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Open Access Research

Analysis of detergent-free lipid rafts isolated from CD4+ T cell line: interaction with antigen presenting cells promotes coalescing of lipid rafts

Colleen Kennedy12, Matthew D Nelson13 and Anil K Bamezai1*

Author Affiliations

1 Department of Biology, Villanova University, 800 Lancaster Avenue, Villanova, PA 19085, USA

2 Current Address: Physician Assistant Program, Cornell Medical College, 1300 York Avenue, New York, NY 10065, USA

3 Current Address: Department of Neurology, University of Pennsylvania, 415 Curie Blvd., Philadelphia, PA 19104, USA

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Cell Communication and Signaling 2011, 9:31  doi:10.1186/1478-811X-9-31

Published: 8 December 2011

Abstract

Background

Lipid rafts present on the plasma membrane play an important role in spatiotemporal regulation of cell signaling. Physical and chemical characterization of lipid raft size and assessment of their composition before, and after cell stimulation will aid in developing a clear understanding of their regulatory role in cell signaling. We have used visual and biochemical methods and approaches for examining individual and lipid raft sub-populations isolated from a mouse CD4+ T cell line in the absence of detergents.

Results

Detergent-free rafts were analyzed before and after their interaction with antigen presenting cells. We provide evidence that the average diameter of lipid rafts isolated from un-stimulated T cells, in the absence of detergents, is less than 100 nm. Lipid rafts on CD4+ T cell membranes coalesce to form larger structures, after interacting with antigen presenting cells even in the absence of a foreign antigen.

Conclusions

Findings presented here indicate that lipid raft coalescence occurs during cellular interactions prior to sensing a foreign antigen.

Keywords:
raft coalescence; CD4+ T cells; antigen presenting cells; electron microscopy; raft-ELISA