Signalling protein complexes isolated from primary human skin-resident T cells can be analysed by Multiplex IP-FCM

Stephen E.P. Smith; Steven C. Neier; Tessa R. Davis; Mark R. Pittelkow; Diana Gil; Adam G. Schrum

doi:10.1111/exd.12362

Signalling protein complexes isolated from primary human skin-resident T cells can be analysed by Multiplex IP-FCM

Stephen E.P. Smith, Steven C. Neier, Tessa R. Davis, Mark R. Pittelkow, Diana Gil, Adam G. Schrum

Immunology

Research output: Contribution to journal › Article › peer-review

8 Scopus citations

Abstract

Studying signal transduction in skin-resident T cells (sr-T cells) can be limited by the small size of clinical biopsies. Here, we isolated sr-T cells from clinical samples and analysed signalling protein complexes by multiplex immunoprecipitation detected by flow cytometry (mIP-FCM). In samples from two independent donors, antigenic stimulation induced signalling proteins to join shared complexes that were observed in seven pairwise combinations among five proteins. This demonstrates that sr-T cells isolated from small clinical samples provide sufficient material for mIP-FCM-based analysis of signalling-induced protein complexes. We propose that this strategy may be useful for gaining improved mechanistic insight of sr-T cell signal transduction associated with dermatological disease.

Original language	English (US)
Pages (from-to)	272-273
Number of pages	2
Journal	Experimental Dermatology
Volume	23
Issue number	4
DOIs	https://doi.org/10.1111/exd.12362
State	Published - Apr 2014

Keywords

Multiplex technology
Protein-protein interaction
Signal transduction
Skin-resident T cell
T cell antigen receptor

ASJC Scopus subject areas

Biochemistry
Molecular Biology
Dermatology

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10.1111/exd.12362

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title = "Signalling protein complexes isolated from primary human skin-resident T cells can be analysed by Multiplex IP-FCM",

abstract = "Studying signal transduction in skin-resident T cells (sr-T cells) can be limited by the small size of clinical biopsies. Here, we isolated sr-T cells from clinical samples and analysed signalling protein complexes by multiplex immunoprecipitation detected by flow cytometry (mIP-FCM). In samples from two independent donors, antigenic stimulation induced signalling proteins to join shared complexes that were observed in seven pairwise combinations among five proteins. This demonstrates that sr-T cells isolated from small clinical samples provide sufficient material for mIP-FCM-based analysis of signalling-induced protein complexes. We propose that this strategy may be useful for gaining improved mechanistic insight of sr-T cell signal transduction associated with dermatological disease.",

keywords = "Multiplex technology, Protein-protein interaction, Signal transduction, Skin-resident T cell, T cell antigen receptor",

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AU - Smith, Stephen E.P.

AU - Neier, Steven C.

AU - Davis, Tessa R.

AU - Pittelkow, Mark R.

AU - Gil, Diana

AU - Schrum, Adam G.

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AB - Studying signal transduction in skin-resident T cells (sr-T cells) can be limited by the small size of clinical biopsies. Here, we isolated sr-T cells from clinical samples and analysed signalling protein complexes by multiplex immunoprecipitation detected by flow cytometry (mIP-FCM). In samples from two independent donors, antigenic stimulation induced signalling proteins to join shared complexes that were observed in seven pairwise combinations among five proteins. This demonstrates that sr-T cells isolated from small clinical samples provide sufficient material for mIP-FCM-based analysis of signalling-induced protein complexes. We propose that this strategy may be useful for gaining improved mechanistic insight of sr-T cell signal transduction associated with dermatological disease.

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Signalling protein complexes isolated from primary human skin-resident T cells can be analysed by Multiplex IP-FCM

Abstract

Keywords

ASJC Scopus subject areas

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