Abstract
purpose. To study the role of TGF-β2 in posterior capsule opacification (PCO) and to determine whether CAT-152 (lerdelimumab), a fully human monoclonal antibody that neutralizes the effect of TGF-β2, can also provide therapeutic benefit for PCO.
methods. In vitro capsular bags were prepared from human donor eyes and maintained in a 5% CO2 atmosphere at 35°C. To investigate expression of active TGF-β2, capsular bags were incubated in serum-free EMEM for 2, 28, or more than 100 days and analyzed by ELISA (n ≥ 4 at each time point). To study underlying mechanisms, match-pair experiments were also performed, so that the medium was supplemented with 0, 1 or 10 ng/mL TGF-β2 with or without 10 μg/mL CAT-152 (n = 4 in all cases). On-going observations were by phase-contrast microscopy. In addition, donor material from patients who had undergone cataract surgery was analyzed. Cellular architecture was examined by fluorescence cytochemistry. Expression of matrix metalloproteinase (MMP)-2 and -9 was assessed by gelatin zymography.
results. Analysis of capsular bags from donor eyes that had received an intraocular lens (IOL) revealed the presence of endogenous active TGF-β2, matrix wrinkling, and expression of transdifferentiation markers αSMA and fibronectin. When cultured in vitro, donor bags also showed sustained release of MMP-2 and -9. Culture of capsular bags prepared in vitro from whole lenses showed that TGF-β2 (1–10 ng/mL) stimulated transdifferentiation and contraction of the capsular bag, resulting in light scatter. TGF-β2 also induced sustained release of MMP-2 and -9. Active TGF-β2 was detected in these cultures. The human monoclonal anti-TGF-β2 antibody CAT-152 (10 μg/mL) effectively inhibited all TGF-β2–induced effects.
conclusions. Addition of TGF-β2 accelerates transdifferentiation and contraction of the capsular bag, resulting in light scatter. CAT-152 inhibited all the effects of TGF-β2 that were examined and therefore has the potential to suppress development of PCO and provide potential therapeutic benefit to cataract patients.
methods. In vitro capsular bags were prepared from human donor eyes and maintained in a 5% CO2 atmosphere at 35°C. To investigate expression of active TGF-β2, capsular bags were incubated in serum-free EMEM for 2, 28, or more than 100 days and analyzed by ELISA (n ≥ 4 at each time point). To study underlying mechanisms, match-pair experiments were also performed, so that the medium was supplemented with 0, 1 or 10 ng/mL TGF-β2 with or without 10 μg/mL CAT-152 (n = 4 in all cases). On-going observations were by phase-contrast microscopy. In addition, donor material from patients who had undergone cataract surgery was analyzed. Cellular architecture was examined by fluorescence cytochemistry. Expression of matrix metalloproteinase (MMP)-2 and -9 was assessed by gelatin zymography.
results. Analysis of capsular bags from donor eyes that had received an intraocular lens (IOL) revealed the presence of endogenous active TGF-β2, matrix wrinkling, and expression of transdifferentiation markers αSMA and fibronectin. When cultured in vitro, donor bags also showed sustained release of MMP-2 and -9. Culture of capsular bags prepared in vitro from whole lenses showed that TGF-β2 (1–10 ng/mL) stimulated transdifferentiation and contraction of the capsular bag, resulting in light scatter. TGF-β2 also induced sustained release of MMP-2 and -9. Active TGF-β2 was detected in these cultures. The human monoclonal anti-TGF-β2 antibody CAT-152 (10 μg/mL) effectively inhibited all TGF-β2–induced effects.
conclusions. Addition of TGF-β2 accelerates transdifferentiation and contraction of the capsular bag, resulting in light scatter. CAT-152 inhibited all the effects of TGF-β2 that were examined and therefore has the potential to suppress development of PCO and provide potential therapeutic benefit to cataract patients.
| Original language | English |
|---|---|
| Pages (from-to) | 2301-2308 |
| Number of pages | 8 |
| Journal | Investigative Ophthalmology & Visual Science |
| Volume | 43 |
| Issue number | 7 |
| Publication status | Published - 2002 |