Molecular Imaging with Kupffer Cell-Targeting Nanobodies for Diagnosis and Prognosis in Mouse Models of Liver Pathogenesis

• Published in: Molecular imaging and biology Vol. 19; no. 1; pp. 49 - 58
• Main Authors: Zheng, Fang, Sparkes, Amanda, De Baetselier, Patrick, Schoonooghe, Steve, Stijlemans, Benoit, Muyldermans, Serge, Flamand, Véronique, Van Ginderachter, Jo A, Devoogdt, Nick, Raes, Geert, Beschin, Alain
• Format: Journal Article
• Language: English
• Published: New York Springer US 01-02-2017; Springer Nature B.V
• Subjects: Animals; CD11b Antigen - metabolism; Cell Count; Choline; Concanavalin A; Disease Models, Animal; Hepatitis, Autoimmune - pathology; Imaging; Kupffer Cells - metabolism; Lectins, C-Type - metabolism; Liver - metabolism; Liver - pathology; Male; Medicine; Medicine & Public Health; Methionine - deficiency; Mice, Inbred C57BL; Molecular Imaging - methods; Prognosis; Radiology; Research Article; Single-Domain Antibodies - chemistry; Tissue Distribution; V-set and immunoglobulin domain-containing 4 (Vsig4); C-type lectin domain family, 4 member F (Clec4F); Hepatitis; Nanobodies (Nbs); Steatohepatitis; Methionine choline deficiency (MCD); Concanavalin A
• ISSN: 1536-1632; 1860-2002
• DOI: 10.1007/s11307-016-0976-3

Purpose
Kupffer cells (KCs), the liver resident macrophages, are important mediators of tissue homeostasis and pathogen clearance. However, depending on the inflammatory stimuli, KCs have been involved in divergent hepato-protective or hepato-destructive immune responses. The versatility of KCs in response to environmental triggers, in combination with the specific biomarkers they express, make these macrophages attractive
in vivo
targets for non-invasive monitoring of liver inflammation or pathogenicity. This study aims to determine whether V-set and Ig domain-containing 4 (Vsig4) and C-type lectin domain family (Clec) 4, member F (Clec4F) can be used as imaging biomarkers for non-invasive monitoring of KCs during distinct liver inflammation models.
Procedure
Flow cytometry (FACS), immuno-histochemistry (IHC), and single-photon emission computed tomography (SPECT) with Tc-99m labeled anti-Vsig4 or anti-Clec4F nanobodies (Nbs) was performed to evaluate in mice KC dynamics in concanavalin A (ConA)-induced hepatitis and in non-alcoholic steatohepatitis induced
via
methionine choline deficiency (MCD).
Results
In homeostatic mice, Nbs targeting Clec4F were found to accumulate and co-localize with Vsig4-targeting Nbs only in the liver. Upon induction of acute hepatitis using ConA, down-regulation of the
in vivo
Nb imaging signal was observed, reflecting reduction in KC numbers as confirmed by FACS and IHC. On the other hand, induction of steatohepatitis resulted in higher signals in the liver corresponding to higher density of KCs. The Nb-imaging signals returned to normal levels after resolution of the investigated liver diseases.
Conclusions
Anti-Clec4F and anti-Vsig4 Nbs targeting KCs as molecular imaging biomarkers could allow non-invasive monitoring/staging of liver pathogenesis.