Optimized methods for imaging membrane nanotubes between T cells and trafficking of HIV-1

Methods. 2011 Jan;53(1):27-33. doi: 10.1016/j.ymeth.2010.04.002. Epub 2010 Apr 9.

Abstract

A wide variety of cell types, including immune cells, have been observed to frequently interact via transient, long-distance membrane connections. However, considerable heterogeneity in their structure, mode of formation and functional properties has emerged, suggesting the existence of distinct subclasses. Open-ended tunneling nanotubes allow for the trafficking of cytoplasmic material, e.g. endocytic vesicles, or the transmission of calcium signals. Closed-ended membrane nanotubes do not seamlessly connect the cytoplasm between two interacting cells and a junction exists within the nanotube or where the nanotube meets a cell body. Recent live cell imaging suggested that membrane nanotubes between T cells could present a novel route for HIV-1 transmission. Here, we describe detailed protocols for observing membrane nanotubes and HIV-1 trafficking by live cell fluorescence microscopy.

Publication types

  • Research Support, Non-U.S. Gov't
  • Review

MeSH terms

  • Cell Culture Techniques
  • Cell Membrane Structures / ultrastructure*
  • Cell Tracking
  • Fibronectins / metabolism
  • Fixatives
  • Green Fluorescent Proteins / biosynthesis
  • Green Fluorescent Proteins / genetics
  • HIV-1 / physiology*
  • Humans
  • Immunological Synapses / ultrastructure
  • Jurkat Cells
  • Microscopy, Confocal
  • Pseudopodia / ultrastructure
  • Recombinant Fusion Proteins / biosynthesis
  • Recombinant Fusion Proteins / genetics
  • T-Lymphocytes / ultrastructure*
  • T-Lymphocytes / virology
  • gag Gene Products, Human Immunodeficiency Virus / biosynthesis
  • gag Gene Products, Human Immunodeficiency Virus / genetics

Substances

  • Fibronectins
  • Fixatives
  • Recombinant Fusion Proteins
  • gag Gene Products, Human Immunodeficiency Virus
  • Green Fluorescent Proteins