SV40-immortalized and primary cultured human retinal pigment epithelial cells share similar patterns of cytokine-receptor expression and cytokine responsiveness

Brian D. Sippy Department of Pathology, University of Southern California School of Medicine, Los Angeles, CA, USA

Florence M. Hofman Department of Pathology, University of Southern California School of Medicine, Los Angeles, CA, USA; Department of Ophthalmology, University of Southern California School of Medicine, Los Angeles, CA, USA

Shikun He Department of Pathology, University of Southern California School of Medicine, Los Angeles, CA, USA

Roman Osusky Department of Ophthalmology, University of Southern California School of Medicine, Los Angeles, CA, USA

Shwu-Jiuan Sheu Department of Ophthalmology, University of Southern California School of Medicine, Los Angeles, CA, USA

Sharyn M. Walker Department of Microbiology, University of Southern California School of Medicine, Los Angeles, CA, USA; Childrens Hospital Los Angeles, Los Angeles, CA, USA

Stephen J. Ryan Department of Ophthalmology, University of Southern California School of Medicine, Los Angeles, CA, USA; Doheny Eye Institute, Los Angeles, CA, USA

David R. Hinton Department of Pathology, University of Southern California School of Medicine, Los Angeles, CA, USA; Department of Neurology, University of Southern California School of Medicine, Los Angeles, CA, USA; Department of Neurological Surgery, University of Southern California School of Medicine, Los Angeles, CA, USA

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Abstract

Retinal pigment epithelial (RPE) cells produce and respond to a variety of cytokines; however, molecular and biochemical studies are restricted by the limited access to large numbers of pure cells and the variability associated with different donor sources. Despite success in establishing primary human RPE (HRPE) cell cultures, the inability to sustain consistent proliferation rates and morphology over several passages remains a concern. This problem was approached by using an immortalized line of simian virus (SV)40 transformed fetal HRPE cells (SVRPE). Cytokine production, receptor expression and responsiveness in the SVRPE cell line was analyzed to determine the usefulness of this model for studying HRPE-cytokine interactions. Using reverse transcriptase polymerase chain reaction (RT-PCR), HRPE and SVRPE cells demonstrated an identical pattern of interleukin-1 receptor (IL-1R), IL-2R (α sub-unit), IL-6R, interferon (IFN)-γR and tumor necrosis factor-α (TNF)R p55 expression. No amplification products for TNFR p75 or granulocyte/macrophage colony stimulating factor (GM-CSF)R were demonstrated in either population. IFN-γ stimulation induced surface human leukocyte antigen (HLA)-DR in both SVRPE and HRPE, while TNF treatment induced surface expression of intercellular adhesion molecule (ICAM)-1 on SVRPE and upregulated ICAM from basal levels on HRPE. Both cell types showed amplification products for interleukin (IL)-1 β, IL-6 and transforming growth factor (TGF)-βl using RT-PCR. The bioassays demonstrated that both populations of unstimulated cells constitutively secrete very low levels of TGF-β and no IL-6. The results indicate that SVRPE cells express cytokine-receptors, respond to cytokine stimulation, and produce cytokines similar to that of primary cultured HRPE cells. Thus, this functionally and morphologically consistent SVRPE cell line represents an excellent model for investigating the cellular and molecular aspects of human RPE-cytokine regulation.

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SV40-immortalized and primary cultured human retinal pigment epithelial cells share similar patterns of cytokine-receptor expression and cytokine responsiveness

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SV40-immortalized and primary cultured human retinal pigment epithelial cells share similar patterns of cytokine-receptor expression and cytokine responsiveness

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