title: A modified peptide stimulation method for efficient amplification of cytomegalovirus (CMV)-specific CTLs.
abstract: CMV-specific immunity is essential for control of human cytomegalovirus (HCMV) infection. Stem cell transplantation is used widely in the management of a range of diseases of the hemopoietic system. Patients are immunosuppressed profoundly in the early posttransplant period, and reactivation of cytomegalovirus (CMV) remains a significant cause of morbidity and mortality. Adoptive transfer of CMV-specific CD8+ T cell clones has been shown to reduce the rate of viral reactivation; however, the ex vivo production of cells for adoptive transfer is labor intensive and expensive. We report here a modified peptide stimulation method using CMV-specific epitope peptides to stimulate PBMCs for generation of CMV-specific CTLs. This method permits efficient amplification of CMV-specific CTLs and provides a large number of cells for FACS analysis from a single blood sample. Significantly, it achieves high frequencies of tetramer staining of CD8+ T cells allowing the data of different individuals to be easily compared and sequentially evaluated. Thus, this approach expands and selects HLA-restricted CMV-pp65-reactive T-cell lines of high specificity for potential adoptive immunotherapy.
authors.affiliation: Institute of Molecular Immunology, School of Biotechnology, Southern Medical University, Guangzhou, China.
authors.forename: Guang Ping
authors.initials: GP
authors.lastname: Ruan
authors.forename: Li
authors.initials: L
authors.lastname: Ma
authors.forename: Qian
authors.initials: Q
authors.lastname: Wen
authors.forename: Wei
authors.initials: W
authors.lastname: Luo
authors.forename: Ming Qian
authors.initials: MQ
authors.lastname: Zhou
authors.forename: Xiao Ning
authors.initials: XN
authors.lastname: Wang
mesh_terms: D003587:Cytomegalovirus / Q000276:immunology*; D003586:Cytomegalovirus Infections / Q000276:immunology* / Q000821:virology; D018984:Epitopes, T-Lymphocyte / Q000276:immunology* / Q000378:metabolism; D015789:HLA-A2 Antigen / Q000276:immunology* / Q000378:metabolism; D006801:Humans; D016219:Immunotherapy, Adoptive; D007963:Leukocytes, Mononuclear / Q000276:immunology / Q000378:metabolism; D009842:Oligopeptides / Q000138:chemical synthesis / Q000276:immunology*; D010750:Phosphoproteins / Q000276:immunology*; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology* / Q000378:metabolism; D014763:Viral Matrix Proteins / Q000276:immunology*
chemical_list: D018984:Epitopes, T-Lymphocyte; D015789:HLA-A2 Antigen; D009842:Oligopeptides; D010750:Phosphoproteins; D014763:Viral Matrix Proteins; C065418:cytomegalovirus matrix protein 65kDa