title: Ex vivo lung function measurements in precision-cut lung slices (PCLS) from chemical allergen-sensitized mice represent a suitable alternative to in vivo studies.
abstract: A wide range of industrial chemicals can induce respiratory allergic reactions. Hence, there is an urgent need for methods identifying and characterizing the biological action of chemicals in the lung. Here, we present an easy, reliable alternative method to measure lung function changes ex vivo after exposure to chemical allergens and compare this to invasive in vivo measurements after sensitization with the industrial chemicals trimellitic anhydride (TMA) and 2,4-dinitrochlorobenzene (DNCB). Female BALB/c mice were sensitized epicutaneously with the respiratory allergen TMA and the contact sensitizer DNCB. The early allergic response to TMA and DNCB was registered in vivo and ex vivo on day 21 after inhalational challenge with dry standardized aerosols or after exposure of precision-cut lung slices (PCLS) to dissolved allergen. Airway hyperresponsiveness (AHR) to increasing doses of methacholine (MCh) was measured on the next day in vivo and ex vivo. Bronchoalveolar lavage (BAL) was performed for immunological characterization of local inflammation. TMA-sensitized mice showed AHR to MCh in vivo (ED(50): 0.06 microg MCh vs. 0.21 microg MCh in controls) and in PCLS (EC(50): 0.24 microM MCh vs. 0.4 microM MCh). TMA-treated animals showed increased numbers of eosinophils (12.8 x 10(4) vs. 0.7 x 10(4)) and elevated eotaxin-2 concentrations (994 pg/ml vs. 167 pg/ml) in BAL fluid 24 h after allergen challenge. In contrast, none of these parameters differed after sensitization with DNCB. The present study suggests that the effects of low molecular weight allergens, like TMA and DNCB, on ex vivo lung functions tested in PCLS reflect the in vivo situation.
authors.affiliation: Department of Immunology, Allergology and Immunotoxicology, Fraunhofer Institute of Toxicology and Experimental Medicine, 30625 Hannover, Germany.
authors.forename: M
authors.initials: M
authors.lastname: Henjakovic
authors.forename: C
authors.initials: C
authors.lastname: Martin
authors.forename: H G
authors.initials: HG
authors.lastname: Hoymann
authors.forename: K
authors.initials: K
authors.lastname: Sewald
authors.forename: A R
authors.initials: AR
authors.lastname: Ressmeyer
authors.forename: C
authors.initials: C
authors.lastname: Dassow
authors.forename: G
authors.initials: G
authors.lastname: Pohlmann
authors.forename: N
authors.initials: N
authors.lastname: Krug
authors.forename: S
authors.initials: S
authors.lastname: Uhlig
authors.forename: A
authors.initials: A
authors.lastname: Braun
mesh_terms: D000485:Allergens / Q000633:toxicity*; D000818:Animals; D016535:Bronchial Hyperreactivity; D001992:Bronchoalveolar Lavage Fluid; D016084:Bronchoconstriction / Q000187:drug effects; D004137:Dinitrochlorobenzene / Q000633:toxicity*; D004305:Dose-Response Relationship, Drug; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D066298:In Vitro Techniques; D008168:Lung / Q000187:drug effects* / Q000502:physiology; D016210:Methacholine Chloride / Q000633:toxicity*; D051379:Mice; D008807:Mice, Inbred BALB C; D010796:Phthalic Anhydrides / Q000633:toxicity*; D011014:Pneumonia / Q000139:chemically induced; D012129:Respiratory Function Tests
chemical_list: D000485:Allergens; D004137:Dinitrochlorobenzene; D010796:Phthalic Anhydrides; D016210:Methacholine Chloride; C015559:trimellitic anhydride