Fluorescence lifetime imaging unravels C. trachomatis metabolism and its crosstalk with the host cell

Márta Szaszák, Philipp Steven, Kensuke Shima, Regina Orzekowsky-Schröder, Gereon Hüttmann, Inke R. König, Werner Solbach, Jan Rupp*

*Corresponding author for this work
23 Citations (Scopus)

Abstract

Chlamydia trachomatis is an obligate intracellular bacterium that alternates between two metabolically different developmental forms. We performed fluorescence lifetime imaging (FLIM) of the metabolic coenzymes, reduced nicotinamide adenine dinucleotides [NAD(P)H], by two-photon microscopy for separate analysis of host and pathogen metabolism during intracellular chlamydial infections. NAD(P)H autofluorescence was detected inside the chlamydial inclusion and showed enhanced signal intensity on the inclusion membrane as demonstrated by the co-localization with the 14-3-3β host cell protein. An increase of the fluorescence lifetime of protein-bound NAD(P)H [τ 2-NAD(P)H] inside the chlamydial inclusion strongly correlated with enhanced metabolic activity of chlamydial reticulate bodies during the mid-phase of infection. Inhibition of host cell metabolism that resulted in aberrant intracellular chlamydial inclusion morphology completely abrogated the τ 2-NAD(P)H increase inside the chlamydial inclusion. τ 2-NAD(P)H also decreased inside chlamydial inclusions when the cells were treated with IFNγ reflecting the reduced metabolism of persistent chlamydiae. Furthermore, a significant increase in τ 2-NAD(P)H and a decrease in the relative amount of free NAD(P)H inside the host cell nucleus indicated cellular starvation during intracellular chlamydial infection. Using FLIM analysis by two-photon microscopy we could visualize for the first time metabolic pathogen-host interactions during intracellular Chlamydia trachomatis infections with high spatial and temporal resolution in living cells. Our findings suggest that intracellular chlamydial metabolism is directly linked to cellular NAD(P)H signaling pathways that are involved in host cell survival and longevity.

Original languageEnglish
Article numbere1002108
JournalPLoS Pathogens
Volume7
Issue number7
ISSN1553-7366
DOIs
Publication statusPublished - 01.07.2011

Research Areas and Centers

  • Academic Focus: Center for Infection and Inflammation Research (ZIEL)

Fingerprint

Dive into the research topics of 'Fluorescence lifetime imaging unravels C. trachomatis metabolism and its crosstalk with the host cell'. Together they form a unique fingerprint.

Cite this