Quantification of Soluble Cytokines in Sjögren Disease, non Sjögren dry eye

and Healthy Subjects

M. Jaimes1B, C. Santacruz-Valdes1B, Y. Garfias1A, T. Ramirez-Luquin1B, M. Jimenez-Martinez2.

1Institute of Ophthalmology, “Fundacion Conde de Valenciana”, Mexico City, Mexico; AResearch Unit - Department of Immunology, B Department of Cornea and External Diseases; 2Biochemistry Department, Facultad de Medicina, Universidad Nacional Autonoma de Mexico, Mexico City, Mexico.

The authors have no financial interest in the subject matter of this poster.

Introduction

• The tear film is a complex fluid composed of a lot of proteins, cytokines among others.

• One of the most severe inflammatory eye diseases is Sjögren syndrome. This is an autoimmune disease that affects ocular surface

• It has been previously identified by ELISA and PCR that in Sjögren syndrome exists an up-regulation of cytokines and in this report we use cytometric bead arrays (CBA) cytokine detection kit to identifiy tear film cytokines in Sjögren syndrome, non Sjögren dry eye and healthy subjects.

Purpose

• To identify soluble cytokine concentrations in tear samples of patients with Sjögren disease, non Sjögren dry eye and healthy subjects.

*J Cataract Refract Surg 2009; 35:1101–1120

Materials and methods

• Patients classified according to tear film break-up time (BUT) <7 sec, Schirmer II test (ST) <7 mm, symptoms <12 points in a validated questionnaire and oral mucosa biopsy or SS-A, SS-B Ab in the Sjögren disease patients.

• We obtained a tear sample of both eyes and performed quantification of protein concentration. Using CBA Cytokine Detection Kit we performed IL-2, TNF-, IL-10, IL-4, IL-5 and IFN- detection in 50 l of tear sample.

• Descriptive statistics for demographic variables and ANOVA for identification of differences between groups. p was statistically significant if ≤0.05.

Results

• 65 patients were included, mean age 44 years (18-83), 37% male (n=24) and 63% female (n=41). Sjögren disease group showed the greatest concentration of TNF- and IFN- concentration (p < 0.05).

• We did not find statistically significant difference between non Sjögren dry eye and healthy subjects.

Results

Group Age Symptoms BUT Schirmer II

Healthy 33.3 (22,58) 6.36±4.22 9.66±1.54 18.36±8.74

Non Sjögren dry eye 44.3(18-83) 16.29±10.13 5.38±3.03 12.92±7.89

Sjögren 55.36 (41,85) 29.45±14.98* 2.84±1.03* 1.95±1.19*

*p<0.05

Results

IFN- TNF- IL-10 IL-5 IL-4 IL-2

Sjögren 37.94* 9.97* 7.43* 8.75 8.73* 5

SD 64.87 14.60 12.34 19.41 15.01 9.42

Non Sjögren dry eye 7.64 2 2.59 1.28 1.37 1.50

SD 29.39 5.56 7.69 3.74 6.01 5.23

Healthy 6.31 1.65 2.14 1.06 1.13 1.24

SD 26.75 5.09 7.03 3.42 5.46 4.77

*p<0.05

Results

Results

Results

Conclusions

• Sjögren patients showed higher cytokine concentration of TNF- and IFN- in the tear film.

• There was a statistically significative difference in IL-4 in Sjögren patients and healthy subjects only.

• Interesting, Sjögren patients showed the clinically most severe dry eye, this finding was not observed in the non Sjögren dry eye.

• Corresponding author: Martha Jaimes Gutiérrez• martha.jaimes@institutodeoftalmologia.org

Bibliography

• Ahn DH, Crawley SC, Hokari R, Yang SC, Li JD, Kim YS. TNF-alpha Activates MUC2 Transcription NF-kappaB but Inhibits via JNK Activation. Cell Physiol Biochem, 2005; 15:29-40.

• Pflugfelder SC, Jones D, Ji Z, Alfonso A, Monroy D. Altered Cytokine Balance in the Tear Fluid and Conjunctiva of Patients with Sjögren’s Syndrome Keratoconjunctivitis Sicca. Curr Eye Res, 1999: 19(3):201-211.

• Lemp MA. Report of the National Eye Institute/Industry Workshop on Clinical Trials in Dry Eyes. CLAO J, 1995; 21:221-232.

• Johnson ME, Murhpy PJ. Changes in the Tear Film and Ocular Surface From Dry Eye Syndrome. Prog Ret Eye Res 2004; 23: 449-74.