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Abstract

Immunology and Infection

NAD(P)H Fluorescence Lifetime Imaging for the Metabolic Analysis of the Murine Intestine and Parasites During Nematode Infection

Published: September 1st, 2023

DOI:

10.3791/64982

1Biophysical Analytics, Deutsches Rheuma-Forschungszentrum, Berlin - A Leibniz Institute, 2Dynamic and Functional In Vivo Imaging, Department of Veterinary Medicine, Institute for Veterinary Physiology, Freie Universität Berlin, 3Institute of Immunology, Department of Veterinary Medicine, Freie Universität Berlin, 4Laboratory for Immune Dynamics, Deutsches Rheuma-Forschungszentrum, Berlin - A Leibniz Institute, 5Intravital Microscopy and Immune Dynamics, Department of Rheumatology and Clinical Immunology, Charité - Universitätsmedizin Berlin, corporate member of Freie Universität Berlin and Humboldt Universität zu Berlin

Abstract

Parasites generally have a negative effect on the health of their host. They represent a huge health burden, as they globally affect the health of the infested human or animal in the long term and, thus, impact agricultural and socio-economic outcomes. However, parasite-driven immune-regulatory effects have been described, with potential therapeutic relevance for autoimmune diseases. While the metabolism in both the host and parasites contributes to their defense and is the basis for nematode survival in the intestine, it has remained largely understudied due to a lack of adequate technologies. We have developed and applied NAD(P)H fluorescence lifetime imaging to explanted murine intestinal tissue during infection with the natural nematode Heligmosomoides polygyrus to study the metabolic processes in both the host and parasites in a spatially resolved manner. The exploitation of the fluorescence lifetime of the co-enzymes nicotinamide adenine dinucleotide (NADH) and nicotinamide adenine dinucleotide phosphate (NADPH), hereafter NAD(P)H, which are preserved across species, depends on their binding status and the binding site on the enzymes catalyzing metabolic processes. Focusing on the most abundantly expressed NAD(P)H-dependent enzymes, the metabolic pathways associated with anaerobic glycolysis, oxidative phosphorylation/aerobic glycolysis, and NOX-based oxidative burst, as a major defense mechanism, were distinguished, and the metabolic crosstalk between the host and parasite during infection was characterized.

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Keywords NAD P H Fluorescence Lifetime Imaging

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