A platform for studying the transfer of Chlamydia pneumoniae infection between respiratory epithelium and phagocytes

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http://hdl.handle.net/10138/313183

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Kortesoja , M , Trofin , R E & Hanski , L 2020 , ' A platform for studying the transfer of Chlamydia pneumoniae infection between respiratory epithelium and phagocytes ' , Journal of Microbiological Methods , vol. 171 , 105857 . https://doi.org/10.1016/j.mimet.2020.105857

Title: A platform for studying the transfer of Chlamydia pneumoniae infection between respiratory epithelium and phagocytes
Author: Kortesoja, Maarit; Trofin, Raluca Elena; Hanski, Leena
Contributor: University of Helsinki, Pharmaceutical Design and Discovery group
University of Helsinki, Division of Pharmaceutical Biosciences
Date: 2020-04
Language: eng
Number of pages: 9
Belongs to series: Journal of Microbiological Methods
ISSN: 0167-7012
URI: http://hdl.handle.net/10138/313183
Abstract: 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.
Subject: Intracellular bacterium
Coculture
Persistent infection
Dissemination
HLAMYDOPHILAC-PNEUMONIAE
SYSTEMIC DISSEMINATION
HUMAN MONOCYTES
CELL-LINE
HL CELLS
ACTIVATION
IDENTIFICATION
FLUORESCENCE
MACROPHAGES
3111 Biomedicine
318 Medical biotechnology
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