1. Cell Biology

Actin-regulated Siglec-1 nanoclustering influences HIV-1 capture and virus-containing compartment formation in dendritic cells

  1. Enric Gutiérrez-Martínez
  2. Susana Benet Garrab
  3. Nicolas Mateos
  4. Itziar Erkiziac
  5. Jon Ander Nieto-Garai
  6. Maier Lorizate
  7. Kyra JE Borgman
  8. Carlo Manzo
  9. Felix Campelo
  10. Nuria Izquierdo-Useros  Is a corresponding author
  11. Javier Martinez-Picado  Is a corresponding author
  12. Maria F Garcia-Parajo  Is a corresponding author
  1. The Barcelona Institute of Science and Technology, Spain
  2. Hospital Universitari Germans Trias i Pujol, Spain
  3. IrsiCaixa AIDS Research Institute, Spain
  4. Universidad del País Vasco (UPV/EHU), Spain
  5. Universitat Central de Catalunya, Spain
https://doi.org/10.7554/eLife.78836
Share this article
Cite this article
  1. Enric Gutiérrez-Martínez
  2. Susana Benet Garrab
  3. Nicolas Mateos
  4. Itziar Erkiziac
  5. Jon Ander Nieto-Garai
  6. Maier Lorizate
  7. Kyra JE Borgman
  8. Carlo Manzo
  9. Felix Campelo
  10. Nuria Izquierdo-Useros
  11. Javier Martinez-Picado
  12. Maria F Garcia-Parajo
(2023)
Actin-regulated Siglec-1 nanoclustering influences HIV-1 capture and virus-containing compartment formation in dendritic cells
eLife 12:e78836.
https://doi.org/10.7554/eLife.78836
  2. Download BibTeX
  3. Download .RIS

Abstract

The immunoglobulin-like lectin receptor CD169 (Siglec-1) mediates the capture of HIV-1 by activated dendritic cells (DC) through binding to sialylated ligands. These interactions result in a more efficient virus capture as compared to resting DCs, although the underlying mechanisms are poorly understood. Using a combination of super-resolution microscopy, single particle tracking and biochemical perturbations we studied the nanoscale organization of Siglec-1 on activated DCs and its impact on viral capture and its trafficking to a single viral-containing compartment. We found that activation of DCs leads to Siglec-1 basal nanoclustering at specific plasma membrane regions where receptor diffusion is constrained by Rho-ROCK activation and formin-dependent actin polymerization. Using liposomes with varying ganglioside concentrations, we further demonstrate that Siglec-1 nanoclustering enhances the receptor avidity to limiting concentrations of gangliosides carrying sialic ligands. Binding to either HIV-1 particles or ganglioside-bearing liposomes lead to enhanced Siglec-1 nanoclustering and global actin rearrangements characterized by a drop in RhoA activity, facilitating the final accumulation of viral particles in a single sac-like compartment. Overall, our work provides new insights on the role of the actin machinery of activated DCs in regulating the formation of basal Siglec-1 nanoclustering, being decisive for the capture and actin-dependent trafficking of HIV-1 into the virus-containing compartment.

Data availability

All data generated or analysed during this study are included in the manuscript and supporting file; Source Data files have been provided for Figures 1-6 and for the corresponding figure supplements.

Article and author information

Author details

  1. Enric Gutiérrez-Martínez

    Institute of Photonic Sciences, The Barcelona Institute of Science and Technology, Barcelona, Spain
    Competing interests
    No competing interests declared.

  2. Susana Benet Garrab

    Infectious Diseases Department, Hospital Universitari Germans Trias i Pujol, Badalona, Spain
    Competing interests
    No competing interests declared.

  3. Nicolas Mateos

    Institute of Photonic Sciences, The Barcelona Institute of Science and Technology, Barcelona, Spain
    Competing interests
    No competing interests declared.

  4. Itziar Erkiziac

    IrsiCaixa AIDS Research Institute, Badalona, Spain
    Competing interests
    No competing interests declared.

  5. Jon Ander Nieto-Garai

    Universidad del País Vasco (UPV/EHU), Bilbao, Spain
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-8665-099X

  6. Maier Lorizate

    Universidad del País Vasco (UPV/EHU), Bilbao, Spain
    Competing interests
    No competing interests declared.

  7. Kyra JE Borgman

    Institute of Photonic Sciences, The Barcelona Institute of Science and Technology, Barcelona, Spain
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-7898-2911

  8. Carlo Manzo

    Facultat de Ciències i Tecnologia, Universitat Central de Catalunya, Vic, Spain
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-8625-0996

  9. Felix Campelo

    Institute of Photonic Sciences, The Barcelona Institute of Science and Technology, Barcelona, Spain
    Competing interests
    Felix Campelo, Reviewing editor, eLife.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-0786-9548

  10. Nuria Izquierdo-Useros

    IrsiCaixa AIDS Research Institute, Badalona, Spain
    For correspondence
    nizquierdo@irsicaixa.es
    Competing interests
    Nuria Izquierdo-Useros, declares patent applications related to the inhibition of viral interactions with Siglec-1. application reference EP23382072.9. The author declare no other competing interests.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-1039-1821

  11. Javier Martinez-Picado

    IrsiCaixa AIDS Research Institute, Badalona, Spain
    For correspondence
    jmpicado@irsicaixa.es
    Competing interests
    Javier Martinez-Picado, declares patent applications related to the inhibition of viral interactions with Siglec-1. application reference EP23382072.9. The author declare no other competing interests.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-4916-2129

  12. Maria F Garcia-Parajo

    Institute of Photonic Sciences, The Barcelona Institute of Science and Technology, Barcelona, Spain
    For correspondence
    maria.garcia-parajo@icfo.eu
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-6618-3944

Funding

European Research Council (788546)

  • Maria F Garcia-Parajo

Spanish National Plan for Scientific and Technical Research and Innovation (PID2020-117145RB-I00)

  • Nuria Izquierdo-Useros

Generalitat de Catalunya (2017SGR1000)

  • Maria F Garcia-Parajo

FUNDACIÓ Privada MIR-PUIG

  • Maria F Garcia-Parajo

Fundación Cellex

  • Maria F Garcia-Parajo

European Commission (754558)

  • Nicolas Mateos

Spanish National Plan for Scientific and Technical Research and Innovation (CEX2019-000910-S)

  • Maria F Garcia-Parajo

Spanish National Plan for Scientific and Technical Research and Innovation (PID2020-113068RB-I00 / 10.13039/501100011033)

  • Maria F Garcia-Parajo

Spanish National Plan for Scientific and Technical Research and Innovation (PID2019-109870RB-I00)

  • Javier Martinez-Picado

Spanish National Plan for Scientific and Technical Research and Innovation (PID2020-117405GB-100)

  • Maier Lorizate

Spanish National Plan for Scientific and Technical Research and Innovation (RYC-2017-22227)

  • Felix Campelo

Spanish National Plan for Scientific and Technical Research and Innovation (RYC-2015-17896)

  • Carlo Manzo

Spanish National Plan for Scientific and Technical Research and Innovation (PID2019-106232RB-I00/10.13039/501100011033)

  • Felix Campelo

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Copyright

© 2023, Gutiérrez-Martínez et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

Metrics

  • 1,149
    views
  • 216
    downloads
  • 7
    citations

Views, downloads and citations are aggregated across all versions of this paper published by eLife.

Download links

A two-part list of links to download the article, or parts of the article, in various formats.

Downloads (link to download the article as PDF)

Open citations (links to open the citations from this article in various online reference manager services)

Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)

  1. Enric Gutiérrez-Martínez
  2. Susana Benet Garrab
  3. Nicolas Mateos
  4. Itziar Erkiziac
  5. Jon Ander Nieto-Garai
  6. Maier Lorizate
  7. Kyra JE Borgman
  8. Carlo Manzo
  9. Felix Campelo
  10. Nuria Izquierdo-Useros
  11. Javier Martinez-Picado
  12. Maria F Garcia-Parajo
(2023)
Actin-regulated Siglec-1 nanoclustering influences HIV-1 capture and virus-containing compartment formation in dendritic cells
eLife 12:e78836.
https://doi.org/10.7554/eLife.78836

Share this article

https://doi.org/10.7554/eLife.78836

Categories and tags

Further reading

    1. Cell Biology
    2. Computational and Systems Biology

    Unbiased identification of cell identity in dense mixed neural cultures

    Sarah De Beuckeleer, Tim Van De Looverbosch ... Winnok H De Vos
    Research Article

    Induced pluripotent stem cell (iPSC) technology is revolutionizing cell biology. However, the variability between individual iPSC lines and the lack of efficient technology to comprehensively characterize iPSC-derived cell types hinder its adoption in routine preclinical screening settings. To facilitate the validation of iPSC-derived cell culture composition, we have implemented an imaging assay based on cell painting and convolutional neural networks to recognize cell types in dense and mixed cultures with high fidelity. We have benchmarked our approach using pure and mixed cultures of neuroblastoma and astrocytoma cell lines and attained a classification accuracy above 96%. Through iterative data erosion, we found that inputs containing the nuclear region of interest and its close environment, allow achieving equally high classification accuracy as inputs containing the whole cell for semi-confluent cultures and preserved prediction accuracy even in very dense cultures. We then applied this regionally restricted cell profiling approach to evaluate the differentiation status of iPSC-derived neural cultures, by determining the ratio of postmitotic neurons and neural progenitors. We found that the cell-based prediction significantly outperformed an approach in which the population-level time in culture was used as a classification criterion (96% vs 86%, respectively). In mixed iPSC-derived neuronal cultures, microglia could be unequivocally discriminated from neurons, regardless of their reactivity state, and a tiered strategy allowed for further distinguishing activated from non-activated cell states, albeit with lower accuracy. Thus, morphological single-cell profiling provides a means to quantify cell composition in complex mixed neural cultures and holds promise for use in the quality control of iPSC-derived cell culture models.

    1. Cell Biology

    Endocytic recycling is central to circadian collagen fibrillogenesis and disrupted in fibrosis

    Joan Chang, Adam Pickard ... Karl E Kadler
    Research Article

    Collagen-I fibrillogenesis is crucial to health and development, where dysregulation is a hallmark of fibroproliferative diseases. Here, we show that collagen-I fibril assembly required a functional endocytic system that recycles collagen-I to assemble new fibrils. Endogenous collagen production was not required for fibrillogenesis if exogenous collagen was available, but the circadian-regulated vacuolar protein sorting (VPS) 33b and collagen-binding integrin α11 subunit were crucial to fibrillogenesis. Cells lacking VPS33B secrete soluble collagen-I protomers but were deficient in fibril formation, thus secretion and assembly are separately controlled. Overexpression of VPS33B led to loss of fibril rhythmicity and overabundance of fibrils, which was mediated through integrin α11β1. Endocytic recycling of collagen-I was enhanced in human fibroblasts isolated from idiopathic pulmonary fibrosis, where VPS33B and integrin α11 subunit were overexpressed at the fibrogenic front; this correlation between VPS33B, integrin α11 subunit, and abnormal collagen deposition was also observed in samples from patients with chronic skin wounds. In conclusion, our study showed that circadian-regulated endocytic recycling is central to homeostatic assembly of collagen fibrils and is disrupted in diseases.

    1. Cell Biology

    Hypersensitive intercellular responses of endometrial stromal cells drive invasion in endometriosis

    Chun-Wei Chen, Jeffery B Chavez ... Bruce J Nicholson
    Research Article Updated

    Endometriosis is a debilitating disease affecting 190 million women worldwide and the greatest single contributor to infertility. The most broadly accepted etiology is that uterine endometrial cells retrogradely enter the peritoneum during menses, and implant and form invasive lesions in a process analogous to cancer metastasis. However, over 90% of women suffer retrograde menstruation, but only 10% develop endometriosis, and debate continues as to whether the underlying defect is endometrial or peritoneal. Processes implicated in invasion include: enhanced motility; adhesion to, and formation of gap junctions with, the target tissue. Endometrial stromal (ESCs) from 22 endometriosis patients at different disease stages show much greater invasiveness across mesothelial (or endothelial) monolayers than ESCs from 22 control subjects, which is further enhanced by the presence of EECs. This is due to the enhanced responsiveness of endometriosis ESCs to the mesothelium, which induces migration and gap junction coupling. ESC-PMC gap junction coupling is shown to be required for invasion, while coupling between PMCs enhances mesothelial barrier breakdown.