Contact of Chlamydophila pneumoniae with type II cell triggers activation of calcium-mediated NF-kappa B pathway.
Nuclear factor-kappa B (NF-kappa B) plays an important role in inflammation, proliferation and regulation of apoptosis. The purpose of the present study on type II cells was to investigate whether Chlamydophila pneumoniae contact induces (I) a Ca2+ release, that (II) disrupts F-actin/beta-tubulin cytoskeletal association with NF-kappa B/I kappa B alpha, leading to (III) a subsequent NF-kappa B activation. Incubation of rat type II pneumocytes with C. pneumoniae caused an intracellular calcium release within seconds. Confocal laser scanning microscopy (CLSM) revealed that bacterial contact with cell surface leads to a disappearance of the microvilli and disturbs the co-localization between F-actin and NF-kappa B (p65). Using semi-quantitative CLSM, we show that at 10-30 min I kappa B alpha was decreased and p65 or p50 was simultaneously translocated from cytoplasm to the nucleus, resulting in a 19-fold and 17-fold increase versus control cells. During this time no bacteria were internalized into type II cells. The pre-treatment of cells with BAPTA-AM inhibited C. pneumoniae-mediated calcium release. BAPTA-AM or SN50 prevented the C. pneumoniae-induced changes in F-actin cytoskeleton and inhibited NF-kappa B activation. Paclitaxel reduced C. pneumoniae-mediated changes of beta-tubulin cytoskeleton and activation of NF-kappa B. These results suggest that calcium-mediated cytoskeleton reorganization is involved in C. pneumoniae-induced NF-kappa B activation in type II cells.
Wissel
Heide
H
Clinic of Neonatology, Campus Charité-Mitte, University Children's Hospital, University Medicine Berlin, Germany. hwissel@charite.de
Müller
Torsten
T
Rüdiger
Mario
M
Krüll
Matthias
M
Wauer
Roland R
RR
D000199:Actins / Q000378:metabolism; D000818:Animals; D015153:Blotting, Western; D002118:Calcium / Q000378:metabolism; D002467:Cell Nucleus / Q000378:metabolism; D002478:Cells, Cultured; D016993:Chlamydophila pneumoniae / Q000378:metabolism*; D003593:Cytoplasm / Q000378:metabolism; D003599:Cytoskeleton / Q000378:metabolism; D004533:Egtazic Acid / Q000031:analogs & derivatives* / Q000494:pharmacology; D007150:Immunohistochemistry; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008168:Lung / Q000166:cytology* / Q000382:microbiology*; D008297:Male; D018613:Microscopy, Confocal; D016328:NF-kappa B / Q000378:metabolism*; D011485:Protein Binding; D021381:Protein Transport; D051381:Rats; D017208:Rats, Wistar; D013997:Time Factors; D014404:Tubulin / Q000378:metabolism
D000199:Actins; D016328:NF-kappa B; D014404:Tubulin; C070379:1,2-bis(2-aminophenoxy)ethane N,N,N',N'-tetraacetic acid acetoxymethyl ester; D004533:Egtazic Acid; D007770:L-Lactate Dehydrogenase; D002118:Calcium