Madhuri Amulya Koduri1,2, Jilu Jaffet1,2, Swapna S Shanbhag3, Sayan Basu4,5, Vivek Singh6, Swati Singh7. 1. Centre for Ocular Regeneration (CORE), Prof. Brien Holden Eye Research Centre (BHERC), LV Prasad Eye Institute, Road No.2, Banjara Hills, Hyderabad, 500034, Telangana, India. 2. Manipal Academy of Higher Education (MAHE), Karnataka, India. 3. The Cornea Institute, LV Prasad Eye Institute, Hyderabad, Telangana, India. 4. Centre for Ocular Regeneration (CORE), Prof. Brien Holden Eye Research Centre (BHERC), LV Prasad Eye Institute, Road No.2, Banjara Hills, Hyderabad, 500034, Telangana, India. sayanbasu@lvpei.org. 5. The Cornea Institute, LV Prasad Eye Institute, Hyderabad, Telangana, India. sayanbasu@lvpei.org. 6. Centre for Ocular Regeneration (CORE), Prof. Brien Holden Eye Research Centre (BHERC), LV Prasad Eye Institute, Road No.2, Banjara Hills, Hyderabad, 500034, Telangana, India. viveksingh@lvpei.org. 7. Centre for Ocular Regeneration (CORE), Prof. Brien Holden Eye Research Centre (BHERC), LV Prasad Eye Institute, Road No.2, Banjara Hills, Hyderabad, 500034, Telangana, India. swatisingh@lvpei.org.
Abstract
PURPOSE: To study the cytokeratin profile and keratinization-related gene expression in keratinized lid margins of chronic Stevens-Johnson syndrome (SJS) patients. METHODS: Posterior eyelid margins from 24 chronic SJS patients undergoing mucous membrane grafting and six healthy margins (orbital exenteration, fresh body donors) were studied using immunofluorescence staining (CK10, CK1, filaggrin, transglutaminase 1 (TGM1), (CK19, MUC5AC)) and quantitative PCR (keratinization-related genes-HBEGF, KGF, EGF, TGFα, TGFβ, and TNFα). The staining and gene expression were studied separately in the lid margin epidermis (LME) and lid margin conjunctiva (LMC). RESULTS: The expression of CK 1/10, filaggrin, and TGM1 in the LMC was similar to the LME in SJS patients. CK19 was expressed only in the basal epithelial layer of the LMC with loss of MUC5AC expression. Increased expression of KGF (p ≤ 0.056), TNFα (p ≤ 0.02), and TGFα (p ≤ 0.01) was observed in the LME of SJS patients compared to normal LME. LMC of SJS patients showed an increased expression of HBEGF (p ≤ 0.002), EGF (p ≤ 0.0002), KGF (p ≤ 0.02), TNFα (p ≤ 0.04), TGFα (p ≤ 0.003), and TGFβ (p ≤ 0.001) compared to normal LMC. Significant differences were observed in the expression of these genes between LME and LMC of SJS patients. These genes were validated using String analysis, which revealed the positive regulation of keratinization. CONCLUSION: In lid margins of SJS, there is an increased expression of keratinization-related genes compared to the normal lid margin. Keratinized LMC shares similar cytokeratin profile and keratinization gene expression as seen in cutaneous epithelium of SJS patients, indicating the possibility of the cutaneous epithelium as a source for keratinized LMC.
PURPOSE: To study the cytokeratin profile and keratinization-related gene expression in keratinized lid margins of chronic Stevens-Johnson syndrome (SJS) patients. METHODS: Posterior eyelid margins from 24 chronic SJS patients undergoing mucous membrane grafting and six healthy margins (orbital exenteration, fresh body donors) were studied using immunofluorescence staining (CK10, CK1, filaggrin, transglutaminase 1 (TGM1), (CK19, MUC5AC)) and quantitative PCR (keratinization-related genes-HBEGF, KGF, EGF, TGFα, TGFβ, and TNFα). The staining and gene expression were studied separately in the lid margin epidermis (LME) and lid margin conjunctiva (LMC). RESULTS: The expression of CK 1/10, filaggrin, and TGM1 in the LMC was similar to the LME in SJS patients. CK19 was expressed only in the basal epithelial layer of the LMC with loss of MUC5AC expression. Increased expression of KGF (p ≤ 0.056), TNFα (p ≤ 0.02), and TGFα (p ≤ 0.01) was observed in the LME of SJS patients compared to normal LME. LMC of SJS patients showed an increased expression of HBEGF (p ≤ 0.002), EGF (p ≤ 0.0002), KGF (p ≤ 0.02), TNFα (p ≤ 0.04), TGFα (p ≤ 0.003), and TGFβ (p ≤ 0.001) compared to normal LMC. Significant differences were observed in the expression of these genes between LME and LMC of SJS patients. These genes were validated using String analysis, which revealed the positive regulation of keratinization. CONCLUSION: In lid margins of SJS, there is an increased expression of keratinization-related genes compared to the normal lid margin. Keratinized LMC shares similar cytokeratin profile and keratinization gene expression as seen in cutaneous epithelium of SJS patients, indicating the possibility of the cutaneous epithelium as a source for keratinized LMC.
Authors: Erich Knop; Nadja Knop; Andrey Zhivov; Robert Kraak; Donald R Korb; Caroline Blackie; Jack V Greiner; Rudolf Guthoff Journal: J Anat Date: 2011-04 Impact factor: 2.610