title: Staining of Chlamydia trachomatis elementary bodies: a suitable method for identifying infected human monocytes by flow cytometry.
abstract: Persistence of Chlamydia trachomatis (C. trachomatis) in the joint is the most frequent cause of reactive arthritis following urogenital tract infection. The resulting changes of host cell antigen- and cytokine-expression are not precisely understood. We developed and evaluated a direct cytometric approach to visualize in vitro C. trachomatis-infected monocytes. Infectious elementary bodies (EBs) of C. trachomatis serovar K were labelled by incubation with 5-(and-6)-carboxyfluorescein diacetate succinimidyl ester (CFSE). Afterwards, human peripheral blood monocytes were cultured with the CFSE-labelled EBs and analysed by flow cytometry. Real-time polymerase chain reaction (PCR) was used to demonstrate intracellular uptake and viability of CFSE-labelled C. trachomatis by the determination of gene expression. Labelling EBs with CFSE may become a valuable tool for studying the interaction between C. trachomatis and the host cell.
authors.affiliation: Medizinische Hochschule Hannover, Department of Rheumatology, Hannover 30625, Germany. Schnitger.Karen@mh-hannover.de
authors.forename: Karen
authors.initials: K
authors.lastname: Schnitger
authors.forename: Florence
authors.initials: F
authors.lastname: Njau
authors.forename: Ulrike
authors.initials: U
authors.lastname: Wittkop
authors.forename: Andrea
authors.initials: A
authors.lastname: Liese
authors.forename: Jens G
authors.initials: JG
authors.lastname: Kuipers
authors.forename: Andreas
authors.initials: A
authors.lastname: Thiel
authors.forename: Michael A
authors.initials: MA
authors.lastname: Morgan
authors.forename: Henning
authors.initials: H
authors.lastname: Zeidler
authors.forename: Annette D
authors.initials: AD
authors.lastname: Wagner
mesh_terms: D002478:Cells, Cultured; D002692:Chlamydia trachomatis / Q000302:isolation & purification* / Q000648:ultrastructure; D005434:Flow Cytometry / Q000379:methods*; D005452:Fluoresceins / Q000737:chemistry; D005456:Fluorescent Dyes / Q000737:chemistry; D006801:Humans; D009000:Monocytes / Q000382:microbiology*; D013388:Succinimides / Q000737:chemistry; D013997:Time Factors
chemical_list: C087165:5-(6)-carboxyfluorescein diacetate succinimidyl ester; D005452:Fluoresceins; D005456:Fluorescent Dyes; D013388:Succinimides