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Neurosphere-derived multipotent precursors promote neuroprotection by an immunomodulatory mechanism

Abstract

In degenerative disorders of the central nervous system (CNS), transplantation of neural multipotent (stem) precursor cells (NPCs) is aimed at replacing damaged neural cells1,2. Here we show that in CNS inflammation, NPCs are able to promote neuroprotection by maintaining undifferentiated features and exerting unexpected immune-like functions. In a mouse model of chronic CNS inflammation, systemically injected adult syngeneic NPCs use constitutively activated integrins and functional chemokine receptors to selectively enter the inflamed CNS. These undifferentiated cells survive repeated episodes of CNS inflammation by accumulating within perivascular areas where reactive astrocytes, inflamed endothelial cells and encephalitogenic T cells produce neurogenic and gliogenic regulators. In perivascular CNS areas, surviving adult NPCs induce apoptosis of blood-borne CNS-infiltrating encephalitogenic T cells, thus protecting against chronic neural tissue loss as well as disease-related disability. These results indicate that undifferentiated adult NPCs have relevant therapeutic potential in chronic inflammatory CNS disorders because they display immune-like functions that promote long-lasting neuroprotection.

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Figure 1: aNPCs transplanted intravenously persist in inflamed perivascular areas of the CNS for up to three months after transplantation in R-EAE mice.
Figure 2: Stem cell regulators co-localize with intravenously injected aNPCs that persist in perivascular areas of the CNS in R-EAE mice.
Figure 3: aNPCs constitutively expressing VLA-4 and chemokine receptors accumulate around inflamed CNS microvessels in R-EAE mice.
Figure 4: aNPCs induce apoptosis of encephalitogenic T cells in vitro and in vivo.

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Acknowledgements

We wish to thank R. Galli, A. Gritti, M. Muzio, A. Vescovi and L. Ricci-Vitiani for discussions. We are grateful to F. Mavilio for critically discussing the manuscript. We acknowledge the technical help of S. Bach, S. Bucello, E. Butti, C. Covino, R. Molteni, A. Palini and C. Sciorati. S.P. is the recipient of a fellowship from the National Multiple Sclerosis Society (NMSS). L.Z. is the recipient of a fellowship from The Myelin Project (TMP). This work was supported in part by the Italian Minister of Health, the Italian Multiple Sclerosis Foundation (FISM), NMSS and TMP.

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Correspondence to Gianvito Martino.

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Reprints and permissions information is available at npg.nature.com/reprintsandpermissions. The authors declare no competing financial interests.

Supplementary information

Supplementary Figure S1

I.v.-injection of syngenic aNPCs reduces clinical relapses in R-EAE mice. (PDF 57 kb)

Supplementary Figure S2

I.v.-injected syngenic aNPCs persist within CNS perivascular areas of R-EAE mice. (PDF 13711 kb)

Supplementary Figure S3

Confocal images showing co-localization experiments performed in aNPC-transplanted R-EAE mice 106 dpi. (PDF 4000 kb)

Supplementary Figure S4

Perivascular CNS areas from R-EAE mice express stem cell regulators and contain activated microglia. (PDF 17263 kb)

Supplementary Figure S5

Stem cell regulators co-localize with i.v.-injected aNPCs persisting within perivascular CNS areas from R-EAE mice. (PDF 17596 kb)

Supplementary Figure S6

aNPCs constitutively express VLA-4 and adhere to VCAM-1 expressing CNS inflamed microvessels from C57Bl/6 mice with chronic progressive EAE. (PDF 555 kb)

Supplementary Figure S7

aNPCs express wide range of functional pro-inflammatory chemokine receptors. (PDF 81 kb)

Supplementary Figure S8

In vitro and in vivo analysis of CD3+ cells undergoing apoptosis. (PDF 11511 kb)

Supplementary Figure S9

Expression of regulators of stem cell proliferation and differentiation, immune molecules and trophic factors by aNPCs. (PDF 55 kb)

Supplementary Figures Legends

Full text legends to accompany the above Supplementary Figures. (DOC 39 kb)

Supplementary Video S1

Animated 3D reconstruction of a CNS perivascular area showing persistence of β-gal+ cells in close contact to endothelial cells producing BMP-4. (MOV 235 kb)

Supplementary Video S2

Animated 3D reconstruction of a CNS perivascular area showing persistence of β-gal+ cells in close contact with CNS-infiltrating CD45+ cells producing Noggin. (MOV 246 kb)

Supplementary Video S3

Animated 3D reconstruction of a β-gal+ cell expressing PSA-NCAM and incorporating BrdU 106 dpi. (MOV 96 kb)

Supplementary Video S4

Animated 3D reconstruction of a β-gal+ cell incorporating BrdU 106 dpi. (MOV 134 kb)

Supplementary Video Legends (DOC 37 kb)

Supplementary Methods

Additional descriptions of methods used in this study. (DOC 63 kb)

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Pluchino, S., Zanotti, L., Rossi, B. et al. Neurosphere-derived multipotent precursors promote neuroprotection by an immunomodulatory mechanism. Nature 436, 266–271 (2005). https://doi.org/10.1038/nature03889

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