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Characterization, biology, and expansion of regulatory T cells in the Cynomolgus macaque for preclinical studies
Author(s) -
AlonsoGuallart Paula,
Zitsman Jonah S.,
Stern Jeffrey,
Kofman Sigal B.,
Woodland David,
Ho SiuHong,
Sondermeijer Hugo P.,
Bühler Leo,
Griesemer Adam,
Sykes Megan,
DuranStruuck Raimon
Publication year - 2019
Publication title -
american journal of transplantation
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.89
H-Index - 188
eISSN - 1600-6143
pISSN - 1600-6135
DOI - 10.1111/ajt.15313
Subject(s) - foxp3 , peripheral blood mononuclear cell , cd80 , immunology , interleukin 7 receptor , in vitro , medicine , biology , immune system , cancer research , cytotoxic t cell , il 2 receptor , t cell , cd40 , genetics
Reliable in vitro expansion protocols of regulatory T cells (Tregs) are needed for clinical use. We studied the biology of Mauritian Cynomolgus macaque ( MCM ) Tregs and developed four in vitro Treg expansion protocols for translational studies. Tregs expanded 3000‐fold when artificial antigen presenting cells ( aAPC s) expressing human CD 80, CD 58 and CD 32 were used throughout the culture. When donor peripheral blood mononuclear cells ( PBMC s) were used as the single source of APC s followed by aAPC s, Tregs expanded 2000‐fold. Tregs from all protocols suppressed the proliferation of anti‐ CD 2 CD 3 CD 28 bead‐stimulated autologous PBMC s albeit with different potencies, varying from 1:2‐1:4 Treg: PBMC ratios, up to >1:32. Reculture of cryopreserved Tregs permitted reexpansion with improved suppressive activity. Occasionally, CD 8 contamination was observed and resolved by resorting. Specificity studies showed greater suppression of stimulation by anti‐ CD 2 CD 3 CD 28 beads of PBMC s from the same donor used for stimulation during the Treg cultures and of autologous cells than of third‐party PBMC responders. Similar to humans, the Treg–specific demethylated region ( TSDR ) within the Foxp3 locus correlated with suppressive activity and expression of Foxp3. Contrary to humans, FoxP3 expression did not correlate with CD 45 RA or CD 127 expression. In summary, we have characterized MCM Tregs and developed four Treg expansion protocols that can be used for preclinical applications.