Document Detail

Induction of T regulatory cells by cytotoxic T-lymphocyte antigen-2α on corneal endothelial cells.
MedLine Citation:
PMID:  21245393     Owner:  NLM     Status:  MEDLINE    
PURPOSE: To determine whether murine corneal endothelial (CE) cells can promote the generation of T regulatory (Treg) cells in vitro.
METHODS: To induce Treg cells in vitro by CE cell lines, T cells exposed to CE cells were used as Treg cells. T cells exposed to CE cells in the presence of anti-mouse CD3 antibody were harvested and added to target bystander T cells in vitro. T-cell activation was assessed for proliferation by [(3)H]-thymidine incorporation. Expression of CD25 or Foxp3 on Treg cells was evaluated by flow cytometry. Expression of cytotoxic T-lymphocyte antigen-2 alpha (CTLA-2α) on CE cells was evaluated by flow cytometry, RT-PCR, immunohistochemistry, or in situ hybridization. Anti-CTLA-2α neutralizing antibodies, CTLA-2α siRNA, or pro-cathepsin L blocking proteins were used to abolish the CE-inhibitory function.
RESULTS: Cultured CE cells produced CTLA-2α on their surfaces, thereby enabling bystander CD4(+) T cells to be converted to Treg cells by TGFβ promotion. CE-induced Treg cells had immunosuppressive capacities by highly expressing CD25(high) and Foxp3. When mRNA downregulation (siRNA transfection), neutralizing antibodies, or blocking proteins were used to block CTLA-2α expression on CE cells, CE-induced Treg cells failed to acquire Treg function.
CONCLUSIONS: These findings indicate that cell surface CTLA-2α contributes to the CE-dependent suppression of bystander T cells. Thus, ocular resident tissue-exposed T cells can be induced to become regulators within the peripheral microenvironment.
Sunao Sugita; Yukiko Yamada; Shintaro Horie; Orie Nakamura; Kazumi Ishidoh; Yoshimi Yamamoto; Satoru Yamagami; Manabu Mochizuki
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2011-04-20
Journal Detail:
Title:  Investigative ophthalmology & visual science     Volume:  52     ISSN:  1552-5783     ISO Abbreviation:  Invest. Ophthalmol. Vis. Sci.     Publication Date:  2011 Apr 
Date Detail:
Created Date:  2011-04-21     Completed Date:  2011-06-29     Revised Date:  2013-01-09    
Medline Journal Info:
Nlm Unique ID:  7703701     Medline TA:  Invest Ophthalmol Vis Sci     Country:  United States    
Other Details:
Languages:  eng     Pagination:  2598-605     Citation Subset:  IM    
Department of Ophthalmology and Visual Science, Tokyo Medical and Dental University Graduate School of Medicine and Dental Sciences, 1-5-45 Yushima, Bunkyo-ku, Tokyo, Japan.
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MeSH Terms
Antibodies, Blocking / pharmacology
Antibodies, Neutralizing / pharmacology
Antigens, Differentiation / physiology*
Blotting, Western
Cathepsin L / antagonists & inhibitors
Cell Line, Transformed
Endothelium, Corneal / drug effects,  immunology*
Enzyme Precursors / antagonists & inhibitors
Flow Cytometry
Forkhead Transcription Factors / metabolism
In Situ Hybridization
Interleukin-2 Receptor alpha Subunit / metabolism
Lymphocyte Activation / physiology
Mice, Inbred C57BL
RNA, Small Interfering / pharmacology
Reverse Transcriptase Polymerase Chain Reaction
T-Lymphocytes / immunology
T-Lymphocytes, Regulatory / immunology*
Transforming Growth Factor beta / metabolism
Reg. No./Substance:
0/Antibodies, Blocking; 0/Antibodies, Neutralizing; 0/Antigens, Differentiation; 0/Enzyme Precursors; 0/Forkhead Transcription Factors; 0/Foxp3 protein, mouse; 0/Il2ra protein, mouse; 0/Interleukin-2 Receptor alpha Subunit; 0/RNA, Small Interfering; 0/Transforming Growth Factor beta; 0/cytotoxic T-lymphocyte antigen-2alpha, mouse; EC 3.4.22.-/procathepsin L; EC L

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