title: Sequential monitoring and stability of ex vivo-expanded autologous and nonautologous regulatory T cells following infusion in nonhuman primates.
abstract: Ex vivo-expanded cynomolgus monkey CD4(+)CD25(+)CD127(-) regulatory T cells (Treg) maintained Foxp3 demethylation status at the Treg-specific demethylation region, and potently suppressed T cell proliferation through three rounds of expansion. When carboxyfluorescein succinimidyl ester- or violet proliferation dye 450-labeled autologous (auto) and nonautologous (non-auto)-expanded Treg were infused into monkeys, the number of labeled auto-Treg in peripheral blood declined rapidly during the first week, but persisted at low levels in both normal and anti-thymocyte globulin plus rapamycin-treated (immunosuppressed; IS) animals for at least 3 weeks. By contrast, MHC-mismatched non-auto-Treg could not be detected in normal monkey blood or in blood of two out of the three IS monkeys by day 6 postinfusion. They were also more difficult to detect than auto-Treg in peripheral lymphoid tissue. Both auto- and non-auto-Treg maintained Ki67 expression early after infusion. Sequential monitoring revealed that adoptively transferred auto-Treg maintained similarly high levels of Foxp3 and CD25 and low CD127 compared with endogenous Treg, although Foxp3 staining diminished over time in these nontransplanted recipients. Thus, infused ex vivo-expanded auto-Treg persist longer than MHC-mismatched non-auto-Treg in blood of nonhuman primates and can be detected in secondary lymphoid tissue. Host lymphodepletion and rapamycin administration did not consistently prolong the persistence of non-auto-Treg in these sites.
authors.affiliation: Thomas E. Starzl Transplantation Institute, Department of Surgery, University of Pittsburgh School of Medicine, Pittsburgh, PA.
authors.forename: H
authors.initials: H
authors.lastname: Zhang
authors.forename: H
authors.initials: H
authors.lastname: Guo
authors.forename: L
authors.initials: L
authors.lastname: Lu
authors.forename: A F
authors.initials: AF
authors.lastname: Zahorchak
authors.forename: R W
authors.initials: RW
authors.lastname: Wiseman
authors.forename: G
authors.initials: G
authors.lastname: Raimondi
authors.forename: D K C
authors.initials: DK
authors.lastname: Cooper
authors.forename: M B
authors.initials: MB
authors.lastname: Ezzelarab
authors.forename: A W
authors.initials: AW
authors.lastname: Thomson
mesh_terms: D000818:Animals; D000961:Antilymphocyte Serum / Q000737:chemistry; D051858:Forkhead Transcription Factors / Q000378:metabolism; D006239:Haplotypes; D007166:Immunosuppressive Agents / Q000737:chemistry; D053645:Interleukin-2 Receptor alpha Subunit / Q000378:metabolism*; D053699:Interleukin-7 Receptor alpha Subunit / Q000378:metabolism*; D019394:Ki-67 Antigen / Q000378:metabolism; D008252:Macaca fascicularis; D008285:Major Histocompatibility Complex; D008297:Male; D008745:Methylation; D010641:Phenotype; D020123:Sirolimus / Q000737:chemistry; D050378:T-Lymphocytes, Regulatory / Q000276:immunology*
chemical_list: D000961:Antilymphocyte Serum; D051858:Forkhead Transcription Factors; D007166:Immunosuppressive Agents; D053645:Interleukin-2 Receptor alpha Subunit; D053699:Interleukin-7 Receptor alpha Subunit; D019394:Ki-67 Antigen; D020123:Sirolimus
keywords: Immunosuppressant; T cell biology; animal models: nonhuman primate; basic (laboratory) research / science; immunobiology; immunosuppressant; immunosuppression / immune modulation; mechanistic target of rapamycin (mTOR); polyclonal preparations: rabbit antithymocyte globulin; translational research / science