title: 4-1BB and CD28 signaling plays a synergistic role in redirecting umbilical cord blood T cells against B-cell malignancies.
abstract: Umbilical cord blood (UCB) T cells can be redirected to kill leukemia and lymphoma cells by engineering with a single-chain chimeric antigen receptor (CAR) and thus may have general applications in adoptive cell therapy. However, the role of costimulatory molecules in UCB T-cell activation and effector functions in context with CAR remains elusive. To investigate the effect of costimulatory molecules (4-1BB and CD28) on UCB T cells, we transduced UCB T cells with lentiviral vectors expressing Green Fluorescent Protein (GFP) and CAR for CD19 containing an intracellular domain of the CD3zeta chain and either a 4-1BB (UCB-19BBzeta) or a CD28 intracellular domain (UCB-1928zeta), both (UCB-1928BBzeta), or neither (UCB-19zeta). We found that UCB-19BBzeta and UCB-28BBzeta T cells exhibited more cytotoxicity to CD19(+) leukemia and lymphoma cell lines than UCB-19zeta and UCB-1928zeta, although differences in secretion of interleukin-2 and interferon-gamma by these T cells were not evident. In vivo adoptive transfer of these T cells into intraperitoneal tumor-bearing mice demonstrated that UCB-19BBzeta and UCB-1928BBzeta T cells mounted the most potent antitumor response. The mice adoptively transferred with UCB-1928BBzeta cells survived longer than the mice with UCB-19BBzeta. Moreover, UCB-1928BBzeta T cells mounted a more robust antitumor response than UCB-19BBzeta in a systemic tumor model. Our data suggest a synergistic role of 4-1BB and CD28 costimulation in engineering antileukemia UCB effector cells and implicate a design for redirected UCB T-cell therapy for refractory leukemia.
authors.affiliation: Graduate Program in Microbial Engineering, University of Minnesota , Minneapolis, MN 55455, USA.
authors.forename: Syam
authors.initials: S
authors.lastname: Tammana
authors.forename: Xin
authors.initials: X
authors.lastname: Huang
authors.forename: Marianna
authors.initials: M
authors.lastname: Wong
authors.forename: Michael C
authors.initials: MC
authors.lastname: Milone
authors.forename: Linan
authors.initials: L
authors.lastname: Ma
authors.forename: Bruce L
authors.initials: BL
authors.lastname: Levine
authors.forename: Carl H
authors.initials: CH
authors.lastname: June
authors.forename: John E
authors.initials: JE
authors.lastname: Wagner
authors.forename: Bruce R
authors.initials: BR
authors.lastname: Blazar
authors.forename: Xianzheng
authors.initials: X
authors.lastname: Zhou
mesh_terms: D000818:Animals; D018941:Antigens, CD19 / Q000378:metabolism; D001402:B-Lymphocytes / Q000276:immunology / Q000473:pathology*; D018106:CD28 Antigens / Q000378:metabolism*; D045744:Cell Line, Tumor; D003602:Cytotoxicity, Immunologic; D005312:Fetal Blood / Q000166:cytology*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006801:Humans; D016130:Immunophenotyping; D007371:Interferon-gamma / Q000096:biosynthesis; D007376:Interleukin-2 / Q000096:biosynthesis; D016086:Lentivirus / Q000235:genetics; D007938:Leukemia / Q000276:immunology / Q000473:pathology / Q000628:therapy*; D008223:Lymphoma / Q000276:immunology / Q000473:pathology; D051379:Mice; D016513:Mice, SCID; D011994:Recombinant Proteins / Q000378:metabolism; D015398:Signal Transduction; D013601:T-Lymphocytes / Q000166:cytology / Q000276:immunology*; D014161:Transduction, Genetic; D053261:Tumor Necrosis Factor Receptor Superfamily, Member 9 / Q000378:metabolism*
chemical_list: D018941:Antigens, CD19; D018106:CD28 Antigens; D007376:Interleukin-2; D011994:Recombinant Proteins; D053261:Tumor Necrosis Factor Receptor Superfamily, Member 9; D049452:Green Fluorescent Proteins; D007371:Interferon-gamma