November 1993
Volume 34, Issue 12
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Articles  |   November 1993
The antiproliferative effect of a transferrin-toxin on human retinal pigment epithelial cells and rabbit fibroblasts.
Author Affiliations
  • J T Handa
    Duke Eye Center, Durham, North Carolina.
  • L L Houston
    Duke Eye Center, Durham, North Carolina.
  • G J Jaffe
    Duke Eye Center, Durham, North Carolina.
Investigative Ophthalmology & Visual Science November 1993, Vol.34, 3419-3428. doi:
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      J T Handa, L L Houston, G J Jaffe; The antiproliferative effect of a transferrin-toxin on human retinal pigment epithelial cells and rabbit fibroblasts.. Invest. Ophthalmol. Vis. Sci. 1993;34(12):3419-3428.

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Abstract

PURPOSE: To determine the effect of a rabbit transferrin conjugated to recombinant ricin A chain (Tfr-rRA) and the carboxylic ionophore monensin on proliferating and density-arrested human retinal pigment epithelial cells and rabbit dermal fibroblasts. METHODS: Cells were seeded on 24-well plates at 20,000 cells/cm2 and exposed to Tfr-rRA (0.1-10,000 ng/ml) with or without monensin (0.01 microM), and with or without human transferrin (65.7 mg/l) for 5 minutes to 7 days. Cells were studied morphologically and counted at 1, 2, 4, and 7 days. RESULTS: Tfr-rRA (10-10,000 ng/ml) killed proliferating human retinal pigment epithelial cells and rabbit dermal fibroblasts in a dose-dependent manner (p < or = 0.01) up to a maximum of 86% and 93%, respectively. In contrast, Tfr-rRA had minimal effect on density-arrested human retinal pigment epithelial cells and rabbit dermal fibroblasts. The cytotoxicity of Tfr-rRA was inhibited by the addition of human transferrin (65.7 mg/l), an effect that was partially overcome by longer treatment with Tfr-rRA. Monensin (0.01 microM) increased the cytotoxicity of Tfr-rRA by 4.8-fold over Tfr-rRA alone, shortened the onset of cell kill with Tfr-rRA from 48 to 24 hours (P = 0.04), and partially reversed the neutralizing effect of human transferrin. CONCLUSIONS: The results indicate that Tfr-rRA effectively inhibited the proliferation of human retinal pigment epithelial cells and rabbit dermal fibroblasts in vitro. The inhibitory effect could be modified by the addition of human transferrin or monensin. Thus, this ricin A chain conjugate may interrupt the proliferation of cells necessary in the pathogenesis of proliferative vitreoretinopathy.

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