Teasing Out the Interplay Between Natural Killer Cells and Nociceptor Neurons

Ali Ahmadi; Mohammad Balood; Katiane Roversi; Maryam Ahmadi; Moutih Rafei; Sebastien Talbot

doi:10.3791/63800

Teasing Out the Interplay Between Natural Killer Cells and Nociceptor Neurons

JoVE Journal
Neuroscience

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JoVE Journal Neuroscience

Teasing Out the Interplay Between Natural Killer Cells and Nociceptor Neurons

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09:40 min

June 30, 2022

DOI: 10.3791/63800-v

Ali Ahmadi¹, Mohammad Balood¹, Katiane Roversi¹, Maryam Ahmadi¹, Moutih Rafei¹, Sebastien Talbot¹

¹Department of Pharmacology and Physiology,Université de Montréal

Overview

This study explores the interaction between nociceptor neurons and NK cells in an inflammatory context using a co-culture approach. The aim is to understand how nociceptor neurons influence the anti-tumor functions of NK cells and the role of NK cells in eliminating injured neurons.

Key Study Components

Area of Science

Neuroscience
Cell Biology
Immunology

Background

Nociceptor neurons secrete neuropeptides that interact with NK cells.
Understanding the interplay between these cell types is crucial for insights into inflammation and tumor immunity.
The use of a co-culture system can simplify the investigation of these interactions.
Impacts on the functions of both cell types under different stimuli are of particular interest.

Purpose of Study

To devise a method for studying NK cell and nociceptor neuron interactions.
To explore how nociceptor neurons regulate NK cell activity, especially concerning tumor response.
To investigate NK cell effects on neuronal injury responses.

Methods Used

The study employs a co-culture system featuring nociceptor neurons and NK cells.
Cells are isolated and prepared from mouse spleen and dorsal root ganglion for analysis.
Key steps include cell isolation, culture preparations, and stimulation conditions.
Immunophenotyping is performed on NK cells using flow cytometry.

Main Results

The co-culture approach enables assessment of NK cell function modulation by nociceptor neurons.
Capsaicin treatment decreased NK cell expression of GM-CSF when co-cultured with DRG neurons.
Functional insights reveal interactions that could inform therapeutic approaches to inflammation and tumors.

Conclusions

This study demonstrates the utility of co-culture systems in elucidating cellular interactions in inflammation.
Insights from the study may enhance understanding of immune responses and neuronal interactions in health and disease.
The findings lay groundwork for further investigation into therapeutic targeting of these pathways.

Frequently Asked Questions

What are the advantages of the co-culture model?

The co-culture model allows for direct study of interactions between nociceptor neurons and NK cells in a controlled environment, facilitating insights into their functional dynamics.

How are nociceptor neurons prepared for co-culture?

Nociceptor neurons are isolated from dorsal root ganglia, processed with collagenase and dispase, and cultured in a laminin-coated plate for attachment.

What type of data is obtained from this study?

Data on NK cell functionality, specifically regarding their expression profiles and responses to nociceptor neuron signaling, are analyzed through flow cytometry.

Can this method be adapted for other cell types?

Yes, the co-culture methodology can be adapted to investigate interactions between various immune cells and neurons, broadening its applicability.

What are key limitations of this study?

Limitations could include the specific focus on mouse models, which may not completely replicate human conditions, and the complexity of in vivo interactions that might not fully translate in vitro.

How might findings from this study influence future research?

Results could pave the way for exploring therapeutic strategies aimed at enhancing NK cell responses to tumors or modulating pain through neuro-immune interactions.

Nociceptor neurons and NK cells actively interact in an inflammatory context. A co-culture approach enables studying this interplay.

Given that nociceptor neuron secrete various neuropeptide and that NK cell expressed the receptor for these neuropeptide, we decided to devise a co-culture method to study the interplay between the NK cell and the nociceptor neurons. Such a reductionist method could be useful to study how nociceptor neuron control the anti-tumor function of NK cell, and it could also be interesting to study how NK cell controlled the elimination of injured neurons. Demonstrating the procedure will be Ali Ahmadi, a former master's student from my lab.

To begin the natural killer or NK cell isolation and culture, collect the mouse spleen in a 1.5 milliliter microcentrifuge tube containing 500 microliters of sterile PBS, then homogenize the spleen using a pestle. Next, dilute the cells with one milliliter of sterile PBS and filter the mixture through a 50 micron cell strainer into a 50 milliliter conical tube. Top up the filtered homogenate volume to 10 milliliters using sterile PBS.

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