title: Umbilical cord blood regulatory T-cell expansion and functional effects of tumor necrosis factor receptor family members OX40 and 4-1BB expressed on artificial antigen-presenting cells.
abstract: Previously, we showed that human umbilical cord blood (UCB) regulatory T cells (Tregs) could be expanded approximately 100-fold using anti-CD3/28 monoclonal antibody (mAb)-coated beads to provide T-cell receptor and costimulatory signals. Because Treg numbers from a single UCB unit are limited, we explored the use of cell-based artificial antigen-presenting cells (aAPCs) preloaded with anti-CD3/28 mAbs to achieve higher levels of Treg expansion. Compared with beads, aAPCs had similar expansion properties while significantly increasing transforming growth factor beta (TGF-beta) secretion and the potency of Treg suppressor function. aAPCs modified to coexpress OX40L or 4-1BBL expanded UCB Tregs to a significantly greater extent than bead- or nonmodified aAPC cultures, reaching mean expansion levels exceeding 1250-fold. Despite the high expansion and in contrast to studies using other Treg sources, neither OX40 nor 4-1BB signaling of UCB Tregs reduced in vitro suppression. UCB Tregs expanded with 4-1BBL expressing aAPCs had decreased levels of proapoptotic bim. UCB Tregs expanded with nonmodified or modified aAPCs versus beads resulted in higher survival associated with increased Treg persistence in a xeno-geneic graft-versus-host disease lethality model. These data offer a novel approach for UCB Treg expansion using aAPCs, including those coexpressing OX40L or 4-1BBL.
authors.affiliation: University of Minnesota Cancer Center and Department of Pediatrics, Division of Bone, Blood and Marrow Transplantation, Minneapolis, USA. hippe002@umn.edu
authors.forename: Keli L
authors.initials: KL
authors.lastname: Hippen
authors.forename: Paul
authors.initials: P
authors.lastname: Harker-Murray
authors.forename: Stephen B
authors.initials: SB
authors.lastname: Porter
authors.forename: Sarah C
authors.initials: SC
authors.lastname: Merkel
authors.forename: Aryel
authors.initials: A
authors.lastname: Londer
authors.forename: Dawn K
authors.initials: DK
authors.lastname: Taylor
authors.forename: Megan
authors.initials: M
authors.lastname: Bina
authors.forename: Angela
authors.initials: A
authors.lastname: Panoskaltsis-Mortari
authors.forename: Pablo
authors.initials: P
authors.lastname: Rubinstein
authors.forename: Nico
authors.initials: N
authors.lastname: Van Rooijen
authors.forename: Tatiana N
authors.initials: TN
authors.lastname: Golovina
authors.forename: Megan M
authors.initials: MM
authors.lastname: Suhoski
authors.forename: Jeffrey S
authors.initials: JS
authors.lastname: Miller
authors.forename: John E
authors.initials: JE
authors.lastname: Wagner
authors.forename: Carl H
authors.initials: CH
authors.lastname: June
authors.forename: James L
authors.initials: JL
authors.lastname: Riley
authors.forename: Bruce R
authors.initials: BR
authors.lastname: Blazar
mesh_terms: D053330:4-1BB Ligand / Q000378:metabolism; D000818:Animals; D000938:Antigen-Presenting Cells / Q000187:drug effects / Q000276:immunology*; D051017:Apoptosis Regulatory Proteins / Q000378:metabolism; D000072224:Bcl-2-Like Protein 11; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D005312:Fetal Blood / Q000166:cytology* / Q000187:drug effects; D006086:Graft vs Host Disease / Q000276:immunology; D006801:Humans; D008565:Membrane Proteins / Q000378:metabolism; D051379:Mice; D008863:Microspheres; D011518:Proto-Oncogene Proteins / Q000378:metabolism; D053262:Receptors, OX40 / Q000276:immunology*; D020123:Sirolimus / Q000494:pharmacology; D016019:Survival Analysis; D050378:T-Lymphocytes, Regulatory / Q000166:cytology* / Q000187:drug effects / Q000276:immunology; D053261:Tumor Necrosis Factor Receptor Superfamily, Member 9 / Q000276:immunology*
chemical_list: D053330:4-1BB Ligand; D051017:Apoptosis Regulatory Proteins; C000606775:BCL2L11 protein, human; D000072224:Bcl-2-Like Protein 11; C000606776:Bcl2l11 protein, mouse; D008565:Membrane Proteins; D011518:Proto-Oncogene Proteins; D053262:Receptors, OX40; D053261:Tumor Necrosis Factor Receptor Superfamily, Member 9; D020123:Sirolimus