PMID- 28673339 OWN - NLM STAT- MEDLINE DCOM- 20180326 LR - 20181113 IS - 1757-6512 (Electronic) IS - 1757-6512 (Linking) VI - 8 IP - 1 DP - 2017 Jul 3 TI - Human limbal fibroblast-like stem cells induce immune-tolerance in autoreactive T lymphocytes from female patients with Hashimoto's thyroiditis. PG - 154 LID - 10.1186/s13287-017-0611-5 [doi] LID - 154 AB - BACKGROUND: Due to their "natural immune privilege" and immunoregulatory properties human fibroblast-like limbal stem cells (f-LSCs) have acquired great interest as a potential tool for achieving immunotolerance. Hashimoto's thyroiditis (HT) is the most common thyroid autoimmune disease and cause of hypothyroidism. To date, conventional hormone replacement therapy and unspecific immunosuppressive regimens cannot provide a definitive cure for HT subjects. We explored the immunosuppressant potential of human f-LSCs on circulating lymphomonocytes (PBMCs) collected from healthy donors and female HT patients. METHODS: We assessed the immunophenotyping of f-LSCs, both untreated and after 48 h of proinflammatory cytokine exposure, by means of quantitative reverse-transcription polymerase chain reaction (qRT-PCR) and flow cytometry. The immunosuppressant effects of f-LSCs on healthy activated PBMCs were investigated in cell-cell contact and transwell settings through cell cycle assay, acridine orange staining, and caspase-3 detection. We also studied T-cell responses and possible Treg conversion by means of flow cytometry. Functional assays were conducted in activated HT lymphocytes cocultured with f-LSCs after carboxyfluorescein succinimidyl ester labeling and intracellular detection of pro- and anti-inflammatory cytokines. RESULTS: The hypo-immunogenicity of the f-LSC population depended on both cell contact and soluble factors produced, as well as the undetectable expression of all those molecules required to fully activate T lymphocytes. Following exposure to Th1 cytokines, f-LSCs augmented expression of programmed death-ligand 1 and 2 (PDL-1 and -2), indoleamine-pyrrole-2,3-dioxygenase (IDO), interleukin (IL)-6, and monocyte chemotactic protein 1 (MCP-1) while maintaining their negative phenotype for major histocompatibility (MHC) class II and costimulatory molecules. During coculture, f-LSCs suppressed up to 40% of proliferation in healthy activated PBMCs, arrested them in the G0/G1 cell cycle phase without inducing apoptosis cascade, inverted the CD4/CD8 ratio, and promoted sustained expression of the immunomodulator marker CD69. Under coculture conditions the Th imbalance of autoreactive T cells from female HT patients was fully restored. CONCLUSIONS: Our study describes an in vitro coculture system able to prevent inappropriate activation of autoreactive T lymphocytes of female HT patients and to generate a tolerogenic environment even in an inflammatory background. Further investigations are necessary to establish whether this stem cell-based therapy approach in HT could avoid lifetime hormone replacement therapy by inducing T-cell education. FAU - Coppola, Antonina AU - Coppola A AD - Laboratory of Regenerative Medicine, Section of Endocrinology, Diabetology and Metabolism, Di.Bi.M.I.S., University of Palermo, Piazza delle Cliniche 2, 90127, Palermo, Italy. AD - ATeN (Advanced Technologies Network Center), University of Palermo, Palermo, Italy. FAU - Tomasello, Laura AU - Tomasello L AD - Laboratory of Regenerative Medicine, Section of Endocrinology, Diabetology and Metabolism, Di.Bi.M.I.S., University of Palermo, Piazza delle Cliniche 2, 90127, Palermo, Italy. AD - ATeN (Advanced Technologies Network Center), University of Palermo, Palermo, Italy. FAU - Pitrone, Maria AU - Pitrone M AD - Laboratory of Regenerative Medicine, Section of Endocrinology, Diabetology and Metabolism, Di.Bi.M.I.S., University of Palermo, Piazza delle Cliniche 2, 90127, Palermo, Italy. AD - ATeN (Advanced Technologies Network Center), University of Palermo, Palermo, Italy. FAU - Cillino, Salvatore AU - Cillino S AD - Department of Ophthalmology, University of Palermo, Palermo, Italy. FAU - Richiusa, Pierina AU - Richiusa P AD - Laboratory of Regenerative Medicine, Section of Endocrinology, Diabetology and Metabolism, Di.Bi.M.I.S., University of Palermo, Piazza delle Cliniche 2, 90127, Palermo, Italy. FAU - Pizzolanti, Giuseppe AU - Pizzolanti G AD - Laboratory of Regenerative Medicine, Section of Endocrinology, Diabetology and Metabolism, Di.Bi.M.I.S., University of Palermo, Piazza delle Cliniche 2, 90127, Palermo, Italy. giuseppe.pizzolanti@unipa.it. AD - ATeN (Advanced Technologies Network Center), University of Palermo, Palermo, Italy. giuseppe.pizzolanti@unipa.it. FAU - Giordano, Carla AU - Giordano C AD - Laboratory of Regenerative Medicine, Section of Endocrinology, Diabetology and Metabolism, Di.Bi.M.I.S., University of Palermo, Piazza delle Cliniche 2, 90127, Palermo, Italy. carla.giordano@unipa.it. AD - ATeN (Advanced Technologies Network Center), University of Palermo, Palermo, Italy. carla.giordano@unipa.it. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20170703 PL - England TA - Stem Cell Res Ther JT - Stem cell research & therapy JID - 101527581 RN - 0 (Cytokines) SB - IM MH - Adult MH - Aged MH - CD8-Positive T-Lymphocytes/*immunology/pathology MH - Cells, Cultured MH - Cytokines/immunology MH - Female MH - Fibroblasts/*immunology/physiology MH - Hashimoto Disease/*immunology/pathology MH - Humans MH - *Immune Tolerance MH - Middle Aged MH - Stem Cells/*immunology/pathology MH - Th1 Cells/*immunology/pathology PMC - PMC5496215 OTO - NOTNLM OT - Hashimoto's thyroiditis OT - Human limbal stem cells OT - Immunoregulation OT - Inflammatory diseases OT - Tolerance induction EDAT- 2017/07/05 06:00 MHDA- 2018/03/27 06:00 PMCR- 2017/07/03 CRDT- 2017/07/05 06:00 PHST- 2017/04/07 00:00 [received] PHST- 2017/06/15 00:00 [accepted] PHST- 2017/06/02 00:00 [revised] PHST- 2017/07/05 06:00 [entrez] PHST- 2017/07/05 06:00 [pubmed] PHST- 2018/03/27 06:00 [medline] PHST- 2017/07/03 00:00 [pmc-release] AID - 10.1186/s13287-017-0611-5 [pii] AID - 611 [pii] AID - 10.1186/s13287-017-0611-5 [doi] PST - epublish SO - Stem Cell Res Ther. 2017 Jul 3;8(1):154. doi: 10.1186/s13287-017-0611-5.