SV40-IMMORTALIZED AND PRIMARY CULTURED HUMAN RETINAL-PIGMENT EPITHELIAL-CELLS SHARE SIMILAR PATTERNS OF CYTOKINE-RECEPTOR EXPRESSION AND CYTOKINE RESPONSIVENESS

被引：34

作者：

SIPPY, BD

HOFMAN, FM

HE, SK

OSUSKY, R

SHEU, SJ

WALKER, SM

RYAN, SJ

HINTON, DR

机构：

[1] UNIV SO CALIF, SCH MED, DEPT PATHOL, LOS ANGELES, CA 90033 USA

[2] UNIV SO CALIF, SCH MED, DEPT OPHTHALMOL, LOS ANGELES, CA 90033 USA

[3] UNIV SO CALIF, SCH MED, DEPT MICROBIOL, LOS ANGELES, CA 90033 USA

[4] UNIV SO CALIF, SCH MED, DEPT NEUROL, LOS ANGELES, CA 90033 USA

[5] UNIV SO CALIF, SCH MED, DEPT NEUROL SURG, LOS ANGELES, CA 90033 USA

[6] CHILDRENS HOSP LOS ANGELES, LOS ANGELES, CA USA

[7] DOHENY EYE INST, LOS ANGELES, CA USA

来源：

CURRENT EYE RESEARCH | 1995年 / 14卷 / 06期

关键词：

CELL CULTURE; INTERCELLULAR ADHESION MOLECULE (ICAM)-1; SIMIAN VIRUS (SV)-40; CYTOKINES; REVERSE TRANSCRIPTASE-POLYMERASE CHAIN REACTION (RT-PCR); TUMOR NECROSIS FACTOR-ALPHA (TNF-ALPHA); HUMAN; RETINAL PIGMENT EPITHELIUM (RPE);

D O I：

10.3109/02713689509003761

中图分类号：

R77 [眼科学];

学科分类号：

100212 ;

摘要：

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 (alpha sub-unit), IL-6R, interferon (IFN)-gamma R and tumor necrosis factor-alpha (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-gamma 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 beta, IL-6 and transforming growth factor (TGF)-beta 1 using RT-PCR. The bioassays demonstrated that both populations of unstimulated cells constitutively secrete very low levels of TGF-beta 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.

引用

页码：495 / 503

页数：9

共 42 条

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