title: 4-1BB is superior to CD28 costimulation for generating CD8+ cytotoxic lymphocytes for adoptive immunotherapy.
abstract: Artificial APCs (aAPCs) genetically modified to express selective costimulatory molecules provide a reproducible, cost-effective, and convenient method for polyclonal and Ag-specific expansion of human T cells for adoptive immunotherapy. Among the variety of aAPCs that have been studied, acellular beads expressing anti-CD3/anti-CD28 efficiently expand CD4+ cells, but not CD8+ T cells. Cell-based aAPCs can effectively expand cytolytic CD8+ cells, but optimal costimulatory signals have not been defined. 4-1BB, a costimulatory molecule expressed by a minority of resting CD8+ T cells, is transiently up-regulated by all CD8+ T cells following activation. We compared expansion of human cytolytic CD8+ T cells using cell-based aAPCs providing costimulation via 4-1BB vs CD28. Whereas anti-CD3/anti-CD28 aAPCs mostly expand naive cells, anti-CD3/4-1BBL aAPCs preferentially expand memory cells, resulting in superior enrichment of Ag-reactive T cells which recognize previously primed Ags and efficient expansion of electronically sorted CD8+ populations reactive toward viral or self-Ags. Using HLA-A2-Fc fusion proteins linked to 4-1BBL aAPCs, 3-log expansion of Ag-specific CD8+ CTL was induced over 14 days, whereas similar Ag-specific CD8+ T cell expansion did not occur using HLA-A2-Fc/anti-CD28 aAPCs. Furthermore, when compared with cytolytic T cells expanded using CD28 costimulation, CTL expanded using 4-1BB costimulation mediate enhanced cytolytic capacity due, in part, to NKG2D up-regulation. These results demonstrate that 4-1BB costimulation is essential for expanding memory CD8+ T cells ex vivo and is superior to CD28 costimulation for generating Ag-specific products for adoptive cell therapy.
authors: Pediatric Oncology Branch, National Cancer Institute, Bethesda, MD 20892, USA.
authors: Hua
authors: H
authors: Zhang
authors: Kristen M
authors: KM
authors: Snyder
authors: Megan M
authors: MM
authors: Suhoski
authors: Marcela V
authors: MV
authors: Maus
authors: Veena
authors: V
authors: Kapoor
authors: Carl H
authors: CH
authors: June
authors: Crystal L
authors: CL
authors: Mackall
mesh_terms: D053330:4-1BB Ligand / Q000378:metabolism*; D000938:Antigen-Presenting Cells / Q000378:metabolism; D000951:Antigens, Neoplasm / Q000378:metabolism; D018106:CD28 Antigens / Q000276:immunology* / Q000378:metabolism*; D017252:CD3 Complex / Q000276:immunology; D049109:Cell Proliferation; D002478:Cells, Cultured; D006649:Histocompatibility Antigens / Q000276:immunology; D006801:Humans; D007156:Immunologic Memory / Q000276:immunology; D016219:Immunotherapy, Adoptive; D058965:MART-1 Antigen; D055655:NK Cell Lectin-Like Receptor Subfamily K; D009363:Neoplasm Proteins / Q000378:metabolism; D011948:Receptors, Antigen, T-Cell / Q000276:immunology; D011961:Receptors, Fc / Q000276:immunology; D011971:Receptors, Immunologic / Q000378:metabolism; D055607:Receptors, Natural Killer Cell; D015398:Signal Transduction; D013602:T-Lymphocytes, Cytotoxic / Q000166:cytology / Q000276:immunology* / Q000378:metabolism*
chemical_list: D053330:4-1BB Ligand; D000951:Antigens, Neoplasm; D018106:CD28 Antigens; D017252:CD3 Complex; D006649:Histocompatibility Antigens; C527085:KLRK1 protein, human; D058965:MART-1 Antigen; C548667:MLANA protein, human; D055655:NK Cell Lectin-Like Receptor Subfamily K; D009363:Neoplasm Proteins; D011948:Receptors, Antigen, T-Cell; D011961:Receptors, Fc; D011971:Receptors, Immunologic; D055607:Receptors, Natural Killer Cell