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. 2001 Nov;85(11):1362–1366. doi: 10.1136/bjo.85.11.1362

Latent TGFβ binding protein-1 and fibrillin-1 in human capsular opacification and in cultured lens epithelial cells

S Saika 1, T Miyamoto 1, T Tanaka 1, I Ishida 1, Y Ohnishi 1, A Ooshima 1
PMCID: PMC1723781  PMID: 11673307

Abstract

BACKGROUND/AIM—It was previously reported that collagenous extracellular matrix (ECM) in human capsular opacification contained isoforms of transforming growth factor β (TGFβ). In the present study, the authors performed immunohistochemistry to examine whether ECM in human capsular opacification and in cultures of bovine lens epithelial cells (LECs) contained latent TGFβ binding protein-1 (LTBP-1), TGFβ1 latency associated peptide (β1-LAP), and fibrillin-1, a suspected ligand of LTBP-1 as well as a component of the extracellular microfibrillar apparatus. The aim of the study was to further clarify the mechanism of TGFβ1 deposition in ECM of capsular opacification.
METHODS—Human capsular opacification specimens and uninjured lens capsules, as well as cultured bovine LECs, were processed for immunohistochemistry using antibodies against LTBP-1, β1-LAP, fibrillin-1, and collagen type I.
RESULTS—LTBP-1, β1-LAP, and fibrillin-1 all were localised to the ECM in human capsular opacification. Uninjured lens epithelium stained for β1-LAP, but not for LTBP-1 and fibrillin-1. ECM deposited in confluent LEC cultures stained for LTBP-1, β1-LAP, and fibrillin-1, while cultures with only sparse cellularity were unstained for LTBP-1 or fibrillin-1.
CONCLUSIONS—LECs upregulate LTBP-1 and fibrillin-1 during postoperative healing. LTBP-1, β1-LAP, and fibrillin-1 colocalised to the ECM in capsular opacification and in confluent LEC cultures. TGFβ1 is considered to deposit in ECM in the large latent form. ECM secreted by LEC may function as a scavenger or repository of TGFβ.



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Figure 1  .

Figure 1  

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Localisation pattern of large transforming growth factor β binding protein-1 (LTBP-1) in human posterior capsule opacification. In (A) (case 1) and (B) (case 8), lens epithelium of an uninjured lens capsule (A, arrowheads) and a capsule 10 days after cataract surgery (B, arrowheads) lack LTBP-1. In (C) (case 17), extracellular matrix accumulation (asterisk) between the anterior lens capsule and the anterior surface of the intraocular lens shows immunoreactivity for LTBP-1. Lens cells are negative for this protein (arrowheads). In (D) (case 19), lens cells (arrowheads) and regenerated lenticular fibres of Sommerring's ring unstained for LTBP-1. AC = anterior capsule; IOL = intraocular lens. Indirect immunostaining; bar = 20 µm.

Figure 2  .

Figure 2  

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Localisation pattern of β1 latency associated peptide (β1-LAP) in human posterior capsule opacification. In (A) (case 1), lens epithelial cells (arrowheads) of an uninjured lens capsule exhibit immunoreactivity for β1-LAP. In (B) (case 25), extracellular matrix accumulation (asterisk) around the edge of the capsulotomy (yellow arrow) shows weak immunoreactivity for β1-LAP, while lens cells (arrowheads) exhibit marked immunoreactivity. In (C) (case 17), extracellular matrix accumulation (asterisk) between the intraocular lens optic and the anterior capsule as well as lens cells (arrowheads) show strong β1-LAP immunoreactivity. AC = anterior capsule; IOL = intraocular lens. Indirect immunostaining; bar = 20 µm.

Figure 3  .

Figure 3  

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Fibrillin-1 as a component of fibrous extracellular matrix in human posterior capsule opacification. In (A) (case 2), uninjured lens epithelium (arrowheads) lacks immunoreactivity for fibrillin-1. In (B) (case 18), extracellular matrix accumulation (asterisk) around the edge of anterior capsulotomy (yellow arrow) shows marked immunoreactivity for fibrillin-1. In (C) (case 17), weak immunoreactivity for fibrillin-1 (arrows) is detected surrounding proliferating lens epithelial cells, which lack immunoreactivity. AC = anterior capsule; IOL = intraocular lens. Indirect immunostaining; bar = 20 µm.

Figure 4  .

Figure 4  

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Immunolocalisation of LTBP-1, β1-LAP, and fibrillin-1 in the extracellular matrix accumulation in bovine lens epithelial cell cultures. In sparse, preconfluent, cultures at day 3, almost no immunoreactivity for LTBP-1 was detected in LECs (A). In early confluence, a small amount of LTBP-1 was detected extracellularly in a very fine fibrillar pattern (B), while in mature confluent cultures at day 14, immunoreactivity for LTBP-1 appeared to form an organised network throughout the culture (C). Immunoreactivity for β1-LAP was detected in the cells of a sparsely cellular culture (D) and an early confluent culture (E). The mature confluent culture of LECs exhibited extracellular deposition of β1-LAP (F). In sparsely cellular cultures, LECs did not express fibrillin-1 protein (H). In early confluent cultures at day 10, fibrillin-1 was detected in a fine fibrillar pattern in fibroblast ECM; the cells themselves were unstained (I). Mature confluent LEC cultures showed a condensed fibrillin-1 structure (J). No specific immunoreactivity was observed in negative controls with anti-goat IgG (G) or anti-rabbit IgG but no primary antibody (K). LTBP-1 = large transforming growth factor β binding protein-1; β1-LAP = β1-latency associated peptide; LEC = lens epithelial cells. Indirect immunostaining; bar = 50 µm.

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