Abstract
Cell-to-cell interactions between the immune and nervous systems are increasingly recognized for their importance in health and disease. Assessment of cellular neuro-immune interactions can be aided by co-culture of two (or more) cells in an in vitro model system that preserves the morphology of neuronal cells. Here we describe methods to investigate the cytotoxic effector functions of natural killer cells on sensory neurons isolated from syngeneic embryonic and adult mice. We present methods for the morphological analysis of axon fragmentation (pruning) and dynamic cell function via live confocal calcium imaging. These techniques can easily be adapted to study interactions between other combinations of immune cell subsets and neuronal populations.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Davies AJ, Rinaldi S, Costigan M, Oh SB (2020) Cytotoxic immunity in peripheral nerve injury and pain. Front Neurosci 14:142
Poli A, Kmiecik J, Domingues O, Hentges F, Blery M, Chekenya M, Boucraut J, Zimmer J (2013) NK cells in central nervous system disorders. J Immunol 190(11):5355–5362
** WN, Shi K, He W, Sun JH, Van Kaer L, Shi FD, Liu Q (2021) Neuroblast senescence in the aged brain augments natural killer cell cytotoxicity leading to impaired neurogenesis and cognition. Nat Neurosci 24(1):61–73
Garofalo S, Cocozza G, Porzia A, Inghilleri M, Raspa M, Scavizzi F, Aronica E, Bernardini G, Peng L, Ransohoff RM, Santoni A, Limatola C (2020) Natural killer cells modulate motor neuron-immune cell cross talk in models of amyotrophic lateral sclerosis. Nat Commun 11(1):1773
Earls RH, Menees KB, Chung J, Gutekunst CA, Lee HJ, Hazim MG, Rada B, Wood LB, Lee JK (2020) NK cells clear alpha-synuclein and the depletion of NK cells exacerbates synuclein pathology in a mouse model of alpha-synucleinopathy. Proc Natl Acad Sci U S A 117(3):1762–1771
Gross CC, Schulte-Mecklenbeck A, Runzi A, Kuhlmann T, Posevitz-Fejfar A, Schwab N, Schneider-Hohendorf T, Herich S, Held K, Konjevic M, Hartwig M, Dornmair K, Hohlfeld R, Ziemssen T, Klotz L, Meuth SG, Wiendl H (2016) Impaired NK-mediated regulation of T-cell activity in multiple sclerosis is reconstituted by IL-2 receptor modulation. Proc Natl Acad Sci U S A 113(21):E2973–E2982
Davies AJ, Kim HW, Gonzalez-Cano R, Choi J, Back SK, Roh SE, Johnson E, Gabriac M, Kim MS, Lee J, Lee JE, Kim YS, Bae YC, Kim SJ, Lee KM, Na HS, Riva P, Latremoliere A, Rinaldi S, Ugolini S, Costigan M, Oh SB (2019) Natural killer cells degenerate intact sensory afferents following nerve injury. Cell 176(4):716–728 e18
Schneider CA, Rasband WS, Eliceiri KW (2012) NIH image to ImageJ: 25 years of image analysis. Nat Methods 9(7):671–675
The AVMA Guidelines for the Euthanasia of Animals: 2020 Edition. American Veterinary Medical Association Available from: https://www.avma.org/sites/default/files/2020-02/Guidelines-on-Euthanasia-2020.pdf. Accessed: 08/04/2021
Acknowledgements
The authors would like to thank Dr. Steven West for assistance with the macro script.
This work was supported by the National Research Foundation of Korea grants (NRF-2018R1A5A2024418 and NRF-2021R1A2C3003334) funded by the Korean government MSIT (Ministry of Science and ICT). A.J.D receives support from the National Institute for Health Research (NIHR) Oxford Biomedical Research Centre (BRC) and Medical Research Council United Kingdom and the UKRI Future Leaders Fellowship programme (Grant number MR/V02552X/1).
Author information
Authors and Affiliations
Corresponding authors
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2022 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Kim, H.W., Davies, A.J., Oh, S.B. (2022). In Vitro Visualization of Cell-to-Cell Interactions Between Natural Killer Cells and Sensory Neurons. In: Shimasaki, N. (eds) Natural Killer (NK) Cells. Methods in Molecular Biology, vol 2463. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2160-8_18
Download citation
DOI: https://doi.org/10.1007/978-1-0716-2160-8_18
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-2159-2
Online ISBN: 978-1-0716-2160-8
eBook Packages: Springer Protocols