title: A panel of artificial APCs expressing prevalent HLA alleles permits generation of cytotoxic T cells specific for both dominant and subdominant viral epitopes for adoptive therapy.
abstract: Adoptive transfer of virus-specific T cells can treat infections complicating allogeneic hematopoietic cell transplants. However, autologous APCs are often limited in supply. In this study, we describe a panel of artificial APCs (AAPCs) consisting of murine 3T3 cells transduced to express human B7.1, ICAM-1, and LFA-3 that each stably express one of a series of six common HLA class I alleles. In comparative analyses, T cells sensitized with AAPCs expressing a shared HLA allele or autologous APCs loaded with a pool of 15-mer spanning the sequence of CMVpp65 produced similar yields of HLA-restricted CMVpp65-specific T cells; significantly higher yields could be achieved by sensitization with AAPCs transduced to express the CMVpp65 protein. T cells generated were CD8(+), IFN-gamma(+), and exhibited HLA-restricted CMVpp65-specific cytotoxicity. T cells sensitized with either peptide-loaded or transduced AAPCs recognized epitopes presented by each HLA allele known to be immunogenic in humans. Sensitization with AAPCs also permitted expansion of IFN-gamma(+) cytotoxic effector cells against subdominant epitopes that were either absent or in low frequencies in T cells sensitized with autologous APCs. This replenishable panel of AAPCs can be used for immediate sensitization and expansion of virus-specific T cells of desired HLA restriction for adoptive immunotherapy. It may be of particular value for recipients of transplants from HLA-disparate donors.
authors.affiliation: Department of Pediatrics, Memorial Sloan-Kettering Cancer Center, New York, NY 10021, USA.
authors.forename: Aisha N
authors.initials: AN
authors.lastname: Hasan
authors.forename: Wouter J
authors.initials: WJ
authors.lastname: Kollen
authors.forename: Deepa
authors.initials: D
authors.lastname: Trivedi
authors.forename: Annamalai
authors.initials: A
authors.lastname: Selvakumar
authors.forename: Bo
authors.initials: B
authors.lastname: Dupont
authors.forename: Michel
authors.initials: M
authors.lastname: Sadelain
authors.forename: Richard J
authors.initials: RJ
authors.lastname: O'Reilly
mesh_terms: D000483:Alleles; D000818:Animals; D002478:Cells, Cultured; D003587:Cytomegalovirus / Q000276:immunology*; D018984:Epitopes, T-Lymphocyte / Q000276:immunology; D015234:HLA-A Antigens / Q000235:genetics; D015789:HLA-A2 Antigen; D059871:HLA-A24 Antigen; D015791:HLA-A3 Antigen; D015235:HLA-B Antigens / Q000235:genetics; D015793:HLA-B7 Antigen; D015795:HLA-B8 Antigen; D015395:Histocompatibility Antigens Class I / Q000096:biosynthesis / Q000235:genetics*; D006801:Humans; D016056:Immunodominant Epitopes / Q000008:administration & dosage / Q000276:immunology*; D016219:Immunotherapy, Adoptive; D008213:Lymphocyte Activation / Q000276:immunology*; D051379:Mice; D041681:NIH 3T3 Cells; D010750:Phosphoproteins / Q000008:administration & dosage / Q000276:immunology*; D016176:T-Lymphocyte Subsets / Q000276:immunology / Q000637:transplantation / Q000821:virology; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology / Q000637:transplantation* / Q000821:virology; D014763:Viral Matrix Proteins / Q000008:administration & dosage / Q000276:immunology*
chemical_list: D018984:Epitopes, T-Lymphocyte; D015234:HLA-A Antigens; C435939:HLA-A*02:01 antigen; C442141:HLA-A*03:01 antigen; C438506:HLA-A*24:02 antigen; D015789:HLA-A2 Antigen; D059871:HLA-A24 Antigen; D015791:HLA-A3 Antigen; D015235:HLA-B Antigens; C477444:HLA-B*07:02 antigen; C509422:HLA-B*08:01 antigen; D015793:HLA-B7 Antigen; D015795:HLA-B8 Antigen; D015395:Histocompatibility Antigens Class I; D016056:Immunodominant Epitopes; D010750:Phosphoproteins; D014763:Viral Matrix Proteins; C065418:cytomegalovirus matrix protein 65kDa