title: Mice Survival and Plasmatic Cytokine Secretion in a "Two Hit" Model of Sepsis Depend on Intratracheal Pseudomonas Aeruginosa Bacterial Load.
abstract: Sepsis is characterized by pro- and anti-inflammatory responses following infection. While inflammation is responsible for widespread organ damage, anti-inflammatory mediators lead to immunoparalysis increasing susceptibility to secondary infections (nosocomial pneumonia). We aimed to investigate the impact of bacterial load on survival and cytokine release in a two-hit murine (C57BL/6J) model of CLP followed by P. aeruginosa pneumonia. Plasmatic TNFα, IL-6, IL-10, sTNFr I and II were quantified until 13 days. At D5, splenocytes were processed for immunological assays or mice were intratracheally instilled with Pseudomonas aeruginosa (5.106, 2.107 and 108 CFU) to evaluate survival and cytokines production. TNFα, sTNFrs, IL-6 and IL-10 increased 2h post CLP. TNFα and sTNFrs declined respectively one and two days later. In CLP mice, IL-6 and IL-10 remained high for the whole experiment, as compared to Sham. At D5, for CLP mice, whereas total T cells population (CD3+) decreased, Treg fraction (CD4+/CD25+) increased. In parallel, T cells proliferation and LPS-stimulated splenocytes ability to release TNFα decreased. At D13, survival was 100% after 5.106 CFU, 50% for CLP mice after 2.107 CFU and 0% for CLP and Sham after 108 CFU. After instillation, IL-10 and IL-6 increased and appeared to be dose and time dependent. Pseudomonas was detected in all CLP and Sham's lungs; in spleen and liver only in CLP at 2.107 CFU, and in CLP and Sham at 108 CFU. We demonstrated that post-CLP immunosuppression followed by Pseudomonas aeruginosa lung instillation increases mortality reactivates cytokines secretion and is associated with systemic dissemination in septic mice depending on bacterial load.
authors: Université de Lyon, APCSe, Pulmonary and Cardiovascular Agression in Sepsis, VetAgro Sup-Campus Vétérinaire de Lyon, Marcy l'Étoile, France.
authors: Damien
authors: D
authors: Restagno
authors: Hospices Civils de Lyon, Immunology Laboratory, Hôpital Edouard Herriot, Lyon, France.
authors: Fabienne
authors: F
authors: Venet
authors: Université de Lyon, APCSe, Pulmonary and Cardiovascular Agression in Sepsis, VetAgro Sup-Campus Vétérinaire de Lyon, Marcy l'Étoile, France.
authors: Christian
authors: C
authors: Paquet
authors: Université de Lyon, APCSe, Pulmonary and Cardiovascular Agression in Sepsis, VetAgro Sup-Campus Vétérinaire de Lyon, Marcy l'Étoile, France.
authors: Ludovic
authors: L
authors: Freyburger
authors: Université de Lyon, APCSe, Pulmonary and Cardiovascular Agression in Sepsis, VetAgro Sup-Campus Vétérinaire de Lyon, Marcy l'Étoile, France.|Hospices Civils de Lyon, Service d'Anesthésie-Réanimation, Hôpital Edouard Herriot, Lyon, France.
authors: Bernard
authors: B
authors: Allaouchiche
authors: Hospices Civils de Lyon, Immunology Laboratory, Hôpital Edouard Herriot, Lyon, France.
authors: Guillaume
authors: G
authors: Monneret
authors: Université de Lyon, APCSe, Pulmonary and Cardiovascular Agression in Sepsis, VetAgro Sup-Campus Vétérinaire de Lyon, Marcy l'Étoile, France.
authors: Jeanne-Marie
authors: JM
authors: Bonnet
authors: Université de Lyon, APCSe, Pulmonary and Cardiovascular Agression in Sepsis, VetAgro Sup-Campus Vétérinaire de Lyon, Marcy l'Étoile, France.
authors: Vanessa
authors: V
authors: Louzier
mesh_terms: D000818:Animals; D058491:Bacterial Load; D049109:Cell Proliferation; D002478:Cells, Cultured; D016207:Cytokines / Q000097:blood*; D004195:Disease Models, Animal; D051379:Mice; D008810:Mice, Inbred C57BL; D011552:Pseudomonas Infections / Q000276:immunology*; D011550:Pseudomonas aeruginosa / Q000254:growth & development*; D018805:Sepsis / Q000150:complications* / Q000276:immunology; D013154:Spleen / Q000166:cytology / Q000276:immunology; D016019:Survival Analysis; D013601:T-Lymphocytes / Q000166:cytology / Q000276:immunology
chemical_list: D016207:Cytokines