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

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Abstract

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.

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Acknowledgments

The authors thank Cindy Peleman, Ella Omasta, Marie-Therese Detobel, Maria Slazak, Victor Orimoloye, Lea Brys, Yvon Elkrim and Nadia Abou for technical and secretarial assistance, Dr. Marie-Aline Laute (ULB) for GOT/GPT measurements and Ir. Chloé Abels for providing Clec4F-DTR and littermate mice. Support grants were received from the Agency for Innovation by Science and Technology, FWO-Vlaanderen, the Interuniversity Attraction Poles and the Strategic Research Program (SRP3). FZ received a scholarship from the Top-Notch Student Scholarship Fund of China Scholarship Council.

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Authors and Affiliations

  1. Laboratory of Cellular and Molecular Immunology, Vrije Universiteit Brussel, Pleinlaan 2, Building E, 8th floor, Brussels, Belgium

    Fang Zheng, Amanda Sparkes, Patrick De Baetselier, Steve Schoonooghe, Benoit Stijlemans, Serge Muyldermans, Jo A Van Ginderachter, Geert Raes & Alain Beschin

  2. Myeloid Cell Immunology Laboratory, VIB Inflammation Research Center, Ghent, Belgium

    Fang Zheng, Amanda Sparkes, Patrick De Baetselier, Steve Schoonooghe, Benoit Stijlemans, Jo A Van Ginderachter, Geert Raes & Alain Beschin

  3. Institute of Medical Immunology, ULB, Gosselies, Belgium

    Véronique Flamand

  4. In Vivo Cellular and Molecular Imaging Center, VUB, Brussels, Belgium

    Nick Devoogdt

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  1. Fang Zheng
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  2. Amanda Sparkes
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Corresponding author

Correspondence to Alain Beschin.

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Conflict of Interest

The authors declare that they have no conflict of interest.

Human and Animal Rights and Informed Consent

All applicable institutional and/or national guidelines for the care and use of animals were followed (ECPVA guidelines (CETS no. 123) and approved by the VUB Ethical Committee (Lab Accreditation number: LA1210220).

Additional information

Fang Zheng and Amanda Sparkes share first authorship

Nick Devoogdt, Geert Raes and Alain Beschin share senior authorship

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Zheng, F., Sparkes, A., De Baetselier, P. et al. Molecular Imaging with Kupffer Cell-Targeting Nanobodies for Diagnosis and Prognosis in Mouse Models of Liver Pathogenesis. Mol Imaging Biol 19, 49–58 (2017). https://doi.org/10.1007/s11307-016-0976-3

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  • DOI: https://doi.org/10.1007/s11307-016-0976-3

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