Mervi S Riekkinen

University of Kuopio, Kuopio, Eastern Finland Province, Finland

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Publications (5)20.04 Total impact

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    ABSTRACT: Adenoviral gene therapy, based on herpes simplex virus thymidine kinase (AdvTK) is being developed for clinical use but no safety data are available with respect to the effects on female germ cells should the virus accidentally be released into systemic circulation. We studied the effects of AdvTK gene therapy on ovaries and germ cells in pregnant and nonpregnant rabbits and also the potential transmission of a transgene to offspring. To mimic the severest of conditions, gene transfer was made by direct catheter-mediated injection into the uterine artery of pregnant and nonpregnant rabbits. AdvTK or AdvLacZ at 1 x 10(10) pfu were used for gene transfer. Ganciclovir was administered to AdvTK-treated rabbits to induce gene therapy. The rabbits were mated 6 and 12 weeks following gene transfer and the surviving young (89 from a total of 114) were analysed. No change in fertility was observed in the two matings after the gene transfer. In addition, no change was observed in ovarian histology between the AdvTK group, the AdvLacZ group and the nontreated controls. Southern blotting analysis showed no genomic integration of the transgene. However, in PCR analysis, transgene DNA was found in 9.3% of the litter samples. This was not the case for results from the reverse transcription-PCR assay. Although AdvTK gene therapy may initially affect ovarian cells, the influence appears to be transient. However, after direct exposure of the ovarian cells in high concentration of adenoviruses, transmission of a transgene in the offspring cannot be excluded.
    No preview · Article · Sep 2008 · The Journal of Gene Medicine
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    ABSTRACT: Baculoviruses can express transgenes in a wide range of vertebrate cells. However, in some cells transgene expression is weak. To enhance transgene expression, we studied the effect of the Woodchuck hepatitis virus post-transcriptional regulatory element (WPRE) on baculovirus (BV)-mediated gene expression of several transgenes. A significant increase in BV-mediated gene expression was detected in several cell lines. A 10-fold increase in transgene expression was observed with the WPRE as determined by the percentage of positive cells and mean fluorescence intensity (MFI). Furthermore, a combination of optimized cell culture medium and WPRE virus led to more than a 60-fold increase in gene expression. In accordance, elevated mRNA and protein levels were detected in WPRE-virus transduced cells. In HepG2 and RaaSMC, WPRE-mediated enhancement was comparable to the previously shown positive effect of sodium butyrate on BV-mediated gene expression. Thus, inclusion of the WPRE into a baculovirus vector provides a simple means to improve BV-mediated gene expression in vertebrate cells.
    No preview · Article · Sep 2007 · Journal of Biotechnology
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    ABSTRACT: To better understand the role of macrophages in atherogenesis and to find new strategies to prevent their harmful effects, more information is needed about their gene and protein expression patterns in atherogenic conditions. We analyzed gene and protein expression changes during monocyte-macrophage differentiation and lipid-loading by cDNA arrays and antibody-based protein arrays, respectively. It was found that early response genes, such as transcription factors, were upregulated early during monocyte-macrophage differentiation, while genes functioning in cell proliferation, migration, inflammation and lipid metabolism were activated later during macrophage differentiation. When comparing results from cDNA and antibody arrays, it become evident that changes at the protein levels were not always predicted by changes at the mRNA level. This discrepancy may be due to the different transcript variants that exist for distinct genes, posttranslational modifications and different turnover rates for mRNAs and proteins of distinct genes. When combining cDNA and protein array results with RT-PCR, it was found that CD36, COX-2, and several factors regulating cell signaling, such as Cdk-1, TFII-I, NEMO-like kinase, Elf-5 and TRADD were strongly upregulated both at the mRNA and protein levels. Time-dependency of the activation of the early response genes and genes functioning in inflammation, lipid metabolism and cell proliferation and migration, is an important feature of the macrophage differentiation. It was also evident that several novel transcription factors were activated during lipid-loading. It is concluded that cDNA and protein arrays will be useful for the identification of genes that are potential targets for therapeutic interventions.
    Full-text · Article · Jul 2005 · Atherosclerosis
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    ABSTRACT: In utero gene therapy has a potential to correct genetic disorders before the first clinical symptoms appear. Our aim was to examine whether the exocoelomic cavity between amniotic and chorionic membranes offers a minimally invasive route for gene transfer to the fetus during early pregnancy. We injected lacZ-adenovirus (4 x 10(9) pfu) during open surgery into the exocoelomic cavity of rat fetuses (n=50) and analyzed the fetuses and rat dams for transgene expression with X-gal staining and polymerase chain reaction. Giant cells around Reichert's membrane, the outermost extraembryonic membrane in rodents, were transduced; but no transduction was observed in the cells of the fetuses or rat dams. In rodents, the exocoelomic cavity does not offer a route for gene transfer into the fetus. It was concluded that fetal membranes act as a barrier that prevents adenoviral particles from passing between embryonic cavities.
    No preview · Article · Feb 2004 · American Journal of Obstetrics and Gynecology
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    ABSTRACT: Inflammatory cells play an important role in atherogenesis. However, more information is needed about their gene expression profiles in human lesions. We used laser microdissection (LMD) to isolate macrophage-rich shoulder areas from human lesions. Gene expression profiles in isolated cells were analyzed by cDNA array and compared with expression patterns in normal intima and THP-1 macrophages. Upregulation of 72 genes was detected with LMD and included 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase, interferon regulatory factor-5 (IRF-5), colony stimulating factor (CSF) receptors, CD11a/CD18 integrins, interleukin receptors, CD43, calmodulin, nitric oxide synthase (NOS), and extracellular superoxide dismutase (SOD). Several of these changes were also present in PMA-stimulated THP-1 macrophages in vitro. On the other hand, expression of several genes, such as VEGF, tissue factor pathway inhibitor 2, and apolipoproteins C-I and C-II, decreased. Overexpression of HMG-CoA reductase in macrophage-rich lesion areas may explain some beneficial effects of statins, which can also modulate increased expression of CD11a/CD18 and CD43 found in microdissected cells. We also found increased expression of CSF receptors, IRF-5, and interleukin receptors, which could become useful therapeutic targets for the treatment of atherosclerotic diseases.
    Full-text · Article · Jan 2004 · Arteriosclerosis Thrombosis and Vascular Biology

Publication Stats

150 Citations
20.04 Total Impact Points


  • 2004-2008
    • University of Kuopio
      • Department of Medicine
      Kuopio, Eastern Finland Province, Finland
  • 2007
    • Institute for Molecular Medicine, Finland
      Helsinki, Southern Finland Province, Finland