Ion-abrasion scanning electron microscopy reveals surface-connected tubular conduits in HIV-infected macrophages

Adam E. Bennett, Kedar Narayan, Dan Shi, Lisa M. Hartnell, Karine Gousset, Haifeng He, Bradley C. Lowekamp, Terry S. Yoo, Donald Bliss, Eric O. Freed, Sriram Subramaniam

Research output: Contribution to journalArticlepeer-review

Abstract

HIV-1-containing internal compartments are readily detected in images of thin sections from infected cells using conventional transmission electron microscopy, but the origin, connectivity, and 3D distribution of these compartments has remained controversial. Here, we report the 3D distribution of viruses in HIV-1-infected primary human macrophages using cryo-electron tomography and ion-abrasion scanning electron microscopy (IA-SEM), a recently developed approach for nanoscale 3D imaging of whole cells. Using IA-SEM, we show the presence of an extensive network of HIV-1-containing tubular compartments in infected macrophages, with diameters of ∼150-200 nm, and lengths of up to ∼5 μm that extend to the cell surface from vesicular compartments that contain assembling HIV-1 virions. These types of surface-connected tubular compartments are not observed in T cells infected with the 29/31 KE Gag-matrix mutant where the virus is targeted to multi-vesicular bodies and released into the extracellular medium. IA-SEM imaging also allows visualization of large sheet-like structures that extend outward from the surfaces of macrophages, which may bend and fold back to allow continual creation of viral compartments and virion-lined channels. This potential mechanism for efficient virus trafficking between the cell surface and interior may represent a subversion of pre-existing vesicular machinery for antigen capture, processing, sequestration, and presentation.

Original languageEnglish (US)
Article numbere1000591
JournalPLoS pathogens
Volume5
Issue number9
DOIs
StatePublished - Sep 2009
Externally publishedYes

ASJC Scopus subject areas

  • Parasitology
  • Microbiology
  • Immunology
  • Molecular Biology
  • Genetics
  • Virology

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