A comparison of gene transfer and antigen-loaded dendritic cells for the generation of CD4 + and CD8 + cytomegalovirus-specific T cells in HLA-A2 + and HLA-A2 − donors

• Published in: Biology of blood and marrow transplantation Vol. 10; no. 11; pp. 761 - 771
• Main Authors: Foster, Aaron E., Bradstock, Kenneth F., Sili, Uluhan, Marangolo, Marina, Rooney, Cliona M., Gottlieb, David J.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.11.2004
• Subjects: Antigens; Blood Donors; CD4-Positive T-Lymphocytes - immunology; CD8-Positive T-Lymphocytes - immunology; Cells, Cultured; CTL; Cytomegalovirus - genetics; Cytomegalovirus - immunology; Dendritic Cells - immunology; Epitopes; HLA-A2 Antigen - immunology; Human; Humans; Peptides; Phosphoproteins - genetics; Phosphoproteins - immunology; Transduction, Genetic; Transplantation; Viral; Viral Matrix Proteins - genetics; Viral Matrix Proteins - immunology; Human; Antigens; Peptides; Viral; CTL; Transplantation; Epitopes
• ISSN: 1083-8791; 1523-6536
• DOI: 10.1016/j.bbmt.2004.05.011

Dendritic cells have been used effectively to select for human cytomegalovirus (CMV)-specific T cells for immunotherapy applications. The ability to process and present relevant major histocompatibility complex class I and II peptides to T cells makes them ideal for selecting CD4 + and CD8 + T cells regardless of HLA tissue type. This study compared the generation of CMV-specific T cells by using dendritic cells loaded with either CMV pp65 (495–503) peptide or CMV lysate or transduced with adenovirus encoding the pp65 gene (Ad5pp65GFP) for the generation of CD4 + and CD8 + CMV-specific T cells in HLA-A2 + and HLA-A2 − donors. In HLA-A2 + donors, CD8 + tetramer + T cells increased with all antigens but were greatest in peptide- and Ad5pp65GFP-stimulated T cells. The CD4 +/CD8 + ratio in the stimulated T-cell cultures proved to be dependent on the antigen used. CMV lysate-stimulated cells were primarily CD4 +, whereas peptide- and Ad5pp65GFP-stimulated cultures were mostly CD8 +. Analysis of cells from lysate-stimulated or gene-transduced-stimulated cultures showed expansion of CMV-specific CD4 + T cells, indicating that major histocompatibility complex class II peptides were present in both antigens. Furthermore, CMV-specific T cells were generated from HLA-A2 − donors by using Ad5pp65GFP transduction or CMV lysate stimulation and were able to recognize a pp65 peptide restricted to the HLA-B35 allele. These data indicate that either CMV lysate or adenovirus encoding CMV antigenic genes may be useful for the generation of both CD4 + and CD8 + CMV-specific T cells in donors irrespective of HLA tissue type and may be applicable to clinical immunotherapy.