Premium
Induction of porcine‐specific regulatory T cells with high specificity and expression of IL ‐10 and TGF ‐β1 using baboon‐derived tolerogenic dendritic cells
Author(s) -
Li Mingqian,
Eckl Judith,
Abicht JanMichael,
Mayr Tanja,
Reichart Bruno,
Schendel Dolores J.,
Pohla Heike
Publication year - 2017
Publication title -
xenotransplantation
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.052
H-Index - 61
eISSN - 1399-3089
pISSN - 0908-665X
DOI - 10.1111/xen.12355
Subject(s) - baboon , xenotransplantation , biology , immune system , immune tolerance , dendritic cell , immunology , peripheral blood mononuclear cell , t cell , cytokine , in vivo , microbiology and biotechnology , in vitro , transplantation , medicine , biochemistry , surgery , endocrinology
Background Regulatory T cells (Treg) play an important role in maintenance of homeostasis in vivo. Treg application to alleviate allo‐organ rejection is being studied extensively. However, natural Treg ( nT reg) expansion in vitro is laborious and expensive. Antigen‐specific Treg are more effective and require lower cell numbers than use of nT reg for immune control. The baboon, as a non‐human primate experimental animal model, is widely used in xenotransplantation research. An effective method to generate baboon xeno‐specific Treg would benefit research on immune tolerance in xenotransplantation using this model system. Method Baboon tolerogenic dendritic cells (tol DC ) were generated in 3 days from monocytes isolated from baboon peripheral blood mononuclear cells in medium supplemented with anti‐inflammatory cytokines. After loading with porcine‐specific ( PS ) in vitro‐transcribed RNA (ivt RNA ), tol DC were used to induce CD 4 + T cells to become porcine‐specific Treg ( PST reg) in cocultures supplemented with IL ‐2 and rapamycin for 10 days. Anti‐inflammatory and inflammatory cytokine expression was evaluated at the mRNA and protein levels in both baboon tol DC and PST reg. Functional assays, suppression of activation markers on porcine‐specific effector T cells ( PST eff) and inhibition of PST eff proliferation, were used to test PST reg specificity. Results Tol DC generated with this method exhibited a tolerogenic phenotype, expressed CCR 7 and produced high levels of IL ‐10 and TGF ‐β1, whereas IL ‐12p40 and IFN ‐γ were not expressed. PST reg were successfully generated in cocultures of CD 4 + T cells and PS ivt RNA ‐loaded tol DC . They exhibited a CD 3 + CD 4 + CD 25 + CD 127 low/− CD 45 RA low Foxp3 + phenotype and were characterized by high expression of IL ‐10 and TGF ‐β1 mRNA and protein. They showed upregulated expression of EBI 3 and GARP mRNA . PST reg exhibited highly suppressive effects toward PST eff, secreting high amounts of IL ‐10 and TGF ‐β1 cytokine upon interaction with PST eff and suppressing IFN ‐γ expression on PST eff. Conclusion In this study, a fast 3‐day method to generate baboon‐derived tol DC is provided that allows subsequent induction of PST reg displaying high porcine‐antigen specificity and expression of IL ‐10 and TGF ‐β1. Porcine‐specific baboon Treg can be used in porcine solid organ or cell xenotransplantation studies through adoptive cell transfer into host baboons.