Ex vivo testing of immune responses in precision-cut lung slices

Journal Article · Fri Aug 15 00:00:00 EDT 2008 · Toxicology and Applied Pharmacology

DOI:https://doi.org/10.1016/j.taap.2008.04.003· OSTI ID:21140928

Henjakovic, M; Sewald, K; Switalla, S; Kaiser, D; Mueller, M; Veres, T Z ^[1]; Martin, C; Uhlig, S ^[2]; Krug, N ^[1]

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1. Fraunhofer Institute of Toxicology and Experimental Medicine, Department of Immunology, Allergology and Immunotoxicology, Nikolai-Fuchs-Str. 1, 30625 Hannover (Germany)
2. Institute of Pharmacology and Toxicology, RWTH Aachen, Wendlingweg 2, 52074 Aachen (Germany)

The aim of this study was the establishment of precision-cut lung slices (PCLS) as a suitable ex vivo alternative approach to animal experiments for investigation of immunomodulatory effects. For this purpose we characterized the changes of cytokine production and the expression of cell surface markers after incubation of PCLS with immunoactive substances lipopolysaccharide (LPS), macrophage-activating lipopeptide-2 (MALP-2), interferon {gamma} (IFN{gamma}), and dexamethasone. Viability of PCLS from wild-type and CD11c-enhanced yellow fluorescent protein (CD11-EYFP)-transgenic mice was controlled by measurement of lactate dehydrogenase (LDH) enzyme activity and live/dead fluorescence staining using confocal microscopy. Cytokines and chemokines were detected with Luminex technology and ELISA. Antigen presenting cell (APC) markers were investigated in living mouse PCLS in situ using confocal microscopy. LPS triggered profound pro-inflammatory effects in PCLS. Dexamethasone prevented LPS-induced production of cytokines/chemokines such as interleukin (IL)-5, IL-1{alpha}, TNF{alpha}, IL-12(p40), and RANTES in PCLS. Surface expression of MHC class II, CD40, and CD11c, but not CD86 was present in APCs of naive PCLS. Incubation with LPS enhanced specifically the expression of MHC class II on diverse cells. MALP-2 only failed to alter cytokine or chemokine levels, but was highly effective in combination with IFN{gamma} resulting in increased levels of TNF{alpha}, IL-12(p40), RANTES, and IL-1{alpha}. PCLS showed characteristic responses to typical pro-inflammatory stimuli and may thus provide a suitable ex vivo technique to predict the immunomodulatory potency of inhaled substances.

• OSTI ID: 21140928
• Journal Information: Toxicology and Applied Pharmacology, Vol. 231, Issue 1; Other Information: DOI: 10.1016/j.taap.2008.04.003; PII: S0041-008X(08)00160-9; Copyright (c) 2008 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved; Country of input: International Atomic Energy Agency (IAEA); ISSN 0041-008X
• Country of Publication: United States
• Language: English

Related Subjects

60 APPLIED LIFE SCIENCES
ACCURACY
ANTIGENS
DEXAMETHASONE
ENZYME ACTIVITY
ENZYME IMMUNOASSAY
FLUORESCENCE
INCUBATION
INFLAMMATION
INTERFERON
LACTATE DEHYDROGENASE
LUNGS
MACROPHAGES
MICROSCOPY
TRANSGENIC MICE