title: Interleukin 1 beta regulates Vero cell interleukin-1 receptor type I messenger ribonucleic acid expression.
abstract: 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.
authors.affiliation: Department of Gynecology and Obstetrics, Stanford University Center and School of Medicine, California 94305, USA.
authors.forename: H Y
authors.initials: HY
authors.lastname: Huang
authors.forename: Y
authors.initials: Y
authors.lastname: Wen
authors.forename: D
authors.initials: D
authors.lastname: Valbuena
authors.forename: J S
authors.initials: JS
authors.lastname: Krüssel
authors.forename: M L
authors.initials: ML
authors.lastname: Polan
mesh_terms: D000818:Animals; D002522:Chlorocebus aethiops; D017931:DNA Primers; D015536:Down-Regulation / Q000187:drug effects; D004587:Electrophoresis, Agar Gel; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007150:Immunohistochemistry; D007375:Interleukin-1 / Q000502:physiology*; D015345:Oligonucleotide Probes; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000096:biosynthesis*; D017472:Receptors, Interleukin-1 / Q000096:biosynthesis*; D011994:Recombinant Proteins / Q000096:biosynthesis; D014158:Transcription, Genetic; D014709:Vero Cells
chemical_list: D017931:DNA Primers; D007375:Interleukin-1; D015345:Oligonucleotide Probes; D012333:RNA, Messenger; D017472:Receptors, Interleukin-1; D011994:Recombinant Proteins