Mary Polan
Publication Details
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Interleukin 1 beta regulates Vero cell interleukin-1 receptor type I messenger ribonucleic acid expression.
Biol Reprod. 1997; (4): 783-90
Embryos cocultured with Vero cells display enhanced development in vitro. This could be due to an interaction between the embryo and cellular layer mediated by paracrine cytokines. The interleukin-1 (IL-1) system is composed of two IL-1 agonists, IL-1 receptor antagonist (IL-1ra), and two major IL-1 receptors (IL-1R tII). In this study, we measured Vero cell expression of IL-1 system mRNAs with reverse transcription polymerase chain reaction (RT-PCR) and validated the results with an immunohistochemistry study. RT-PCR revealed beta-actin and IL-1R tI mRNA expression in Vero cell cocultures without detectable IL-1 beta and IL-1ra mRNA expression. To determine the effect of IL-1 beta on IL-1R tI mRNA expression in Vero cells, quantitative competitive PCR methodology was developed. A competitive cDNA fragment was coamplified as an internal standard with the target cDNA sequence of IL-1R tI, showing a 50% decrease in Vero cell IL-1R tI cDNA cultured in the presence of IL-1 beta (100 IU/mI) compared to control Vero cell cultures (62.5 fg vs. 125 fg). Down-regulation of IL-1R tI mRNA by IL-1 beta is dose-dependent, with increasing concentrations (0-1000) IU/ml) of IL-1 beta producing progressive attenuation of IL-1R tI expression. Treatment with anti-IL-1 beta antibody ablate the inhibitory effect of IL-1 beta (100 IU/ml) on IL-1R tI mRNA expression. Immunostaining confirmed the presence of IL-1r tI protein in Vero cells. These results demonstrate the presence of IL-1 R tI in Vero cell monolayers and regulation of Il-1R tI mRNA by IL-1 beta.
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