Kim, Hyun-Joo Cha, Gil Sun Joo, Ji-Young Lee, Juyoun Kim, Sung-Jo Lee, Jeongae Park, So Youn Choi, Jeomil 2024-01-20T00:31:35Z 2024-01-20T00:31:35Z 2021-09-05 2017-10 2093-2278 https://pubs.kist.re.kr/handle/201004/122222 Purpose: Beyond the limited scope of non-specific polyclonal regulatory T cell (Treg)-based immunotherapy, which depends largely on serendipity, the present study explored a target Treg subset appropriate for the delivery of a novel epitope spreader Pep19 antigen as part of a sophisticated form of immunotherapy with defined antigen specificity that induces immune tolerance. Methods: Human polyclonal CD4(+)CD25(+)CD127(lo)-Tregs (127-Tregs) and naive CD4(+)CD25(+)CD45RA(+) Tregs (45RA-Tregs) were isolated and were stimulated with target peptide 19 (Pep19)-pulsed dendritic cells in a tolerogenic milieu followed by ex vivo expansion. Low-dose interleukin-2 (IL-2) and rapamycin were added to selectively exclude the outgrowth of contaminating effector T cells (Teffs). The following parameters were investigated in the expanded antigen-specific Tregs: the distinct expression of the immunosuppressive Treg marker Foxp3, epigenetic stability (demethylation in the Treg-specific demethylated region), the suppression of Teffs, expression of the homing receptors CD62L/CCR7, and CD95L-mediated apoptosis. The expanded Tregs were adoptively transferred into an NOD/scid/IL2-R gamma(-/-) mouse model of collagen-induced arthritis. Results: Epitope-spreader Pep19 targeting by 45RA-Tregs led to an outstanding in vitro suppressive T cell fate characterized by robust ex vivo expansion, the salient expression of Foxp3, high epigenetic stability, enhanced T cell suppression, modest expression of CD62L/CCR7, and higher resistance to CD95L-mediated apoptosis. After adoptive transfer, the distinct fate of these T cells demonstrated a potent in vivo immunotherapeutic capability, as indicated by the complete elimination of footpad swelling, prolonged survival, minimal histopathological changes, and preferential localization of CD4(+)CD25(+) Tregs at the articular joints in a mechanistic and orchestrated way. Conclusions: We propose human naive CD4(+)CD25(+)CD45RA(+) Tregs and the epitope spreader Pep19 as cellular and molecular targets for a novel antigen-specific Treg-based vaccination against collagen-induced arthritis. English KOREAN ACAD PERIODONTOLOGY HEAT-SHOCK-PROTEIN AUTOIMMUNE-DISEASES FOXP3 EXPRESSION SUBPOPULATION PERIODONTITIS ARTHRITIS CAPACITY ASSAY Targeting the epitope spreader Pep19 by naive human CD45RA(+) regulatory T cells dictates a distinct suppressive T cell fate in a novel form of immunotherapy Article 10.5051/jpis.2017.47.5.292 1 JOURNAL OF PERIODONTAL AND IMPLANT SCIENCE, v.47, no.5, pp.292 - 311 JOURNAL OF PERIODONTAL AND IMPLANT SCIENCE 47 5 292 311 scie scopus kci ART002279733 000416673700004 2-s2.0-85032487707 Dentistry, Oral Surgery & Medicine Dentistry, Oral Surgery & Medicine Article HEAT-SHOCK-PROTEIN AUTOIMMUNE-DISEASES FOXP3 EXPRESSION SUBPOPULATION PERIODONTITIS ARTHRITIS CAPACITY ASSAY Adoptive transfer Autoimmune diseases Heat-shock proteins Immune tolerance Regulatory T-lymphocytes Rheumatoid arthritis