Investigating CD1d-dependent neuronal activation of natural killer T cells and its impact on neurodegenerative disease

Project Information

NKT-in-NDD

Grant agreement ID: 101202326

DOI

10.3030/101202326

Project terminated on 10 February 2026

EC signature date 27 February 2025

Start date 1 April 2025

End date 31 March 2027

Funded under

• Marie Skłodowska-Curie Actions (MSCA)

Total cost

No data

EU contribution

€ 247 553,28

Coordinated by

KOBENHAVNS UNIVERSITET
Denmark

Periodic Reporting for period 1 - NKT-in-NDD (Investigating CD1d-dependent neuronal activation of natural killer T cells and its impact on neurodegenerative disease)

Reporting period: 2025-04-01 to 2027-03-31

Summary of the context and overall objectives of the project

Neurodegenerative diseases (NDDs) are a broad category of neurological maladies, typically characterized by cognitive decline and motor deficits. NDDs affect over 55 million individuals worldwide, a number that is expected to triple over the following decades. Until recently, the brain was considered immune-privileged, but emerging scientific advancements in the field of neuroimmunology show that immune cells reside within and even infiltrate the central nervous system (CNS). Despite ever-growing evidence illuminating the critical interplay between the nervous and immune system in maintaining neural homeostasis, research efforts have been limited in this scope.

We recently identified that an immune-related gene, CD1d, is expressed by neurons. CD1d is recognized by natural killer T (NKT) cells, generally inducing an immunosuppressive response. We found that selective knockout of CD1d in neurons led to NDD development in mice. We also gathered biocomputational evidence from both human and mouse brains showing that neurons near the choroid plexus have the highest CD1d expression.

These preliminary findings inspired the NKT-in-NDD project with the following three aims:
(1) To determine whether CD1d-expressing neurons can sufficiently activate NKT cells
(2) To examine the impact of CD1d-dependent neuronal activation of NKT cells on inflammatory conditions and NDDs
(3) To verify whether CD1d is expressed by neurons localized near the choroid plexus

We have addressed and will continue to address various gaps in the fields of immune cell biology, neuroimmunology, and neuropathology. The neuroimmune insight gained from this project may expand current treatment options for neurodegenerative disease or other neuropathological ailments.

Work performed from the beginning of the project to the end of the period covered by the report and main results achieved so far

After the project start date, I got started right away with necessary trainings for project completion. I received training for flow cytometry, microscopy, and mouse handling from our core facilities. I was trained by fellow lab members in relevant laboratory techniques. These involved generating cultures from primary cortical neurons and splenocytes. I also learned how to generate panels for immune cell staining for relevant immune profiling. I also optimized the co-culturing experiments I performed, by enriching for NKT cells and finding the best incubation periods, cell numbers, and reagent concentrations. I also learned how to work with mice at many developmental stages, including new-born pups. I learned how to extract the spleen, brain, skull bone marrow, dura, and choroid plexus from these mice at various developmental stages, and how to maximize the immune cell detection for each immune niche.
I have also performed immunofluorescent staining of mouse brain sections to pinpoint the choroid plexus and have stained for immature neuronal markers and CD1d presence. I have learned how to perform whole-mount staining of the choroid plexus, in order to visualize the structural integrity and the immune milieu in situ.
I have also analyzed a number of public and in-house databases to localize CD1d expression in the brain and distinguish the role of CD1d in relevant cells, including neurons and choroid plexus epithelium. Collecting data from multiple databases from our lab and from other labs guarantees that my findings are specific and reproducible.
I have generated a number of microscopic images, .fcs files, and code, all of which will be uploaded to public repositories. I have optimized tissue collection and immune cell purification for future single-cell experiments which will be made publicly available.
Furthermore, I successfully co-cultured the primary cortical neurons and NKT cells, and stained for activation markers, showing that these CD1d-expressing neurons can indeed activate NKT cells. This is the first time that such a neuroimmune communication has been discovered and researched. All of my results will be shared via poster presentations and talks, both internally and externally. I also plan on publishing my results in an open-access journal and in bioRxiv.

Progress beyond the state of the art and expected potential impact (including the socio-economic impact and the wider societal implications of the project so far)

This project has identified a novel neuroimmune interaction never explored before. This could potentially inspire an interest in analyzing more neuroimmune interactions.
The project also identifies important neuroimmune niches, and allows for increased appreciation in their complexity and local milleus.
These two findings could inspire new insight in developing therapeutic intervention for various neuropathologies, including neurodegenerative disease or autoimmune diseases of the central nervous system.