Quantification of tear proteins and sPLA2-IIa alteration in patients with allergic conjunctivitis.
OBJECTIVE
Allergic conjunctivitis (AC) has been reported to induce the instability of the tear film. The tear protein and the lipid layer play important roles in maintaining the tear film. The aim of this study was to quantify the alteration of the major tear protein components and a lipid related protein secretory type IIa phospholipase A2 (sPLA2-IIa) in tears of seasonal allergic conjunctivitis (SAC) and perennial allergic conjunctivitis (PAC) patients.


METHODS
Twenty-one SAC and PAC patients and thirteen normal controls completed a symptom questionnaire and underwent regular ocular examination. SAC and PAC patients were diagnosed based on the clinical presentation and elevated serum IgE levels. Schirmer test paper was used to collect tear samples from SAC and PAC patients and normal controls. Soybean trypsin inhibitor (SBTI) was used as an internal standard to analyze tear samples in 15% SDS-PAGE gel. Total tear protein and its major components from the SAC and PAC patients and normal controls were quantified by band densitometry. The major tear protein bands were determined by MALDI-TOF/TOF spectrum analysis. Western blot was used to detect the content of sPLA2-IIa in tears of allergic conjunctivitis patients and normal controls.


RESULTS
Schirmer test scores were more than 10 mm in all the SAC and PAC patients and control subjects. The tear film breakup time of SAC and PAC patients was much shorter than that of the normal controls. We obtained 15 bands of tear protein by one dimensional SDS-PAGE, in which 14 bands were determined by mass-spectrum analysis. The band densitometry analysis revealed that the total tear protein concentration was much higher in SAC and PAC patients than in normal controls (p<0.05). The quantity of tear protein band 4 (serum albumin precursor), band 6 (Ig gamma-2), band 9 (leukocyte elastase inhibitor) were also significantly higher in AC patients (p<0.05). Content of sPLA2-IIa, as shown by western blot, was much higher in AC patients than in controls.


CONCLUSIONS
The total tear protein concentration and some of the major tear protein components was increased in tears of SAC and PAC patients. In addition, the content of sPLA2-IIa in tears of SAC and PAC patients was elevated. The tear protein changes in SAC and PAC patients may contribute to instability of tear film.
Zhongshan Ophthalmic Center, State Key Laboratory of Ophthalmology, Sun Yat-sen University, Guangzhou, China.
Kaijun
K
Li
Xialin
X
Liu
Ziyan
Z
Chen
Qiang
Q
Huang
Kaili
K
Wu
D000293:Adolescent; D000328:Adult; D016022:Case-Control Studies; D003233:Conjunctivitis, Allergic / Q000201:enzymology* / Q000473:pathology; D004591:Electrophoresis, Polyacrylamide Gel; D005123:Eye / Q000201:enzymology / Q000473:pathology; D005136:Eye Proteins / Q000737:chemistry / Q000378:metabolism*; D005260:Female; D054501:Group II Phospholipases A2 / Q000378:metabolism*; D006801:Humans; D008297:Male; D012015:Reference Standards; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D011795:Surveys and Questionnaires; D013666:Tears / Q000201:enzymology*; D055815:Young Adult
D005136:Eye Proteins; C042434:tear proteins; D054501:Group II Phospholipases A2; C516287:PLA2G2A protein, human