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  • Review Article
  • Published:

Adding new dimensions: towards an integrative understanding of HIV-1 spread

Nature Reviews Microbiology volume 12, pages 563–574 (2014)Cite this article

Subjects

Key Points

  • Standard two-dimensional (2D) cell culture systems do not take into account crucial parameters, such as tissue architecture and composition, the shear flow of body fluids and cellular communication and motility, which determine the efficiency of HIV-1 spread and thus disease progression. New developments in this field of research increasingly facilitate integrative ex vivo and in vivo analyses that take these factors into account.

  • Organotypic or synthetic 3D culture systems mimic the physiology of individual target organs to enable studies of select aspects of HIV-1 pathogenesis ex vivo. Parallel developments in the visualization of productively infected cells or individual virus particles facilitate the tracking of dynamic infection processes in real time.

  • Advanced live-cell imaging approaches to follow the transmission of fluorescent HIV-1 support the idea of cell–cell transmission as a highly effective mechanism of HIV-1 infection.

  • Small-animal models, such as BLT (bone marrow–liver–thymus) humanized mice, provide new opportunities to study and visualize HIV-1 spread and pathogenesis in vivo. Intravital microscopy was recently used for the first time to study retrovirus replication in vivo and provided support for an important role for cell–cell transmission in HIV-1 spread. This approach opens new avenues for gaining important insights into the mechanisms and dynamics of retroviral spread in the infected host.

  • Integrative approaches have helped to define the pathogenic principles of the HIV-1 accessory protein Nef and suggest that Nef is a modulator of HIV-1 target cell motility that may undermine host immune responses by altering immune cell communication.

Abstract

In vitro studies in primary or immortalized cells continue to be used to elucidate the essential principles that govern the interactions between HIV-1 and isolated target cells. However, until recently, substantial technical barriers prevented this information from being efficiently translated to the more complex scenario of HIV-1 spread in the host in vivo, which has limited our understanding of the impact of host physiological parameters on the spread of HIV-1. In this Review, we discuss the recent development of imaging approaches to visualize HIV-1 spread and the adaptation of these approaches to organotypic ex vivo models and animal models. We focus on new concepts, including the mechanisms and in vivo relevance of cell–cell transmission for HIV-1 spread and the function of the HIV-1 pathogenesis factor Nef, which have emerged from the application of these integrative approaches in complex cell systems.

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Figure 1: HIV-1 vaginal transmission and systemic dissemination.
Figure 2: Complex ex vivo cell systems for studies of HIV-1 spread.
Figure 3: Models for retroviral spread in vivo.
Figure 4: Model for the effects of HIV-1 Nef on T cell trafficking and induction of humoral immunity.

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Acknowledgements

T.R.M. was supported by US National Institutes of Health (NIH) grants R01 AI097052, R01 DA036298, P01 AI0178897 and P30 AI060354. T.T.M. was supported by a Massachusetts General Hospital Executive Committee on Research (MGH ECOR) Tosteson Postdoctoral Fellowship Award, NIH training grant T32 AI007387, Harvard University Center for AIDS Research (CFAR) grant NIH 5P30AI060354-09 and a pilot grant from the Center for Human Immunology (NIH U19 AI082630). O.T.F. and A.I. are supported by the Deutsche Forschungsgemeinschaft (grant numbers SFB 638 and SFB 1129, respectively). O.T.F. is a member of the CellNetworks Cluster of Excellence EXC81.

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Authors and Affiliations

  1. Department of Infectious Diseases, Integrative Virology, University Hospital Heidelberg, Im Neuenheimer Feld 324, Heidelberg, 69120, Germany

    Oliver T. Fackler & Andrea Imle

  2. German Center for Infection Research, Heidelberg University, Heidelberg, 69120, Germany

    Oliver T. Fackler

  3. The Center for Immunology and Inflammatory Diseases (CIID), Massachusetts General Hospital, Boston, 02114, Massachusetts, USA

    Thomas T. Murooka & Thorsten R. Mempel

  4. Harvard Medical School, Boston, 02115, Massachusetts, USA

    Thomas T. Murooka & Thorsten R. Mempel

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  1. Oliver T. Fackler
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  2. Thomas T. Murooka
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Correspondence to Oliver T. Fackler or Thorsten R. Mempel.

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PowerPoint slides

Glossary

Two-dimensional cell culture systems

(2D cell culture systems). Classical culture systems in which cells settle on a 2D plastic or glass surface.

Diffusive percolation

A term introduced to describe the movement of free virions through epithelia in analogy to the movement of water molecules through porous media.

Lymph node

An organized stromal environment where lymph is filtered and where most T cells and B cells initially encounter their cognate antigens for the induction of adaptive immune responses. Migratory dendritic cells and antigen enter lymph nodes through the afferent lymphatics, whereas most T cells enter from the bloodstream through high endothelial venules.

Productive infection

Infection that comprises all steps of the viral life cycle, including viral protein and particle production.

Latent infection

Infection in which the life cycle halts as an integrated provirus and the transcription of viral genes does not occur.

Organotypic cultures

Ex vivo cultures of tissue or tissue sections that have been obtained from HIV-negative donors in which physiological infection by HIV-1 can be simulated. When the tissue architecture of these organs is preserved, such cultures are referred to as organotypic.

3D cell culture models

Cell culture models that use an extracellular matrix to provide spacing and a complex 3D architecture.

Second harmonic generation

A nonlinear light-scattering process by which certain biological structures, including collagen fibres, double the frequency of light, thus turning — for example — 900 nm infrared light into 450 nm blue light. This phenomenon is useful to visualize structural tissue elements in multiphoton imaging studies without the need for labelling.

Virological synapses

Ordered assemblies of viral and host proteins at the interface between infected (that is, donor) and uninfected (that is, target) cells that facilitate the transfer of infectious virus. Virus transfer can occur via cell-free particles across the synaptic cleft but may also involve transport along cellular protrusions.

Cytonemes

Cellular extensions that protrude from one cell towards a neighbouring cell and enable transport of surface-bound virus particles.

Nanotubes

Cellular protrusions that form membrane bridges between donor and target cells, the lumens of which can allow for exchange of virus, along with cytoplasmic contents.

BLT humanized mouse model

(Bone marrow–liver–thymus humanized mouse model). Immunodeficient mice that have been engrafted with foetal human thymus, foetal liver and haematopoietic stem cells in order to recreate a functional human immune system in a small-animal model.

Uropods

The narrow trailing edges of polarized, migrating leukocytes, which differ from the cell front not only in shape and position but also in the organelles, cytoskeletal proteins and membrane molecules involved in motility, such as adhesion molecules and signalling receptors, that they contain.

Immunological synapses

The contact interfaces between antigen-presenting cells and T cells; they support adhesion, polarized signal transduction and T cell activation.

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Fackler, O., Murooka, T., Imle, A. et al. Adding new dimensions: towards an integrative understanding of HIV-1 spread. Nat Rev Microbiol 12, 563–574 (2014). https://doi.org/10.1038/nrmicro3309

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