Imaging the immunological synapse between dendritic cells and T cells

Markey, Kate A., Gartlan, Kate H., Kuns, Rachel D., MacDonald, Kelli P. A. and Hill, Geoffrey R. (2015) Imaging the immunological synapse between dendritic cells and T cells. Journal of Immunological Methods, 423 40-44. doi:10.1016/j.jim.2015.04.029

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Author Markey, Kate A.
Gartlan, Kate H.
Kuns, Rachel D.
MacDonald, Kelli P. A.
Hill, Geoffrey R.
Title Imaging the immunological synapse between dendritic cells and T cells
Journal name Journal of Immunological Methods   Check publisher's open access policy
ISSN 1872-7905
0022-1759
Publication date 2015-08-01
Year available 2015
Sub-type Article (original research)
DOI 10.1016/j.jim.2015.04.029
Open Access Status Not Open Access
Volume 423
Start page 40
End page 44
Total pages 5
Place of publication Amsterdam, Netherlands
Publisher Elsevier
Collection year 2016
Language eng
Formatted abstract
Immunological synapse formation between antigen-specific T cells and antigen presenting cells (APC) involves reorganization of the cellular cytoskeleton (polymerization of filamentous actin) and recruitment of adhesion molecules (e.g. LFA-1, ICAM-1). This engagement is critical for the generation of specific immune responses. Until recently, quantitative, high-throughput measurements of these interactions have not been possible. Instead, previous assessment was reliant on qualitative microscopy of live cells, where typically the APC is adhered to a surface and the suspended T cell is required to migrate to facilitate synapse formation. While this methodology can demonstrate the capacity for synapse formation, it cannot accommodate quantification of large numbers of interacting cell pairs, nor does it allow for statistically robust comparison between test conditions.

We have developed a method for assessing immunological synapse formation between purified ex vivo dendritic cells (DCs) and responder antigen-specific CD4+ T cells using imaging flow cytometry, allowing us to quantify LFA-1 and f-actin rearrangement at the interface between DC/T cell pairs. This novel application of imaging flow cytometry represents a major advance in dendritic cell function and immunological synapse research as it facilitates quantitative, high throughput analysis of the interaction between live, ex vivo DC and T cells.
Keyword Imaging flow cytometry
Amnis ImageStream
Dendritic cell
Transgenic T cell
Phalloidin
LFA-1
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: Official 2016 Collection
School of Medicine Publications
 
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