Ultrastructural and immunohistological characterization of the sirc corneal cell line

Niederkorn, Jérry Y.; Meyer, Dale R.; Ubelaker, John E.; Martin, J. Holmes

doi:10.1007/bf02624618

Cited by 55 publications

(32 citation statements)

References 17 publications

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“…Although in experimental models, SIRCs are described and used as being of epithelial origin [26][27][28], their morphology, ultrastructure and antigenic properties indicate a fibroblast origin for this cell line [29].…”

Section: Introductionmentioning

confidence: 99%

Antioxidant protection in cultured corneal cells and whole corneas submitted to UV-B exposure

Ciuffi

Pisanello

Pagliai

et al. 2003

Journal of Photochemistry and Photobiology B: Biology

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Several corneal pathologies are characterized by the presence of reactive oxygen species (ROS); therefore, we evaluated the protection afforded by pirenoxine and melatonin to corneal cell culture and whole rabbit cornea from ultraviolet exposure and other oxidant systems.Rabbit cornea cell (SIRC) plates and whole corneas were exposed to UV-B (80 or 800 mJ/cm 2 ) or incubated with fMLPstimulated autologous macrophages, in the presence or absence of pirenoxine or melatonin (10 À5 M). The protective activity of compounds was assessed by measuring superoxide anion formation, inhibition of oxidation and mitochondrial viability. Moreover the ex vivo protective effect of pirenoxine and melatonin was verified in the whole cornea submitted to UV-B exposure in vitro.Our experimental data demonstrate that pirenoxine and melatonin were able to inhibit the superoxide formation and oxidative effect in cell culture and whole rabbit corneas submitted to UV-B exposure or to incubation with fMLP-stimulated autologous macrophages.Mitochondrial viability was restored in epithelial cells of rabbit cornea but not in SIRCs. Moreover, both compounds are also able to increase ex vivo epithelial corneal cell defences against the in vitro UV-B induced lipid peroxidation.

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Section: Introductionmentioning

confidence: 99%

Antioxidant protection in cultured corneal cells and whole corneas submitted to UV-B exposure

Ciuffi

Pisanello

Pagliai

et al. 2003

Journal of Photochemistry and Photobiology B: Biology

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“…An objective of this research was to investigate whether the Statens Seruminstitut rabbit corneal (SIRC) cell line can serve as an effective in vitro screening tool in identifying nucleoside drugs with affinity for the nucleoside transporter expressed on the cornea. The SIRC cell line was originally derived from the cornea of a normal hare 24 and has been widely used as an in vitro model to assess corneal transport as well as immunological and toxicological responses. [25][26][27][28][29][30][31] However, contradictory reports exist about the epithelial origin of the cell line.…”

Section: Introductionmentioning

confidence: 99%

Functional differences in nucleoside and nucleobase transporters expressed on the rabbit corneal epithelial cell line (SIRC) and isolated rabbit cornea

et al. 2003

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ABSTRACTclovir permeates the cornea primarily by passive diffusion. Nucleoside and nucleobase transporters on rabbit cornea and corneal epithelial cell line, SIRC, are functionally different, undermining the utility of the SIRC cell line as an in vitro screening tool for elucidating the corneal permeation mechanism of nucleoside analogs. Uptake and transport studies were performed for 2 minutes and 120 minutes, respectively, at 34°C. Thymidine uptake by SIRC displayed saturable kinetics (K m = 595.9 ± 80.4μM, and V max = 289.5 ± 17.2 pmol/min/mg protein). Uptake was inhibited by both purine and pyrimidine nucleosides but not by nucleobases. [ 3 H]thymidine uptake was sodium and energy independent but was inhibited by nitrobenzylthioinosine at nanomolar concentrations. Adenine uptake by SIRC consisted of a saturable component (K m = 14.4 ± 2.3μM, V max = 0.4 ± 0.04 nmol/min/mg protein) and a nonsaturable component. Uptake of adenine was inhibited by purine nucleobases but not by the nucleosides or pyrimidine nucleobases and was independent of sodium, energy, and nitrobenzylthioinosine. [ 3 H]ganciclovir uptake involved a carrier-mediated component and was inhibited by the purine nucleobases but not by the nucleosides or pyrimidine nucleobases. However, transport of [ 3 H]adenine across the isolated rabbit cornea was not inhibited by unlabeled adenine. Further, corneal permeability of ganciclovir across a 100-fold concentration range remained constant, indicating that ganci-

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“…reported here were not described. SIRC is of rabbit corneal epithelium origin, but is biochemically more fibroblastic than it is epithelial (Niederkorn et al 1990), while MDCK cells polarize and differentiate in culture. We have tested several cell lines for their ability to produce highly organized multicellular structures when infected with V. corneae.…”

Section: Discussionmentioning

confidence: 99%

Multinucleate host cells induced by Vittaforma corneae (Microsporidia)

Leitch¹,

Shaw²,

Colden-Stanfield³

et al. 2005

FOLIA PARASIT

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Abstract. The microsporidium Vittaforma corneae (Shadduck, Meccoli, Davis et Font, 1990) develops within the target cell cytoplasm. In the present study, green monkey kidney (E6) cells infected at 30°C, 35°C or 37°C with V. corneae developed enlarged multinucleate structures of up to 200 µm in any horizontal dimension made up either of a single cell or of multiple fused cells. A number of epithelial cell types (SW-480, HT-29, Caco-2 and HCT-8) were infected with V. corneae but did not induce the same highly organized structures, suggesting that for the structure to develop, the host cell must be capable of continued mitosis, and not be differentiated or be detaching from the surface matrix. Live cell imaging of infected E6 cells revealed large, multinucleate infected cells characterized by a central focus from which radiated parasite stages and host cell mitochondria. Immunocytochemistry identifying γ and α tubulin suggested that a single centrally-located microtubule organizing centre governed the distribution of parasite stages and host cell organelles, with mitochondria and parasites being eventually transported towards the periphery of the structure. Whole cell patch clamp analysis of infected cells indicated an average five-fold increase in total membrane capacitance, consistent with an enlarged single cell. Scanning electron microscopy revealed cell-like protrusions around the periphery of the structure with the intervening space being made up of parasites and cell debris. Clearly in the case of V. corneae-infected E6 cells the parasite-host cell relationship involves subverting the host cell cytoskeleton and cell volume control, providing the parasite with the same protected niche as does a xenoma.

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Ultrastructural and immunohistological characterization of the sirc corneal cell line

Cited by 55 publications

References 17 publications

Antioxidant protection in cultured corneal cells and whole corneas submitted to UV-B exposure

Antioxidant protection in cultured corneal cells and whole corneas submitted to UV-B exposure

Functional differences in nucleoside and nucleobase transporters expressed on the rabbit corneal epithelial cell line (SIRC) and isolated rabbit cornea

Multinucleate host cells induced by Vittaforma corneae (Microsporidia)

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