The antigenicity of ex vivo cultivated human corneal limbal epithelial and stromal cells: temporal changes in vitro.
OBJECTIVE
To investigate the effect of ex vivo culture on the expression of human leukocyte antigen (HLA) in human corneal limbal epithelial cells (LECs), and to evaluate the expression of HLA and costimulatory molecules on human corneal limbal stromal cells (LSCs).


METHODS
The expression of HLA-ABC and HLA-DR were evaluated using flow cytometry on cultured human LECs and LSCs after serial passage and compared with that on freshly isolated cells. Additionally, the expression of costimulatory molecules CD80, CD86, CD40, and immunoregulatory molecule HLA-G were investigated on LSCs with or without an addition of interferon (IFN) γ (250 U/mL, 4 days).


RESULTS
HLA-ABC and HLA-DR were expressed in 99.12 ± 1.02% and 25.86 ± 4.34% of freshly isolated LECs, respectively. After culture, the HLA-DR-expressing cells significantly decreased with serial passage, whereas the percentage of HLA-ABC-expressing cells did not change. More than 90% of LSCs expressed HLA-ABC and HLA-G, whereas the expression of HLA-DR, CD80, CD86, and CD40 was negligible. This expression profile did not change after serial passage. However, the treatment with IFN-γ induced a significant increase in HLA-DR (0.17% vs. 17.68%) and CD40 (0.19% vs. 13.19%) expression on LSCs.


CONCLUSIONS
The immunogenicity of LECs can be lowered after ex vivo cultivation, whereas LSCs can be immunogenic in the presence of IFN-γ.
Oh
Joo Youn
JY
Department of Ophthalmology, Seoul National University College of Medicine, Seoul, Korea.
Ko
Jung Hwa
JH
Lee
Hyun Ju
HJ
Kim
Mee Kum
MK
Lee
Jin Hak
JH
Wee
Won Ryang
WR
D015703:Antigens, CD / Q000032:analysis; D002478:Cells, Cultured; D003319:Corneal Stroma / Q000166:cytology / Q000276:immunology*; D005434:Flow Cytometry; D006680:HLA Antigens / Q000032:analysis* / Q000187:drug effects; D006684:HLA-DR Antigens / Q000378:metabolism; D006801:Humans; D007371:Interferon-gamma / Q000494:pharmacology; D016850:Limbus Corneae / Q000166:cytology / Q000276:immunology*; D013997:Time Factors
D015703:Antigens, CD; D006680:HLA Antigens; D006684:HLA-DR Antigens; D007371:Interferon-gamma