Interaction of Bartonella henselae with endothelial cells results in rapid bacterial rRNA synthesis and replication.
Bartonella henselae is a slow-growing microorganism and the causative pathogen of bacillary angiomatosis in man. Here, we analysed how interaction of B. henselae with endothelial cells might affect bacterial growth. For this purpose, bacterial rRNA production and ribosome content was determined by fluorescence in situ hybridization (FISH) using rRNA-targeted fluorescence-labelled oligonucleotide probes. B. henselae grown on agar plates showed no detectable rRNA content by means of FISH, whereas B. henselae co-cultured with endothelial cells showed a rapid increase of rRNA production within the first 18 h after inoculation. The increased rRNA synthesis was paralleled by a approximately 1000-fold intracellular bacterial replication, whereas bacteria grown on agar base showed only a approximately 10-fold replication within the first 48 h of culture. Pretreatment of host cells with paraformaldehyde prevented adhesion, invasion, intracellular replication and bacterial rRNA synthesis of B. henselae. In contrast, inhibition of host cell protein synthesis by cycloheximide did not affect bacterial adhesion and invasion, but prevented intracellular replication although bacterial rRNA content was increased. Inhibition of actin polymerization by cytochalasin D did not affect adhesion, invasion, increased rRNA content or intracellular replication of B. henselae. These results demonstrate that rRNA synthesis and replication of B. henselae is promoted by viable host cells with intact de novo protein synthesis.
Kempf
V A
VA
Max von Pettenkofer-Institut für Hygiene und Medizinische Mikrobiologie, Ludwig Maximilians Universität München, Munich, Germany. kempf@m3401.mpk.med.uni-muenchen.de
Schaller
M
M
Behrendt
S
S
Volkmann
B
B
Aepfelbacher
M
M
Cakman
I
I
Autenrieth
I B
IB
D000900:Anti-Bacterial Agents / Q000494:pharmacology; D018416:Bartonella henselae / Q000235:genetics / Q000472:pathogenicity*; D002460:Cell Line; D003513:Cycloheximide / Q000494:pharmacology; D015638:Cytochalasin D / Q000494:pharmacology; D004727:Endothelium / Q000187:drug effects / Q000382:microbiology*; D005557:Formaldehyde / Q000494:pharmacology; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D017348:Microscopy, Electron, Scanning Transmission; D019384:Nucleic Acid Synthesis Inhibitors / Q000494:pharmacology; D011108:Polymers / Q000494:pharmacology; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D012329:RNA, Bacterial / Q000032:analysis; D012335:RNA, Ribosomal / Q000032:analysis
D000900:Anti-Bacterial Agents; D019384:Nucleic Acid Synthesis Inhibitors; D011108:Polymers; D011500:Protein Synthesis Inhibitors; D012329:RNA, Bacterial; D012335:RNA, Ribosomal; D005557:Formaldehyde; D015638:Cytochalasin D; D003513:Cycloheximide; C003043:paraform