Abstract
Introduction
Immune cells utilize acetylcholine as a paracrine-signaling molecule. Many white blood cells express components of the cholinergic signaling pathway, and these are up-regulated when immune cells are activated. However, in vivo molecular imaging of cholinergic signaling in the context of inflammation has not previously been investigated.
Methods
We performed positron emission tomography (PET) using the glucose analogue 18F-FDG, and 11C-donepezil and 18F-FEOBV, markers of acetylcholinesterase and the vesicular acetylcholine transporter, respectively. Mice were inoculated subcutaneously with Staphylococcus aureus, and PET scanned at 24, 72, 120, and 144 h post-inoculation. Four pigs with post-operative abscesses were also imaged. Finally, we present initial data from human patients with infections, inflammation, and renal and lung cancer.
Results
In mice, the FDG uptake in abscesses peaked at 24 h and remained stable. The 11C-donepezil and 18F-FEOBV uptake displayed progressive increase, and at 120–144 h was nearly at the FDG level. Moderate 11C-donepezil and slightly lower 18F-FEOBV uptake were seen in pig abscesses. PCR analyses suggested that the 11C-donepezil signal in inflammatory cells is derived from both acetylcholinesterase and sigma-1 receptors. In humans, very high 11C-donepezil uptake was seen in a lobar pneumonia and in peri-tumoral inflammation surrounding a non-small cell lung carcinoma, markedly superseding the 18F-FDG uptake in the inflammation. In a renal clear cell carcinoma no 11C-donepezil uptake was seen.
Discussion
The time course of cholinergic tracer accumulation in murine abscesses was considerably different from 18F-FDG, demonstrating in the 11C-donepezil and 18F-FEOBV image distinct aspects of immune modulation. Preliminary data in humans strongly suggest that 11C-donepezil can exhibit more intense accumulation than 18F-FDG at sites of chronic inflammation. Cholinergic PET imaging may therefore have potential applications for basic research into cholinergic mechanisms of immune modulation, but also clinical applications for diagnosing infections, inflammatory disorders, and cancer inflammation.
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Abbreviations
- AChE:
-
Acetylcholinesterase
- CFU:
-
Colony forming unit
- MRI:
-
Magnetic Resonance Imaging
- PET:
-
Positron Emission Tomography
- PMN:
-
Polymorph-nuclear cells
- ROI:
-
2D region of interest
- SPECT:
-
Single photon emission computed tomography
- SUV:
-
Standardized uptake value
- VAChT:
-
Vesicular acetylcholine transporter
- VOI:
-
3D volume of interest
- WBC:
-
White blood cells
References
Sobic-Saranovic DP, Grozdic IT, Videnovic-Ivanov J, Vucinic-Mihailovic V, Artiko VM, Saranovic DZ, et al. Responsiveness of FDG PET/CT to treatment of patients with active chronic sarcoidosis. Clin Nucl Med. 2013;38:516–21. doi:10.1097/RLU.0b013e31828731f5.
Bleeker-Rovers CP, Vos FJ, Mudde AH, Dofferhoff AS, de Geus-Oei LF, Rijnders AJ, et al. A prospective multi-centre study of the value of FDG-PET as part of a structured diagnostic protocol in patients with fever of unknown origin. Eur J Nucl Med Mol Imaging. 2007;34:694–703. doi:10.1007/s00259-006-0295-z.
Lehmann P, Buchtala S, Achajew N, Haerle P, Ehrenstein B, Lighvani H, et al. 18F-FDG PET as a diagnostic procedure in large vessel vasculitis-a controlled, blinded re-examination of routine PET scans. Clin Rheumatol. 2011;30:37–42. doi:10.1007/s10067-010-1598-9.
Jamar F, Buscombe J, Chiti A, Christian PE, Delbeke D, Donohoe KJ, et al. EANM/SNMMI guideline for 18F-FDG use in inflammation and infection. J Nucl Med. 2013;54:647–58. doi:10.2967/jnumed.112.112524.
Bensinger SJ, Christofk HR. New aspects of the Warburg effect in cancer cell biology. Semin Cell Dev Biol. 2012;23:352–61. doi:10.1016/j.semcdb.2012.02.003.
Kawashima K, Fujii T, Moriwaki Y, Misawa H. Critical roles of acetylcholine and the muscarinic and nicotinic acetylcholine receptors in the regulation of immune function. Life Sci. 2012;91:1027–32. doi:10.1016/j.lfs.2012.05.006.
Kawashima K, Fujii T. Expression of non-neuronal acetylcholine in lymphocytes and its contribution to the regulation of immune function. Front Biosci. 2004;9:2063–85.
Szelenyi JG, Bartha E, Hollan SR. Acetylcholinesterase activity of lymphocytes: an enzyme characteristic of T-cells. Br J Haematol. 1982;50:241–5.
Tayebati SK, El-Assouad D, Ricci A, Amenta F. Immunochemical and immunocytochemical characterization of cholinergic markers in human peripheral blood lymphocytes. J Neuroimmunol. 2002;132:147–55.
Fujii TWY, Fujimoto K, Kawashima K. Expression of acetylcholine in lymphocytes and modulation of an independent lymphocytic cholinergic activity by immunological stimulation. Biog Amine. 2003;17:373–86.
Mori T, Maeda J, Shimada H, Higuchi M, Shinotoh H, Ueno S, et al. Molecular imaging of dementia. Psychogeriatrics Off J Jpn Psychogeriatr Soc. 2012;12:106–14. doi:10.1111/j.1479-8301.2012.00409.x.
Gjerloff T, Jakobsen S, Nahimi A, Munk OL, Bender D, Alstrup AK, et al. In vivo imaging of human acetylcholinesterase density in peripheral organs using 11C-donepezil: dosimetry, biodistribution, and kinetic analyses. J Nucl Med. 2014;55:1818–24. doi:10.2967/jnumed.114.143859.
Gjerloff T, Fedorova T, Knudsen K, Munk OL, Nahimi A, Jacobsen S, et al. Imaging acetylcholinesterase density in peripheral organs in Parkinson’s disease with 11C-donepezil PET. Brain. 2015;138:653–63. doi:10.1093/brain/awu369.
Petrou M, Frey KA, Kilbourn MR, Scott PJ, Raffel DM, Bohnen NI, et al. In vivo imaging of human cholinergic nerve terminals with (−)-5-(18)F-fluoroethoxybenzovesamicol: biodistribution, dosimetry, and tracer kinetic analyses. J Nucl Med. 2014;55:396–404. doi:10.2967/jnumed.113.124792.
Pellegrino D, Bonab AA, Dragotakes SC, Pitman JT, Mariani G, Carter EA. Inflammation and infection: imaging properties of 18F-FDG-labeled white blood cells versus 18F-FDG. J Nucl Med. 2005;46:1522–30.
Nippe N, Varga G, Holzinger D, Loffler B, Medina E, Becker K, et al. Subcutaneous infection with S. aureus in mice reveals association of resistance with influx of neutrophils and Th2 response. J Invest Dermatol. 2011;131:125–32. doi:10.1038/jid.2010.282.
Molne L, Verdrengh M, Tarkowski A. Role of neutrophil leukocytes in cutaneous infection caused by Staphylococcus aureus. Infect Immun. 2000;68:6162–7.
McLoughlin RM, Solinga RM, Rich J, Zaleski KJ, Cocchiaro JL, Risley A, et al. CD4+ T cells and CXC chemokines modulate the pathogenesis of Staphylococcus aureus wound infections. Proc Natl Acad Sci U S A. 2006;103:10408–13. doi:10.1073/pnas.0508961103.
Archer NK, Harro JM, Shirtliff ME. Clearance of Staphylococcus aureus nasal carriage is T cell dependent and mediated through interleukin-17A expression and neutrophil influx. Infect Immun. 2013;81:2070–5. doi:10.1128/IAI.00084-13.
Fujii T, Yamada S, Watanabe Y, Misawa H, Tajima S, Fujimoto K, et al. Induction of choline acetyltransferase mRNA in human mononuclear leukocytes stimulated by phytohemagglutinin, a T-cell activator. J Neuroimmunol. 1998;82:101–7.
Suga K, Kawakami Y, Hiyama A, Sugi K, Okabe K, Matsumoto T, et al. Dual-time point 18F-FDG PET/CT scan for differentiation between 18F-FDG-avid non-small cell lung cancer and benign lesions. Ann Nucl Med. 2009;23:427–35. doi:10.1007/s12149-009-0260-6.
Nguyen NC, Kaushik A, Wolverson MK, Osman MM. Is there a common SUV threshold in oncological FDG PET/CT, at least for some common indications? A retrospective study. Acta Oncol. 2011;50:670–7. doi:10.3109/0284186X.2010.550933.
Tateishi U, Hasegawa T, Seki K, Terauchi T, Moriyama N, Arai Y. Disease activity and 18F-FDG uptake in organising pneumonia: semi-quantitative evaluation using computed tomography and positron emission tomography. Eur J Nucl Med Mol Imaging. 2006;33:906–12. doi:10.1007/s00259-006-0073-y.
Bremnes RM, Al-Shibli K, Donnem T, Sirera R, Al-Saad S, Andersen S, et al. The role of tumor-infiltrating immune cells and chronic inflammation at the tumor site on cancer development, progression, and prognosis: emphasis on non-small cell lung cancer. J Thoracic Oncol Off Publ Int Assoc Stud Lung Cancer. 2011;6:824–33. doi:10.1097/JTO.0b013e3182037b76.
Munoz-Delgado E, Montenegro MF, Campoy FJ, Moral-Naranjo MT, Cabezas-Herrera J, Kovacs G, et al. Expression of cholinesterases in human kidney and its variation in renal cell carcinoma types. FEBS J. 2010;277:4519–29. doi:10.1111/j.1742-4658.2010.07861.x.
Hiraoka K, Okamura N, Funaki Y, Watanuki S, Tashiro M, Kato M, et al. Quantitative analysis of donepezil binding to acetylcholinesterase using positron emission tomography and [5-(11)C-methoxy]donepezil. Neuroimage. 2009;46:616–23. doi:10.1016/j.neuroimage.2009.03.006.
Ishikawa M, Sakata M, Ishii K, Kimura Y, Oda K, Toyohara J, et al. High occupancy of sigma1 receptors in the human brain after single oral administration of donepezil: a positron emission tomography study using [11C]SA4503. Int J Neuropsychopharmacol / Off Sci J Coll Int Neuropsychopharmacol (CINP). 2009;12:1127–31. doi:10.1017/S1461145709990204.
Carayon P, Bouaboula M, Loubet JF, Bourrie B, Petitpretre G, Le Fur G, et al. The sigma ligand SR 31747 prevents the development of acute graft-versus-host disease in mice by blocking IFN-gamma and GM-CSF mRNA expression. Int J Immunopharmacol. 1995;17:753–61.
Zhu LX, Sharma S, Gardner B, Escuadro B, Atianzar K, Tashkin DP, et al. IL-10 mediates sigma 1 receptor-dependent suppression of antitumor immunity. J Immunol. 2003;170:3585–91.
Toyohara J, Elsinga PH, Ishiwata K, Sijbesma JW, Dierckx RA, van Waarde A. Evaluation of 4′-[methyl-11C]thiothymidine in a rodent tumor and inflammation model. J Nucl Med. 2012;53:488–94. doi:10.2967/jnumed.111.098426.
Acknowledgments
We acknowledge laboratory technician Mette Simonsen for invaluable help on PET/MRI imaging.
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The study was funded by the Danish Medical Research Council and the Lundbeck foundation. Per Borghammer has served as a consultant for F. Hoffmann – La Roche Ltd. All other co-authors had no conflicts of interest concerning the present study. Permission for the studies of mice and pigs were obtained from the Danish Animal Experiments Inspectorate (j. nr. 2013-15-2934-00878). All applicable international, national, and/or institutional guidelines for the care and use of animals were followed. All human studies were approved by the Central Denmark Region Committee on Health Research and the Danish Health and Medicines Authority, and followed the ethical standards of the institutional and national research committee and the 1964 Helsinki declaration and its later amendments. Written informed consent was obtained from all participants prior to enrollment.
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Jørgensen, N.P., Alstrup, A.K.O., Mortensen, F.V. et al. Cholinergic PET imaging in infections and inflammation using 11C-donepezil and 18F-FEOBV. Eur J Nucl Med Mol Imaging 44, 449–458 (2017). https://doi.org/10.1007/s00259-016-3555-6
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DOI: https://doi.org/10.1007/s00259-016-3555-6