Human remyelination promoting antibody inhibits apoptotic signaling and differentiation through Lyn kinase in primary rat oligodendrocytes

J. Watzlawik, E. Holicky, D. D. Edberg, D. L. Marks, A. E. Warrington, B. R. Wright, R. E. Pagano, M. Rodriguez

Research output: Contribution to journalArticle

41 Citations (Scopus)

Abstract

Purpose: Human remyelination promoting IgM mAbs target oligodendrocytes (OLs) and function in animal models of multiple sclerosis (MS). However, their mechanism of action is unknown. This study seeks to identify the cellular mechanism of action of a recombinant human IgM on OL survival. Methods: Binding of rHIgM22 to the surface of rat OLs was studied by co-localization with various markers. RHIgM22-mediated effects on apoptotic signaling in OLs, differentiation markers, and signaling molecules were detected by Western blotting and immunoprecipitation. Results: RHIgM22 colocalized with integrin β3 but not other integrin β-chains in OLs. Downstream of integrin β3 we identified Src family kinase (SFK) Lyn as a key player of rHIgM22-mediated actions in OLs. Lyn immunoprecipitated in a complex together with integrin αvβ3 and PDGFaR. Lyn expression was 9-fold up-regulated and Lyn activation was 3-fold higher in rHIgM22-treated OL cultures compared with controls. RHIgM22 inhibited apoptotic signaling by greater than 10-fold reduction of caspase-3 and capsase-9 cleavage and reduced by 4-fold expression of differentiation markers MBP and MOG in OLs. SFK inhibitors PP2 and SU6656 inhibited Lyn activity and restored caspase-cleavage in OLs. A human IgM that did not promote remyelination and medium were used as controls. Conclusions: rHIgM22 prevented apoptotic signaling and inhibited OL differentiation by Lyn implying that IgM-mediated remyelination is due to protection of OPC and OLs rather than promotion of OPC differentiation.

Original languageEnglish (US)
Pages (from-to)1782-1793
Number of pages12
JournalGLIA
Volume58
Issue number15
DOIs
StatePublished - Nov 15 2010

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Oligodendroglia
Phosphotransferases
Antibodies
Integrins
Immunoglobulin M
Differentiation Antigens
src-Family Kinases
Caspases
Immunoprecipitation
Caspase 3
Multiple Sclerosis
Animal Models
Western Blotting

Keywords

  • Demyelination
  • Fibronectin
  • Multiple sclerosis
  • Src family kinase

ASJC Scopus subject areas

  • Cellular and Molecular Neuroscience
  • Neurology

Cite this

Watzlawik, J., Holicky, E., Edberg, D. D., Marks, D. L., Warrington, A. E., Wright, B. R., ... Rodriguez, M. (2010). Human remyelination promoting antibody inhibits apoptotic signaling and differentiation through Lyn kinase in primary rat oligodendrocytes. GLIA, 58(15), 1782-1793. https://doi.org/10.1002/glia.21048

Human remyelination promoting antibody inhibits apoptotic signaling and differentiation through Lyn kinase in primary rat oligodendrocytes. / Watzlawik, J.; Holicky, E.; Edberg, D. D.; Marks, D. L.; Warrington, A. E.; Wright, B. R.; Pagano, R. E.; Rodriguez, M.

In: GLIA, Vol. 58, No. 15, 15.11.2010, p. 1782-1793.

Research output: Contribution to journalArticle

Watzlawik, J, Holicky, E, Edberg, DD, Marks, DL, Warrington, AE, Wright, BR, Pagano, RE & Rodriguez, M 2010, 'Human remyelination promoting antibody inhibits apoptotic signaling and differentiation through Lyn kinase in primary rat oligodendrocytes', GLIA, vol. 58, no. 15, pp. 1782-1793. https://doi.org/10.1002/glia.21048
Watzlawik, J. ; Holicky, E. ; Edberg, D. D. ; Marks, D. L. ; Warrington, A. E. ; Wright, B. R. ; Pagano, R. E. ; Rodriguez, M. / Human remyelination promoting antibody inhibits apoptotic signaling and differentiation through Lyn kinase in primary rat oligodendrocytes. In: GLIA. 2010 ; Vol. 58, No. 15. pp. 1782-1793.
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AU - Edberg, D. D.

AU - Marks, D. L.

AU - Warrington, A. E.

AU - Wright, B. R.

AU - Pagano, R. E.

AU - Rodriguez, M.

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N2 - Purpose: Human remyelination promoting IgM mAbs target oligodendrocytes (OLs) and function in animal models of multiple sclerosis (MS). However, their mechanism of action is unknown. This study seeks to identify the cellular mechanism of action of a recombinant human IgM on OL survival. Methods: Binding of rHIgM22 to the surface of rat OLs was studied by co-localization with various markers. RHIgM22-mediated effects on apoptotic signaling in OLs, differentiation markers, and signaling molecules were detected by Western blotting and immunoprecipitation. Results: RHIgM22 colocalized with integrin β3 but not other integrin β-chains in OLs. Downstream of integrin β3 we identified Src family kinase (SFK) Lyn as a key player of rHIgM22-mediated actions in OLs. Lyn immunoprecipitated in a complex together with integrin αvβ3 and PDGFaR. Lyn expression was 9-fold up-regulated and Lyn activation was 3-fold higher in rHIgM22-treated OL cultures compared with controls. RHIgM22 inhibited apoptotic signaling by greater than 10-fold reduction of caspase-3 and capsase-9 cleavage and reduced by 4-fold expression of differentiation markers MBP and MOG in OLs. SFK inhibitors PP2 and SU6656 inhibited Lyn activity and restored caspase-cleavage in OLs. A human IgM that did not promote remyelination and medium were used as controls. Conclusions: rHIgM22 prevented apoptotic signaling and inhibited OL differentiation by Lyn implying that IgM-mediated remyelination is due to protection of OPC and OLs rather than promotion of OPC differentiation.

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