Objective:To characterize chemokine receptor CCR5 expression on the conjunctival epithelium in dry eye syndromes.Methods:Conjunctival impression cytology samples were obtained from normal subjects(n=15)and patients wi...Objective:To characterize chemokine receptor CCR5 expression on the conjunctival epithelium in dry eye syndromes.Methods:Conjunctival impression cytology samples were obtained from normal subjects(n=15)and patients with dry eye syndrome(n=45).Cells were harvested from impression cytology samples,and flow cytometry was performed to quantitatively analyze the cell surface expression of chemokine receptor CCR5.Characterization of CCR5-positive cells was done by 2-color flow cytometry using fluorescein-conjugated anti-CCR5 and phycoerythrin-conjugated anti-CD45 antibodies(whereCD45 is a marker for bone marrow-derivedcells).To study CCR5 messenger RNA transcripts,real-time polymerase chain reaction was done on RNA isolated from the impression cytology samples of normal subjects(n=5)and patients with dry eye syndrome(n=14).Results:We found significant upregulation in cell surface expression of CCR5 in patients with both aqueous tear-deficient and evaporative forms of dry eye syndrome(P <.001).The real-time polymerase chain reaction results(for messenger RNA)corroborated the flow cytometry data(for protein).The majority of the cells expressing CCR5 were non-bone marrow-derived resident epithelial cells of the conjunctiva.Conclusion:Our findings suggest that CCR5 up-regulation is significantly associated with dry eye syndrome-associated ocular surface disease.Clinical Relevance:Chemokine receptor CCR5 or its ligands may serve as useful targets for modulation of tissue immunoinflammatory responses in dry eye syndromes.展开更多
文摘Objective:To characterize chemokine receptor CCR5 expression on the conjunctival epithelium in dry eye syndromes.Methods:Conjunctival impression cytology samples were obtained from normal subjects(n=15)and patients with dry eye syndrome(n=45).Cells were harvested from impression cytology samples,and flow cytometry was performed to quantitatively analyze the cell surface expression of chemokine receptor CCR5.Characterization of CCR5-positive cells was done by 2-color flow cytometry using fluorescein-conjugated anti-CCR5 and phycoerythrin-conjugated anti-CD45 antibodies(whereCD45 is a marker for bone marrow-derivedcells).To study CCR5 messenger RNA transcripts,real-time polymerase chain reaction was done on RNA isolated from the impression cytology samples of normal subjects(n=5)and patients with dry eye syndrome(n=14).Results:We found significant upregulation in cell surface expression of CCR5 in patients with both aqueous tear-deficient and evaporative forms of dry eye syndrome(P <.001).The real-time polymerase chain reaction results(for messenger RNA)corroborated the flow cytometry data(for protein).The majority of the cells expressing CCR5 were non-bone marrow-derived resident epithelial cells of the conjunctiva.Conclusion:Our findings suggest that CCR5 up-regulation is significantly associated with dry eye syndrome-associated ocular surface disease.Clinical Relevance:Chemokine receptor CCR5 or its ligands may serve as useful targets for modulation of tissue immunoinflammatory responses in dry eye syndromes.
