PMID- 21498621
OWN - NLM
STAT- MEDLINE
DCOM- 20110928
LR  - 20220410
IS  - 1552-5783 (Electronic)
IS  - 0146-0404 (Linking)
VI  - 52
IP  - 8
DP  - 2011 Jul 25
TI  - Hyperosmolar stress upregulates HLA-DR expression in human conjunctival 
      epithelium in dry eye patients and in vitro models.
PG  - 5488-96
LID - 10.1167/iovs.11-7215 [doi]
AB  - PURPOSE: To investigate the immune response of human conjunctival epithelium to 
      hyperosmolar stress. METHODS: Tear osmolarity was measured in 15 normal subjects 
      and 25 dry eye (DE) patients; conjunctival imprint cytology samples were obtained 
      at the nasal bulbar area. Subconfluent primary human conjunctival epithelial 
      cells (pHCECs) and human conjunctival organ cultures (hCOCs), both cultured in 
      iso-osmolar medium (305 mOsm/L), were exposed for 24 hours to media with 
      progressively higher osmolarity, with or without the ion channel inhibitor 
      ruthenium red (RuR). Human leukocyte antigen (HLA)-DR expression was evaluated by 
      immunocytochemistry, on imprints from subjects, on primary human conjunctival 
      epithelial cells, on formalin fixed-paraffin embedded hCOCs, and by RT-PCR. 
      Statistical evaluation was performed by applying the unpaired Student's t test, 
      as well as Spearman's rho and Pearson's r correlation coefficients (significance 
      P < 0.05). RESULTS: HLA-DR expression increased in DE subjects with respect to 
      control (% mean +/- SD, respectively, 46.16 +/- 7.2 vs. 7.48 +/- 1.14, P < 0.0001) and 
      exhibited significantly high correlations with tear osmolarity values (r = 0.614; 
      P < 0.0001). In vitro experiments showed a progressive increase in HLA-DR 
      expression as the osmolarity of the medium was increased from 6.75 +/- 1.16 (% mean 
      +/- SD) in iso-osmolar-cultured cells to 9.96 +/- 1.37 and 12.94 +/- 4.04 in cells 
      cultured in, respectively, 350 and 400 mOsm/L (P < 0.05). A stepwise progressive 
      increase was also found in hCOCs. Results were confirmed by RT-PCR. Ruthenium red 
      significantly reduced HLA-DR expression in hyperosmolar-cultured cells. 
      CONCLUSIONS: Data from complementary techniques demonstrate that extracellular 
      hyperosmolarity induces HLA-DR overexpression in human conjunctival epithelial 
      cells in both DE patients and in vitro cell culture models.
FAU - Versura, Piera
AU  - Versura P
AD  - Ophthalmology Unit, University of Bologna, Bologna, Italy. piera.versura@unibo.it
FAU - Profazio, Vincenzo
AU  - Profazio V
FAU - Schiavi, Costantino
AU  - Schiavi C
FAU - Campos, Emilio C
AU  - Campos EC
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20110725
PL  - United States
TA  - Invest Ophthalmol Vis Sci
JT  - Investigative ophthalmology & visual science
JID - 7703701
RN  - 0 (HLA-DR Antigens)
RN  - 0 (RNA, Messenger)
RN  - 0 (Saline Solution, Hypertonic)
RN  - 11103-72-3 (Ruthenium Red)
SB  - IM
MH  - Adult
MH  - Cells, Cultured
MH  - Conjunctiva/*metabolism
MH  - Dry Eye Syndromes/*genetics/metabolism
MH  - Epithelial Cells/drug effects/metabolism
MH  - Female
MH  - Gene Expression Regulation/*drug effects
MH  - HLA-DR Antigens/*genetics/*metabolism
MH  - Humans
MH  - Immunohistochemistry
MH  - Male
MH  - Middle Aged
MH  - Organ Culture Techniques
MH  - Osmolar Concentration
MH  - RNA, Messenger/metabolism
MH  - Reverse Transcriptase Polymerase Chain Reaction
MH  - Ruthenium Red/chemistry
MH  - Saline Solution, Hypertonic/*pharmacology
MH  - Tears/*metabolism
EDAT- 2011/04/19 06:00
MHDA- 2011/09/29 06:00
CRDT- 2011/04/19 06:00
PHST- 2011/04/19 06:00 [entrez]
PHST- 2011/04/19 06:00 [pubmed]
PHST- 2011/09/29 06:00 [medline]
AID - iovs.11-7215 [pii]
AID - 10.1167/iovs.11-7215 [doi]
PST - epublish
SO  - Invest Ophthalmol Vis Sci. 2011 Jul 25;52(8):5488-96. doi: 10.1167/iovs.11-7215.