title: Chlamydia pneumoniae growth inhibition in human monocytic THP-1 cells and human epithelial HEp-2 cells by a novel phenoxazine derivative.
abstract: In this study the effects of 2-amino-phenoxazine-3-one (phenoxazine derivate, Phx-3) on Chlamydia (Chlamydophila) pneumoniae growth in human monocytic THP-1 cells as well as human epithelial HEp-2 cells were examined. Cells were infected with bacteria at an m.o.i. of 10 by centrifugation. After washing to remove any remaining bacteria, the cells were incubated with or without Phx-3 in the presence or absence of tryptophan for 72 h. The bacteria in cells were assessed by staining of chlamydial inclusions with FITC-labelled anti-chlamydial antibody, electron microscopic analysis, real-time RT-PCR specific for C. pneumoniae 16S rRNA and propagation on HEp-2 cells. Treatment with Phx-3 significantly inhibited growth of C. pneumoniae in THP-1 and HEp-2 cells. A decrease in the number of bacterial 16S rRNA transcripts was also confirmed in both cell lines by real-time RT-PCR. Electron microscopic studies revealed that treatment with Phx-3 induces bacterial destruction in most of the inclusion bodies in these cells. Addition of tryptophan to the culture slightly blocked the growth inhibition of C. pneumoniae by Phx-3. The reagents did not show any cytotoxicity to the cells at the concentrations used. The results suggest that Phx-3 inhibits C. pneumoniae replication in human monocytic cells as well as epithelial cells, partially depending on the tryptophan-metabolic pathway of host cells. Thus, Phx-3 might be a useful compound for controlling C. pneumoniae growth in cells and may be an alternative conventional therapy.
authors.affiliation: Department of Infectious Disease, Division of Microbiology1, First Department of Internal Medicine2, Laboratory of Electron Microscopy4 and Department of Anatomy5, Kyorin University School of Medicine,Tokyo 181-8611, Japan 3Division of Molecular Microbiology, Department of Basic Laboratory Sciences, Graduate School of Medicine, Osaka University, 1-7 Yamadaoka, Suita, Osaka 565-0871, Japan 6Department of Virology I, National Institute of Infectious Diseases, Tokyo 162-8642, Japan 7Department of Biochemistry, Tokyo Medical University, Tokyo 160-0022, Japan.
authors.forename: Tomonori
authors.initials: T
authors.lastname: Uruma
authors.affiliation: Department of Infectious Disease, Division of Microbiology1, First Department of Internal Medicine2, Laboratory of Electron Microscopy4 and Department of Anatomy5, Kyorin University School of Medicine,Tokyo 181-8611, Japan 3Division of Molecular Microbiology, Department of Basic Laboratory Sciences, Graduate School of Medicine, Osaka University, 1-7 Yamadaoka, Suita, Osaka 565-0871, Japan 6Department of Virology I, National Institute of Infectious Diseases, Tokyo 162-8642, Japan 7Department of Biochemistry, Tokyo Medical University, Tokyo 160-0022, Japan.
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Yamaguchi
authors.affiliation: Department of Infectious Disease, Division of Microbiology1, First Department of Internal Medicine2, Laboratory of Electron Microscopy4 and Department of Anatomy5, Kyorin University School of Medicine,Tokyo 181-8611, Japan 3Division of Molecular Microbiology, Department of Basic Laboratory Sciences, Graduate School of Medicine, Osaka University, 1-7 Yamadaoka, Suita, Osaka 565-0871, Japan 6Department of Virology I, National Institute of Infectious Diseases, Tokyo 162-8642, Japan 7Department of Biochemistry, Tokyo Medical University, Tokyo 160-0022, Japan.
authors.forename: Minoru
authors.initials: M
authors.lastname: Fukuda
authors.affiliation: Department of Infectious Disease, Division of Microbiology1, First Department of Internal Medicine2, Laboratory of Electron Microscopy4 and Department of Anatomy5, Kyorin University School of Medicine,Tokyo 181-8611, Japan 3Division of Molecular Microbiology, Department of Basic Laboratory Sciences, Graduate School of Medicine, Osaka University, 1-7 Yamadaoka, Suita, Osaka 565-0871, Japan 6Department of Virology I, National Institute of Infectious Diseases, Tokyo 162-8642, Japan 7Department of Biochemistry, Tokyo Medical University, Tokyo 160-0022, Japan.
authors.forename: Hayato
authors.initials: H
authors.lastname: Kawakami
authors.affiliation: Department of Infectious Disease, Division of Microbiology1, First Department of Internal Medicine2, Laboratory of Electron Microscopy4 and Department of Anatomy5, Kyorin University School of Medicine,Tokyo 181-8611, Japan 3Division of Molecular Microbiology, Department of Basic Laboratory Sciences, Graduate School of Medicine, Osaka University, 1-7 Yamadaoka, Suita, Osaka 565-0871, Japan 6Department of Virology I, National Institute of Infectious Diseases, Tokyo 162-8642, Japan 7Department of Biochemistry, Tokyo Medical University, Tokyo 160-0022, Japan.
authors.forename: Hajime
authors.initials: H
authors.lastname: Goto
authors.affiliation: Department of Infectious Disease, Division of Microbiology1, First Department of Internal Medicine2, Laboratory of Electron Microscopy4 and Department of Anatomy5, Kyorin University School of Medicine,Tokyo 181-8611, Japan 3Division of Molecular Microbiology, Department of Basic Laboratory Sciences, Graduate School of Medicine, Osaka University, 1-7 Yamadaoka, Suita, Osaka 565-0871, Japan 6Department of Virology I, National Institute of Infectious Diseases, Tokyo 162-8642, Japan 7Department of Biochemistry, Tokyo Medical University, Tokyo 160-0022, Japan.
authors.forename: Toshio
authors.initials: T
authors.lastname: Kishimoto
authors.affiliation: Department of Infectious Disease, Division of Microbiology1, First Department of Internal Medicine2, Laboratory of Electron Microscopy4 and Department of Anatomy5, Kyorin University School of Medicine,Tokyo 181-8611, Japan 3Division of Molecular Microbiology, Department of Basic Laboratory Sciences, Graduate School of Medicine, Osaka University, 1-7 Yamadaoka, Suita, Osaka 565-0871, Japan 6Department of Virology I, National Institute of Infectious Diseases, Tokyo 162-8642, Japan 7Department of Biochemistry, Tokyo Medical University, Tokyo 160-0022, Japan.
authors.forename: Yoshimasa
authors.initials: Y
authors.lastname: Yamamoto
authors.affiliation: Department of Infectious Disease, Division of Microbiology1, First Department of Internal Medicine2, Laboratory of Electron Microscopy4 and Department of Anatomy5, Kyorin University School of Medicine,Tokyo 181-8611, Japan 3Division of Molecular Microbiology, Department of Basic Laboratory Sciences, Graduate School of Medicine, Osaka University, 1-7 Yamadaoka, Suita, Osaka 565-0871, Japan 6Department of Virology I, National Institute of Infectious Diseases, Tokyo 162-8642, Japan 7Department of Biochemistry, Tokyo Medical University, Tokyo 160-0022, Japan.
authors.forename: Akio
authors.initials: A
authors.lastname: Tomoda
authors.affiliation: Department of Infectious Disease, Division of Microbiology1, First Department of Internal Medicine2, Laboratory of Electron Microscopy4 and Department of Anatomy5, Kyorin University School of Medicine,Tokyo 181-8611, Japan 3Division of Molecular Microbiology, Department of Basic Laboratory Sciences, Graduate School of Medicine, Osaka University, 1-7 Yamadaoka, Suita, Osaka 565-0871, Japan 6Department of Virology I, National Institute of Infectious Diseases, Tokyo 162-8642, Japan 7Department of Biochemistry, Tokyo Medical University, Tokyo 160-0022, Japan.
authors.forename: Shigeru
authors.initials: S
authors.lastname: Kamiya
mesh_terms: D047072:Aromatase Inhibitors / Q000494:pharmacology; D002460:Cell Line; D016993:Chlamydophila pneumoniae / Q000187:drug effects / Q000254:growth & development* / Q000648:ultrastructure; D004847:Epithelial Cells / Q000382:microbiology; D006801:Humans; D008854:Microscopy, Electron; D009000:Monocytes / Q000382:microbiology; D010078:Oxazines / Q000494:pharmacology*; D012329:RNA, Bacterial / Q000235:genetics; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D014364:Tryptophan / Q000494:pharmacology
chemical_list: D047072:Aromatase Inhibitors; D010078:Oxazines; D012329:RNA, Bacterial; D012336:RNA, Ribosomal, 16S; C005512:3-aminophenoxazone; D014364:Tryptophan