Characterization, biology, and expansion of regulatory T cells in the Cynomolgus macaque for preclinical studies

Autor: Sigal Kofman, Jonah Zitsman, Megan Sykes, Jeffrey Stern, David C. Woodland, Siu Hong Ho, Leo Buhler, Adam Griesemer, Paula Alonso-Guallart, Raimon Duran-Struuck, Hugo P. Sondermeijer
Přispěvatelé: RS: CARIM - R3.08 - Regenerative and reconstructive medicine for vascular disease, Promovendi CD, Fysiologie, RS: Carim - V03 Regenerative and reconstructive medicine vascular disease
Rok vydání: 2019
Předmět:
graft survival
basic (laboratory) research
cellular transplantation (nonislet)
030230 surgery
THERAPY
T-Lymphocytes
Regulatory
0302 clinical medicine
Immunology and Allergy
Pharmacology (medical)
science
immunobiology
Cells
Cultured
immunosuppression
tolerance
INFUSION
FOXP3
Forkhead Transcription Factors
hemic and immune systems
animal models
chimerism
SURVIVAL
ALLOGRAFT TOLERANCE
EXPRESSION
CD58
BETA
Antigen-Presenting Cells
nonhuman primate
chemical and pharmacologic phenomena
Biology
HEMATOPOIETIC CHIMERISM
Peripheral blood mononuclear cell
Article
03 medical and health sciences
Artificial antigen presenting cells
Animals
Interleukin-7 receptor
Transplantation
immune modulation
immune regulation
IN-VITRO
DNA Methylation
In vitro
Macaca fascicularis
translational research
Immunology
Leukocytes
Mononuclear
HAPLOTYPES
CD8
CD80
Zdroj: Am J Transplant
American Journal of Transplantation, 19(8), 2186-2198. Wiley
ISSN: 1600-6135
DOI: 10.1111/ajt.15313
Popis: 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 (aAPCs) expressing human CD80, CD58 and CD32 were used throughout the culture. When donor peripheral blood mononuclear cells (PBMCs) were used as the single source of APCs followed by aAPCs, Tregs expanded 2000-fold. Tregs from all protocols suppressed the proliferation of anti-CD2CD3CD28 bead-stimulated autologous PBMCs 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, CD8 contamination was observed and resolved by resorting. Specificity studies showed greater suppression of stimulation by anti-CD2CD3CD28 beads of PBMCs 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 CD45RA or CD127 expression. In summary, we have characterized MCM Tregs and developed four Treg expansion protocols that can be used for preclinical applications.
Databáze: OpenAIRE
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