Staining of Chlamydia trachomatis elementary bodies: a suitable method for identifying infected human monocytes by flow cytometry.
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.
Schnitger
Karen
K
Medizinische Hochschule Hannover, Department of Rheumatology, Hannover 30625, Germany. Schnitger.Karen@mh-hannover.de
Njau
Florence
F
Wittkop
Ulrike
U
Liese
Andrea
A
Kuipers
Jens G
JG
Thiel
Andreas
A
Morgan
Michael A
MA
Zeidler
Henning
H
Wagner
Annette D
AD
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
C087165:5-(6)-carboxyfluorescein diacetate succinimidyl ester; D005452:Fluoresceins; D005456:Fluorescent Dyes; D013388:Succinimides