title: In vitro expanded human CD4+CD25+ regulatory T cells suppress effector T cell proliferation.
abstract: Regulatory T cells (Tregs) have been shown to be critical in the balance between autoimmunity and tolerance and have been implicated in several human autoimmune diseases. However, the small number of Tregs in peripheral blood limits their therapeutic potential. Therefore, we developed a protocol that would allow for the expansion of Tregs while retaining their suppressive activity. We isolated CD4+CD25 hi cells from human peripheral blood and expanded them in vitro in the presence of anti-CD3 and anti-CD28 magnetic Xcyte Dynabeads and high concentrations of exogenous Interleukin (IL)-2. Tregs were effectively expanded up to 200-fold while maintaining surface expression of CD25 and other markers of Tregs: CD62L, HLA-DR, CCR6, and FOXP3. The expanded Tregs suppressed proliferation and cytokine secretion of responder PBMCs in co-cultures stimulated with anti-CD3 or alloantigen. Treg expansion is a critical first step before consideration of Tregs as a therapeutic intervention in patients with autoimmune or graft-versus-host disease.
authors.affiliation: Department of Medicine, UCSF Diabetes Center, University of California, Box 0540, 513 Parnassus Avenue, San Francisco, CA 94143-0540, USA.
authors.forename: K E
authors.initials: KE
authors.lastname: Earle
authors.forename: Q
authors.initials: Q
authors.lastname: Tang
authors.forename: X
authors.initials: X
authors.lastname: Zhou
authors.forename: W
authors.initials: W
authors.lastname: Liu
authors.forename: S
authors.initials: S
authors.lastname: Zhu
authors.forename: M L
authors.initials: ML
authors.lastname: Bonyhadi
authors.forename: J A
authors.initials: JA
authors.lastname: Bluestone
mesh_terms: D001327:Autoimmune Diseases / Q000628:therapy; D015496:CD4-Positive T-Lymphocytes / Q000166:cytology / Q000276:immunology*; D018929:Cell Culture Techniques; D005434:Flow Cytometry; D051858:Forkhead Transcription Factors; D006684:HLA-DR Antigens / Q000276:immunology; D006801:Humans; D018189:Immunomagnetic Separation; D016130:Immunophenotyping; D007167:Immunotherapy / Q000379:methods; D007376:Interleukin-2 / Q000276:immunology; D019041:L-Selectin / Q000276:immunology; D008213:Lymphocyte Activation / Q000276:immunology; D054399:Receptors, CCR6; D019707:Receptors, Chemokine / Q000276:immunology; D015375:Receptors, Interleukin-2 / Q000276:immunology*; D012097:Repressor Proteins
chemical_list: C106311:CCR6 protein, human; C486626:FOXP1 protein, human; D051858:Forkhead Transcription Factors; D006684:HLA-DR Antigens; D007376:Interleukin-2; D054399:Receptors, CCR6; D019707:Receptors, Chemokine; D015375:Receptors, Interleukin-2; D012097:Repressor Proteins; D019041:L-Selectin