximum expansion of CTL. Therefore, it is imperative that stimulation protocols are optimized for the expansion of LMP-specific CTL.
Lutzky
Viviana P
VP
EBV Biology Laboratory, Division of Immunology, Australian Centre for Vaccine Development, The Queensland Institute of Medical Research, The Royal Brisbane Hospital, 300 Herston Road, Herston, Brisbane, QLD 4006, Australia. viviana.lutzky@qimr.edu.au
Davis
Joanne E
JE
Crooks
Pauline
P
Corban
Monika
M
Smith
Mark C
MC
Elliott
Michael
M
Morrison
Leanne
L
Cross
Simone
S
Tscharke
David
D
Panizza
Benedict
B
Coman
William
W
Bharadwaj
Mandvi
M
Moss
Denis J
DJ
D000938:Antigen-Presenting Cells / Q000276:immunology* / Q000378:metabolism; D049109:Cell Proliferation; D002478:Cells, Cultured; D019309:Epstein-Barr Virus Nuclear Antigens / Q000276:immunology / Q000378:metabolism; D006680:HLA Antigens / Q000378:metabolism; D004854:Herpesvirus 4, Human / Q000276:immunology*; D006801:Humans; D016056:Immunodominant Epitopes / Q000737:chemistry / Q000276:immunology / Q000378:metabolism; D016219:Immunotherapy, Adoptive; D007371:Interferon-gamma / Q000378:metabolism; D008213:Lymphocyte Activation; D009303:Nasopharyngeal Neoplasms / Q000276:immunology* / Q000378:metabolism / Q000473:pathology / Q000628:therapy* / Q000821:virology; D010446:Peptide Fragments / Q000276:immunology / Q000378:metabolism; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology / Q000378:metabolism* / Q000473:pathology; D014763:Viral Matrix Proteins / Q000276:immunology* / Q000378:metabolism
C037662:EBV-associated membrane antigen, Epstein-Barr virus; D019309:Epstein-Barr Virus Nuclear Antigens; D006680:HLA Antigens; D016056:Immunodominant Epitopes; D010446:Peptide Fragments; D014763:Viral Matrix Proteins; D007371:Interferon-gamma; C417029:EBV-encoded nuclear antigen 1