title: Phenotype and functional evaluation of ex vivo generated antigen-specific immune effector cells with potential for therapeutic applications.
abstract: Ex vivo activation and expansion of lymphocytes for adoptive cell therapy has demonstrated great success. To improve safety and therapeutic efficacy, increased antigen specificity and reduced non-specific response of the ex vivo generated immune cells are necessary. Here, using a complete protein-spanning pool of pentadecapeptides of the latent membrane protein 2A (LMP2A) of Epstein-Barr virus (EBV), a weak viral antigen which is associated with EBV lymphoproliferative diseases, we investigated the phenotype and function of immune effector cells generated based on IFN-gamma or CD137 activation marker selection and dendritic cell (DC) activation. These ex vivo prepared immune cells exhibited a donor- and antigen-dependent T cell response; the IFN-gamma-selected immune cells displayed a donor-related CD4- or CD8-dominant T cell phenotype; however, the CD137-enriched cells showed an increased ratio of CD4 T cells. Importantly, the pentadecapeptide antigens accessed both class II and class I MHC antigen processing machineries and effectively activated EBV-specific CD4 and CD8 T cells. Phenotype and kinetic analyses revealed that the IFN-gamma and the CD137 selections enriched more central memory T (Tcm) cells than did the DC-activation approach, and after expansion, the IFN-gamma-selected effector cells showed the highest level of antigen-specificity and effector activities. While all three approaches generated immune cells with comparable antigen-specific activities, the IFN-gamma selection followed by ex vivo expansion produced high quality and quantity of antigen-specific effector cells. Our studies presented the optimal approach for generating therapeutic immune cells with potential for emergency and routine clinical applications.
authors: Department of Molecular Genetics and Microbiology, College of Medicine, University of Florida, Gainesville, FL 32610-0266, USA. hansh72@ufl.edu
authors: Shuhong
authors: S
authors: Han
authors: Yuju
authors: Y
authors: Huang
authors: Yin
authors: Y
authors: Liang
authors: Yuchin
authors: Y
authors: Ho
authors: Yichen
authors: Y
authors: Wang
authors: Lung-Ji
authors: LJ
authors: Chang
mesh_terms: D000941:Antigens / Q000276:immunology*; D018929:Cell Culture Techniques; D002469:Cell Separation / Q000379:methods; D002478:Cells, Cultured; D000939:Epitopes / Q000276:immunology; D006801:Humans; D007156:Immunologic Memory; D016130:Immunophenotyping; D016219:Immunotherapy, Adoptive / Q000379:methods*; D007371:Interferon-gamma / Q000378:metabolism; D008213:Lymphocyte Activation / Q000276:immunology / Q000502:physiology*; D010446:Peptide Fragments / Q000276:immunology; D010641:Phenotype; D013601:T-Lymphocytes / Q000276:immunology* / Q000378:metabolism / Q000637:transplantation; D053261:Tumor Necrosis Factor Receptor Superfamily, Member 9 / Q000378:metabolism; D014763:Viral Matrix Proteins / Q000737:chemistry / Q000276:immunology
chemical_list: D000941:Antigens; C037662:EBV-associated membrane antigen, Epstein-Barr virus; D000939:Epitopes; D010446:Peptide Fragments; D053261:Tumor Necrosis Factor Receptor Superfamily, Member 9; D014763:Viral Matrix Proteins; D007371:Interferon-gamma