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October 1994, Vol 112, No. 10 >
ARTICLE | October 1994

Lysosomal Enzyme Activities in Conjunctival Tissues of Patients With Keratoconus FREE

Takeo Fukuchi, MD; Beatrice Y. J. T. Yue, PhD; Joel Sugar, MD; Sheridan Lam, MD
Arch Ophthalmol. 1994;112(10):1368-1374. doi:10.1001/archopht.1994.01090220118033
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Objective:  To examine the lysosomal enzyme activities in the conjunctival tissues of patients with keratoconus.

Methods:  Tissues collected from 11 patients with keratoconus, eight patients with senile cataract, three patients with Fuchs' corneal dystrophy, and 11 normal control subjects were processed for histochemical staining for two lysosomal hydrolases, acid esterase and acid phosphatase.

Results:  The epithelium of all conjunctival specimens stained positively for the two enzymes. The staining in the conjunctival tissues of patients with keratoconus was more prominent than that seen in specimens from either normal control subjects or patients with other diseases.

Conclusion:  Our results suggest that the conjunctival epithelium may be altered in keratoconus. Elevation of lysosomal enzyme levels has been demonstrated in the epithelium of corneas with keratoconus, implicating a role of this layer in the disease. The conjunctival abnormality seems to corroborate the corneal epithelial theory. It also adds one dimension to the pathogenesis of keratoconus.

REFERENCES

1 +
Krachmer JH, Feder RS, Belin MW.  Keratoconus and related non-inflammatory corneal disorders . Surv Ophthalmol . 1984;;28:293-322.
2 +
Bron AJ.  Keratoconus . Cornea . 1988;;7:163-169.
3 +
Macsai MS, Varley GA, Krachmer JH.  Development of keratoconus after contact lens wear: patient characteristics . Arch Ophthalmol . 1990;;108:534-538.
4 +
Sawaguchi S, Yue BYJT, Sugar J, Gilboy JE.  Lysosomal enzyme abnormalities in keratoconus . Arch Ophthalmol . 1989;;107:1507-1510.
5 +
Sawaguchi S, Twining SS, Yue BYJT, Wilson PM, Sugar J, Chan S-K.  A1-proteinase inhibitor levels in keratoconus . Exp Eye Res . 1990;;50:549-554.
6 +
Yue B, Loushin G, Sawaguchi S, Twining SS, Sugar J.  α2-Macroglobulin levels in keratoconus corneas . Invest Ophthalmol Vis Sci . 1992;;33( (suppl) ):976.
7 +
Fini ME, Yue BYJT, Sugar J.  Collagenolytic/gelatinolytic metalloproteinases in normal human and keratoconus corneas . Curr Eye Res . 1992;;9:849-862.
8 +
Brown D, Chwa MM, Opbroek A, Kenney MC.  Keratoconus corneas: increased gelatinolytic activity appears after modification of inhibitors . Curr Eye Res . 1993;;12:571-581.
9 +
Kao WWY, Vergnes JP, Ebert J, SundarRaj CV, Brown SI.  Increased collagenase and gelatinase activities in keratoconus . Biochem Biophys Res Commun . 1982;;107:929-936.
10 +
Rehany U, Lahav M, Shoshan S.  Collagenolytic activity in keratoconus . Ann Ophthalmol . 1982;;14:751-754.
11 +
Thoft RA, Wiley LA, SundarRaj N.  The multipotential cells of the limbus . Eye . 1989;;3:109-113.
12 +
Kitano S.  Ocular surface: physiological and pathological properties . Acta Soc Ophthalmol Jpn . 1987;;91:1-26.
13 +
Schermer A, Galvin S, Sun T-T.  Differentiation-related expression of a major 64K corneal keratin in vivo and in culture suggests limbal location of corneal epithelial stem cells . J Cell Biol . 1986;;103:49-62.
14 +
Cotsarelis G, Cheng S-Z, Dong G, Sun T-T, Lavker RM.  Existence of slowrecycling limbal epithelial basal cells that can be preferentially stimulated to proliferation: implications on epithelial stem cells . Cell . 1989;;57:201-209.
15 +
Tseng SCG.  Concept and application of limbal stem cells . Eye . 1989;;3:141-157.
16 +
Huang AJW, Tseng SCG.  Corneal epithelial wound healing in the absence of limbal epithelium . Invest Ophthalmol Vis Sci . 1991;;32:96-105.
17 +
Wiley L, SundarRaj N, Sun T-T, Thoft RA.  Regional heterogeneity in human corneal and limbal epithelia: an immunohistochemical evaluation . Invest Ophthalmol Vis Sci . 1991;;32:594-602.
18 +
Chung EH, Bukusoglu G, Zieske JD.  Localization of corneal epithelial stem cells in the developing rat . Invest Ophthalmol Vis Sci . 1992;;33:2199-2206.
19 +
Shapiro MS, Friend J, Thoft RA.  Corneal re-epithelization from the conjunctiva . Invest Ophthalmol Vis Sci . 1981;;21:135-142.
20 +
Kinoshita S, Friend J, Thoft RA.  Biphasic cell proliferation in transdifferentiation of conjunctival to corneal epithelium in rabbits . Invest Ophthalmol Vis Sci . 1983;;24:1008-1014.
21 +
Vanden Hoek TL, Goossens W, Knepper PA.  Fluorescence-labeled lectins, glycoconjugates, and the development of the mouse AOP . Invest Ophthalmol Vis Sci . 1987;;28:451-458.
22 +
Sando GN, Rosenbaum LM.  Human lysosomal acid lipase/cholesteryl ester hydrolase . J Biol Chem . 1985;;260:15185-15193.
23 +
Koster JF, Vaandrager H, Van Berkel TJC.  Study of the hydrolysis of 4-methylumbelliferyl oleate by acid lipase and cholesteryl oleate by acid cholesteryl esterase in human leucocytes, fibroblasts and liver . Biochim Biophys Acta . 1980;;618:98-105.
24 +
Sawaguchi S, Yue YJT, Kawa JE, Chang IL, Twining SS, Meberg B.  Lysosomal enzyme and inhibitor levels in the human trabecular meshwork . Invest Ophthalmol Vis Sci . 1994;;35:251-261.
25 +
Hayasaka S.  Lysosomal enzymes in ocular tissues and diseases . Surv Ophthalmol . 1983;;27:245-258.
26 +
Wei ZG, Wu RL, Lavker RM, Sun T-T.  In vitro growth and differentiation of rabbit bulbar, fornix and peripheral conjunctiva epithelia: implications on conjunctival epithelial transdifferentiation and stem cells . Invest Ophthalmol Vis Sci . 1993;;34:1814-1828.
27 +
Kolega J, Manabe M, Sun T-T.  Basement membrane heterogeneity and variation in corneal epithelial differentiation . Differentiation . 1989;;42:54-63.
28 +
Teng CC.  Electron microscopic study of the pathology of keratoconus, I . Am J Ophthalmol . 1963;;55:18-47.
29 +
Iwamoto T, DeVoe AG.  Particulate structures in keratoconus . Arch Ophthalmol . 1975;;35:65-76.
30 +
Zimmermann DR, Fisher RW, Winterhalter KH, Witmer R, Vaughan L.  Comparative studies of collagens in normal and keratoconus corneas . Exp Eye Res . 1988;;46:431-442.
31 +
Philipp W.  Altered lectin binding sites in keratoconus corneas . Curr Eye Res . 1992;;5:397-409.
32 +
Pouliquen Y.  Keratoconus . Eye . 1987;;1:1-14.
33 +
Patey A, Savoldelli M, Pouliquen Y.  Keratoconus and normal cornea: a comparative study of the collagenous fibers of the corneal stroma by image analysis . Cornea . 1984;;3:119-124.
34 +
Scroggs MW, Proia AD.  Histopathological variation in keratoconus . Cornea . 1992;;11:553-559.
35 +
Thoft RA.  The X, Y, Z hypothesis of corneal epithelial maintenance . Invest Ophthalmol Vis Sci . 1983;;24:1442-1443.
36 +
Alldredge OC, Krachmer JH.  Clinical types of corneal transplant rejection: their manifestations, frequency preoperative correlates, and treatment . Arch Ophthalmol . 1981;;99:599-604.
37 +
Abelson MB, Collin B, Gillette TE, Dohlman CH.  Recurrent keratoconus after keratoplasty . Am J Ophthalmol . 1980;;90:672-676.
38 +
Nirankari VS, Karesh J, Bastion F, Lakhanpal V, Billings E.  Recurrence of keratoconus in donor cornea 22 years after successful keratoplasty . Br J Ophthalmol . 1983;;67:23-28.
39 +
Wu WCS, Stark WJ, Green R.  Corneal wound healing after 193-nm excimer laser keratectomy . Arch Ophthalmol . 1991;;109:1426-1432.
40 +
Bechrakis N, Blom ML, Stark WJ, Green WR.  Recurrent keratoconus . Cornea . 1994;;12:73-77.
41 +
Johnson BL, Brown SI, Zeidman GW.  A light and electron microscopic study of recurrent granular dystrophy of the cornea . Am J Ophthalmol . 1981;;92:49-58.

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1 +
Krachmer JH, Feder RS, Belin MW.  Keratoconus and related non-inflammatory corneal disorders . Surv Ophthalmol . 1984;;28:293-322.
2 +
Bron AJ.  Keratoconus . Cornea . 1988;;7:163-169.
3 +
Macsai MS, Varley GA, Krachmer JH.  Development of keratoconus after contact lens wear: patient characteristics . Arch Ophthalmol . 1990;;108:534-538.
4 +
Sawaguchi S, Yue BYJT, Sugar J, Gilboy JE.  Lysosomal enzyme abnormalities in keratoconus . Arch Ophthalmol . 1989;;107:1507-1510.
5 +
Sawaguchi S, Twining SS, Yue BYJT, Wilson PM, Sugar J, Chan S-K.  A1-proteinase inhibitor levels in keratoconus . Exp Eye Res . 1990;;50:549-554.
6 +
Yue B, Loushin G, Sawaguchi S, Twining SS, Sugar J.  α2-Macroglobulin levels in keratoconus corneas . Invest Ophthalmol Vis Sci . 1992;;33( (suppl) ):976.
7 +
Fini ME, Yue BYJT, Sugar J.  Collagenolytic/gelatinolytic metalloproteinases in normal human and keratoconus corneas . Curr Eye Res . 1992;;9:849-862.
8 +
Brown D, Chwa MM, Opbroek A, Kenney MC.  Keratoconus corneas: increased gelatinolytic activity appears after modification of inhibitors . Curr Eye Res . 1993;;12:571-581.
9 +
Kao WWY, Vergnes JP, Ebert J, SundarRaj CV, Brown SI.  Increased collagenase and gelatinase activities in keratoconus . Biochem Biophys Res Commun . 1982;;107:929-936.
10 +
Rehany U, Lahav M, Shoshan S.  Collagenolytic activity in keratoconus . Ann Ophthalmol . 1982;;14:751-754.
11 +
Thoft RA, Wiley LA, SundarRaj N.  The multipotential cells of the limbus . Eye . 1989;;3:109-113.
12 +
Kitano S.  Ocular surface: physiological and pathological properties . Acta Soc Ophthalmol Jpn . 1987;;91:1-26.
13 +
Schermer A, Galvin S, Sun T-T.  Differentiation-related expression of a major 64K corneal keratin in vivo and in culture suggests limbal location of corneal epithelial stem cells . J Cell Biol . 1986;;103:49-62.
14 +
Cotsarelis G, Cheng S-Z, Dong G, Sun T-T, Lavker RM.  Existence of slowrecycling limbal epithelial basal cells that can be preferentially stimulated to proliferation: implications on epithelial stem cells . Cell . 1989;;57:201-209.
15 +
Tseng SCG.  Concept and application of limbal stem cells . Eye . 1989;;3:141-157.
16 +
Huang AJW, Tseng SCG.  Corneal epithelial wound healing in the absence of limbal epithelium . Invest Ophthalmol Vis Sci . 1991;;32:96-105.
17 +
Wiley L, SundarRaj N, Sun T-T, Thoft RA.  Regional heterogeneity in human corneal and limbal epithelia: an immunohistochemical evaluation . Invest Ophthalmol Vis Sci . 1991;;32:594-602.
18 +
Chung EH, Bukusoglu G, Zieske JD.  Localization of corneal epithelial stem cells in the developing rat . Invest Ophthalmol Vis Sci . 1992;;33:2199-2206.
19 +
Shapiro MS, Friend J, Thoft RA.  Corneal re-epithelization from the conjunctiva . Invest Ophthalmol Vis Sci . 1981;;21:135-142.
20 +
Kinoshita S, Friend J, Thoft RA.  Biphasic cell proliferation in transdifferentiation of conjunctival to corneal epithelium in rabbits . Invest Ophthalmol Vis Sci . 1983;;24:1008-1014.
21 +
Vanden Hoek TL, Goossens W, Knepper PA.  Fluorescence-labeled lectins, glycoconjugates, and the development of the mouse AOP . Invest Ophthalmol Vis Sci . 1987;;28:451-458.
22 +
Sando GN, Rosenbaum LM.  Human lysosomal acid lipase/cholesteryl ester hydrolase . J Biol Chem . 1985;;260:15185-15193.
23 +
Koster JF, Vaandrager H, Van Berkel TJC.  Study of the hydrolysis of 4-methylumbelliferyl oleate by acid lipase and cholesteryl oleate by acid cholesteryl esterase in human leucocytes, fibroblasts and liver . Biochim Biophys Acta . 1980;;618:98-105.
24 +
Sawaguchi S, Yue YJT, Kawa JE, Chang IL, Twining SS, Meberg B.  Lysosomal enzyme and inhibitor levels in the human trabecular meshwork . Invest Ophthalmol Vis Sci . 1994;;35:251-261.
25 +
Hayasaka S.  Lysosomal enzymes in ocular tissues and diseases . Surv Ophthalmol . 1983;;27:245-258.
26 +
Wei ZG, Wu RL, Lavker RM, Sun T-T.  In vitro growth and differentiation of rabbit bulbar, fornix and peripheral conjunctiva epithelia: implications on conjunctival epithelial transdifferentiation and stem cells . Invest Ophthalmol Vis Sci . 1993;;34:1814-1828.
27 +
Kolega J, Manabe M, Sun T-T.  Basement membrane heterogeneity and variation in corneal epithelial differentiation . Differentiation . 1989;;42:54-63.
28 +
Teng CC.  Electron microscopic study of the pathology of keratoconus, I . Am J Ophthalmol . 1963;;55:18-47.
29 +
Iwamoto T, DeVoe AG.  Particulate structures in keratoconus . Arch Ophthalmol . 1975;;35:65-76.
30 +
Zimmermann DR, Fisher RW, Winterhalter KH, Witmer R, Vaughan L.  Comparative studies of collagens in normal and keratoconus corneas . Exp Eye Res . 1988;;46:431-442.
31 +
Philipp W.  Altered lectin binding sites in keratoconus corneas . Curr Eye Res . 1992;;5:397-409.
32 +
Pouliquen Y.  Keratoconus . Eye . 1987;;1:1-14.
33 +
Patey A, Savoldelli M, Pouliquen Y.  Keratoconus and normal cornea: a comparative study of the collagenous fibers of the corneal stroma by image analysis . Cornea . 1984;;3:119-124.
34 +
Scroggs MW, Proia AD.  Histopathological variation in keratoconus . Cornea . 1992;;11:553-559.
35 +
Thoft RA.  The X, Y, Z hypothesis of corneal epithelial maintenance . Invest Ophthalmol Vis Sci . 1983;;24:1442-1443.
36 +
Alldredge OC, Krachmer JH.  Clinical types of corneal transplant rejection: their manifestations, frequency preoperative correlates, and treatment . Arch Ophthalmol . 1981;;99:599-604.
37 +
Abelson MB, Collin B, Gillette TE, Dohlman CH.  Recurrent keratoconus after keratoplasty . Am J Ophthalmol . 1980;;90:672-676.
38 +
Nirankari VS, Karesh J, Bastion F, Lakhanpal V, Billings E.  Recurrence of keratoconus in donor cornea 22 years after successful keratoplasty . Br J Ophthalmol . 1983;;67:23-28.
39 +
Wu WCS, Stark WJ, Green R.  Corneal wound healing after 193-nm excimer laser keratectomy . Arch Ophthalmol . 1991;;109:1426-1432.
40 +
Bechrakis N, Blom ML, Stark WJ, Green WR.  Recurrent keratoconus . Cornea . 1994;;12:73-77.
41 +
Johnson BL, Brown SI, Zeidman GW.  A light and electron microscopic study of recurrent granular dystrophy of the cornea . Am J Ophthalmol . 1981;;92:49-58.

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