title: Neutrophil gelatinase-associated lipocalin and interleukin-10 regulate intramacrophage Chlamydia pneumoniae replication by modulating intracellular iron homeostasis.
abstract: Neutrophil gelatinase-associated lipocalin (NGAL/Lipocalin-2/Lcn-2) is a 25kDa protein which is involved in host defence against certain Gram negative bacteria upon binding of iron loaded bacterial siderophores thereby limiting the availability of this essential nutrient to bacteria resulting in inhibition of their growth and pathogenicity. As iron is important for the growth of the intracellular bacterium Chlamydia pneumoniae we questioned whether Lcn-2 affects the course of this infection. We employed primary peritoneal macrophages obtained from wildtype and Lcn-2 -/- mice and RAW 264.7 cells which were infected with C. pneumoniae. In addition, we studied C. pneumoniae multiplication in vivo in mice receiving diets with varying iron contents. We analyzed C. pneumoniae numbers by immunohistochemistry and RT-PCR and studied the expression of iron metabolism and cytokine genes by RT-PCR, Western blot or ELISA. Infection with Chlamydiae ex vivo and in vivo revealed a significantly higher bacterial growth in peritoneal macrophages of Lcn-2 -/- than of wildtype mice. These differences were significantly more pronounced upon iron challenge, which stimulated bacterial growth. Accordingly, treatment with an anti-Lnc-2 antibody increased whereas addition of recombinant Lcn-2 reduced bacterial growth in infected macrophages. When investigating the underlying mechanisms we observed partly different expression of several iron metabolism genes between Lcn-2 +/+ and Lcn-2 -/- macrophages and most strikingly an increased formation of the anti-inflammatory cytokine IL-10 by Lcn-2 -/- macrophages. Upon treatment with an anti-IL10 antibody we experienced a significant increase of Chlamydial growth within Lcn-2 -/- macrophages along with a reduction of the major iron storage protein ferritin. Herein we provide first time evidence that Lcn-2 is involved in host defence against Chlamydia presumably by limiting the availability of iron to the pathogen. In the absence of Lcn-2, increased formation of IL-10 exerts protective effects by increasing the intracellular formation of ferritin, thereby reducing the access of iron for bacteria.
authors: Department of Internal Medicine VI, Clinical Immunology and Infectious Diseases, Medical University of Innsbruck, Austria.
authors: Rosa
authors: R
authors: Bellmann-Weiler
authors: Andrea
authors: A
authors: Schroll
authors: Sabine
authors: S
authors: Engl
authors: Manfred
authors: M
authors: Nairz
authors: Heribert
authors: H
authors: Talasz
authors: Markus
authors: M
authors: Seifert
authors: Guenter
authors: G
authors: Weiss
mesh_terms: D000209:Acute-Phase Proteins / Q000037:antagonists & inhibitors / Q000172:deficiency / Q000235:genetics / Q000276:immunology*; D000818:Animals; D000906:Antibodies / Q000494:pharmacology; D058491:Bacterial Load; D002460:Cell Line; D016993:Chlamydophila pneumoniae / Q000254:growth & development / Q000276:immunology / Q000378:metabolism*; D005260:Female; D005293:Ferritins / Q000235:genetics / Q000276:immunology*; D005786:Gene Expression Regulation / Q000187:drug effects; D006706:Homeostasis; D054884:Host-Pathogen Interactions; D016753:Interleukin-10 / Q000235:genetics / Q000276:immunology*; D007501:Iron / Q000378:metabolism* / Q000494:pharmacology; D019266:Iron, Dietary / Q000008:administration & dosage; D000071068:Lipocalin-2; D054834:Lipocalins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000276:immunology*; D017737:Macrophages, Peritoneal / Q000276:immunology* / Q000378:metabolism / Q000382:microbiology; D051379:Mice; D008810:Mice, Inbred C57BL; D015513:Oncogene Proteins / Q000037:antagonists & inhibitors / Q000172:deficiency / Q000235:genetics / Q000276:immunology*; D061251:Primary Cell Culture
chemical_list: D000209:Acute-Phase Proteins; D000906:Antibodies; C556062:IL10 protein, mouse; D019266:Iron, Dietary; D000071068:Lipocalin-2; D054834:Lipocalins; D015513:Oncogene Proteins; C071332:Lcn2 protein, mouse; D016753:Interleukin-10; D005293:Ferritins; D007501:Iron