Browsing by Subject "CELL-LINE"

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  • Kortesoja, Maarit; Trofin, Raluca Elena; Hanski, Leena (2020)
    The obligate intracellular bacterium, Chlamydia pneumoniae, has been identified as a risk factor for several chronic inflammatory diseases in addition to respiratory tract infections. The dissemination of C. pneumoniae from respiratory tract to secondary sites of infection occurs via infected monocyte / macrophage line cells, in which C. pneumoniae can persist as an antibiotic-refractory phenotype. To allow more detailed studies on the epithelium-monocyte/macrophage transition of the infection, new in vitro bioassays are needed. To this end, a coculture system with human continuous cell lines was established. Respiratory epithelial HL cells were infected with C. pneumoniae and THP-1 monocytes were added into the cultures at 67 h post infection. After a 5 h coculture, THP-1 cells were collected with a biotinylated HLA antibody and streptavidin-coated magnetic beads and C. pneumoniae genome copy numbers in THP-1 determined by quantitative PCR. The assay was optimized for cell densities, incubation time, THP-1 separation technique and buffer composition, and its robustness was demonstrated by a Z' value of 0.6. The mitogen-activated protein kinase (MAPK) inhibitors: SP600125 (JNK inhibitor), SB203580 (p38 inhibitor) and FR180204 (ERK inhibitor) suppressed the transfer of C. pneumoniae from HL to THP-1 cells, making them suitable positive controls for the assay. Based on analysis of separate steps of the process, the MAPK inhibitors suppress the bacterial entry to THP-1 cells. The transfer of C. pneumoniae from epithelium to phagocytes represents a crucial step in the establishment of persistent infections by this pathogen, and the presented methods enables future studies to block this process by therapeutic means.
  • Jokela, Jouni; Oftedal, Linn; Herfindal, Lars; Permi, Perttu; Wahlsten, Matti; Doskeland, Stein Ove; Sivonen, Kaarina (2012)
  • Abu Saleh, Doaa; Niskanen, Jukka; Xue, Yanming; Golberg, Dmitri; Winnik, Francoise M.; Sosnik, Alejandro (2017)
    We report an organic-inorganic hybrid core-shell nanomaterial obtained by conjugation of an amphiphilic monomethoxy-poly(ethylene glycol)-b-poly(epsilon-caprolactone) diblock copolymer to hydroxylated boron nitride nanotubes (BNNTs). The extent of copolymer grafting reached 64% w/w, an exceptionally high value. The hybrid materials exhibit excellent physical stability in water and an outstanding loading capacity (31.3% w/w) for curcumin, a hydrophobic drug. Moreover, they present good compatibility with the Caco2 cell line, a model of intestinal epithelium. Our findings demonstrate the potential of multifunctional hybrid BNNTs to serve as a platform for complex amphiphilic nanoparticle architectures with improved features. (c) 2017 Elsevier Ltd. All rights reserved.
  • Prusty, Bhupesh K.; Siegl, Christine; Hauck, Petra; Hain, Johannes; Korhonen, Suvi J.; Hiltunen-Back, Eija; Puolakkainen, Mirja; Rudel, Thomas (2013)
  • Sadeghi, Mohammadreza; Popov, Vsevolod; Guzman, Hilda; Phan, Tung Gia; Vasilakis, Nikos; Tesh, Robert; Delwart, Eric (2017)
    Eleven viral isolates derived mostly in albopictus C6/36 cells from mosquito pools collected in Southeast Asia and the Americas between 1966 and 2014 contained particles with electron microscopy morphology typical of reoviruses. Metagenomics analysis yielded the near complete genomes of three novel reoviruses, Big Cypress orbivirus, Ninarumi virus, and High Island virus and a new tetravirus, Sarawak virus. Strains of previously characterized Sathuvarachi, Yunnan, Banna and Parry's Lagoon viruses (Reoviridae), Bontang virus (Mesoniviridae), and Culex theileri flavivirus (Flaviviridae) were also characterized. The availability of these mosquito virus genomes will facilitate their detection by metagenomics or PCR to better determine their geographic range, extent of host tropism, and possible association with arthropod or vertebrate disease.
  • Kurppa, Kari J.; Caton, Javier; Morgan, Peter R.; Ristimaki, Ari; Ruhin, Blandine; Kellokoski, Jari; Elenius, Klaus; Heikinheimo, Kristiina (2014)
  • Jaumotte, Juliann D.; Saarma, Mart; Zigmond, Michael J. (2021)
    Parkinson's disease is associated with the loss of dopamine (DA) neurons in ventral mesencephalon. We have previously reported that no single neurotrophic factor we tested protected DA neurons from the dopaminergic toxin 1-methyl-4-phenylpyridinium (MPP+) in dissociated cultures isolated from the P0 rat substantia nigra, but that a combination of five neurotrophic factors was protective. We now report that cerebral DA neurotrophic factor (CDNF) and a variant of neurturin (NRTN), N4, were also not protective when provided alone but were protective when added together. In cultures isolated from the substantia nigra, MPP+ (10 mu M) decreased tyrosine hydroxylase-positive cells to 41.7 +/- 5.4% of vehicle control. Although treatment of cultures with 100 ng/ml of either CDNF or N4 individually before and after toxin exposure did not significantly increase survival in MPP+-treated cultures, when the two trophic factors were added together at 100 ng/ml each, survival of cells was increased 28.2 +/- 6.1% above the effect of MPP+ alone. In cultures isolated from the ventral tegmental area, another DA rich area, a higher dose of MPP+ (1 mM) was required to produce an EC50 in TH-positive cells but, as in the substantia nigra, only the combination of CDNF and N4 (100 ng/ml each) was successful at increasing the survival of these cells compared to MPP+ alone (by 22.5 +/- 3.5%). These data support previous findings that CDNF and N4 may be of therapeutic value for treatment of PD, but suggest that they may need to be administered together.