scholarly journals Single-Molecule Super-Resolution Imaging of T-Cell Plasma Membrane CD4 Redistribution upon HIV-1 Binding

Viruses ◽  
2021 ◽  
Vol 13 (1) ◽  
pp. 142
Author(s):  
Yue Yuan ◽  
Caron A. Jacobs ◽  
Isabel Llorente Garcia ◽  
Pedro M. Pereira ◽  
Scott P. Lawrence ◽  
...  
Keyword(s):  
T Cells ◽  
Plasma Membrane ◽  
Single Molecule ◽  
Cd4 T Cells ◽  
Super Resolution ◽  
Fold Increase ◽  
Membrane Receptors ◽  
Virus Binding ◽  
Resolution Imaging ◽  
Hiv 1

The first step of cellular entry for the human immunodeficiency virus type-1 (HIV-1) occurs through the binding of its envelope protein (Env) with the plasma membrane receptor CD4 and co-receptor CCR5 or CXCR4 on susceptible cells, primarily CD4+ T cells and macrophages. Although there is considerable knowledge of the molecular interactions between Env and host cell receptors that lead to successful fusion, the precise way in which HIV-1 receptors redistribute to sites of virus binding at the nanoscale remains unknown. Here, we quantitatively examine changes in the nanoscale organisation of CD4 on the surface of CD4+ T cells following HIV-1 binding. Using single-molecule super-resolution imaging, we show that CD4 molecules are distributed mostly as either individual molecules or small clusters of up to 4 molecules. Following virus binding, we observe a local 3-to-10-fold increase in cluster diameter and molecule number for virus-associated CD4 clusters. Moreover, a similar but smaller magnitude reorganisation of CD4 was also observed with recombinant gp120. For one of the first times, our results quantify the nanoscale CD4 reorganisation triggered by HIV-1 on host CD4+ T cells. Our quantitative approach provides a robust methodology for characterising the nanoscale organisation of plasma membrane receptors in general with the potential to link spatial organisation to function.

scholarly journals Single-molecule super-resolution imaging of T-cell plasma membrane CD4 redistribution upon HIV-1 binding

2021 ◽  
Author(s):  
Yue Yuan ◽  
Caron Jacobs ◽  
Isabel Llorente Garcia ◽  
Pedro M. Pereira ◽  
Scott P. Lawrence ◽  
...  
Keyword(s):  
T Cells ◽  
Plasma Membrane ◽  
Single Molecule ◽  
Super Resolution ◽  
Fold Increase ◽  
Membrane Receptors ◽  
Host Cells ◽  
Virus Binding ◽  
Resolution Imaging ◽  
Hiv 1

The first step of cellular entry for the human immunodeficiency virus type-1 (HIV-1) occurs through the binding of its envelope protein (Env) with the plasma membrane receptor CD4 and co-receptor CCR5 or CXCR4 on susceptible cells, primarily CD4+ T cells and macrophages. Although there is considerable knowledge of the molecular interactions between Env and host cell receptors that lead to successful fusion, the precise way in which HIV-1 receptors redistribute to sites of virus binding at the nanoscale remains unknown. Here, we quantitatively examine changes in the nanoscale organisation of CD4 on the surface of CD4+ T cells following HIV-1 binding. Using single-molecule super-resolution imaging, we show that CD4 molecules are distributed mostly as either individual molecules or small clusters of up to 4 molecules. Following virus binding, we observe a local 3-to-10-fold increase in cluster diameter and molecule number for virus-associated CD4 clusters. Moreover, a similar but smaller magnitude reorganisation of CD4 was also observed with recombinant gp120. For the first time, our results quantify the nanoscale CD4 reorganisation triggered by HIV-1 on host cells. Our quantitative approach provides a robust methodology for characterising the nanoscale organisation of plasma membrane receptors in general with the potential to link spatial organisation to function.

scholarly journals Single molecule localisation microscopy reveals how HIV-1 Gag proteins sense membrane virus assembly sites in living host CD4 T cells

2018 ◽  
Vol 8 (1) ◽  
Author(s):  
Charlotte Floderer ◽  
Jean-Baptiste Masson ◽  
Elise Boilley ◽  
Sonia Georgeault ◽  
Peggy Merida ◽  
...  
Keyword(s):  
T Cells ◽  
Single Molecule ◽  
Cd4 T Cells ◽  
Virus Assembly ◽  
Gag Proteins ◽  
Localisation Microscopy ◽  
Hiv 1

scholarly journals The C-type lectin surface receptor DCIR acts as a new attachment factor for HIV-1 in dendritic cells and contributes to trans- and cis-infection pathways

Blood ◽  
2008 ◽  
Vol 112 (4) ◽  
pp. 1299-1307 ◽  
Author(s):  
Alexandra A. Lambert ◽  
Caroline Gilbert ◽  
Manon Richard ◽  
André D. Beaulieu ◽  
Michel J. Tremblay
Keyword(s):  
T Cells ◽  
Dendritic Cells ◽  
Cd4 T Cells ◽  
Productive Infection ◽  
Virus Binding ◽  
Virus Propagation ◽  
Cell Surface Molecule ◽  
Lectin Receptor ◽  
Trans Infection ◽  
Hiv 1

Abstract The dynamic interplay between dendritic cells (DCs) and human immunodeficiency virus type-1 (HIV-1) is thought to result in viral dissemination and evasion of antiviral immunity. Although initial observations suggested that the C-type lectin receptor (CLR) DC-SIGN was responsible for the trans-infection function of the virus, subsequent studies demonstrated that trans-infection of CD4+ T cells with HIV-1 can also occur through DC-SIGN–independent mechanisms. We demonstrate that a cell surface molecule designated DCIR (for DCimmunoreceptor), a member of a recently described family of DC-expressing CLRs, can participate in the capture of HIV-1 and promote infection in trans and in cis of autologous CD4+ T cells from human immature monocyte-derived DCs. The contribution of DCIR to these processes was revealed using DCIR-specific siRNAs and a polyclonal antibody specific for the carbohydrate recognition domain of DCIR. Data from transfection experiments indicated that DCIR acts as a ligand for HIV-1 and is involved in events leading to productive virus infection. Finally, we show that the neck domain of DCIR is important for the DCIR-mediated effect on virus binding and infection. These results point to a possible role for DCIR in HIV-1 pathogenesis by supporting the productive infection of DCs and promoting virus propagation.

scholarly journals HIV-1 Entry in SupT1-R5, CEM-ss, and Primary CD4+ T Cells Occurs at the Plasma Membrane and Does Not Require Endocytosis

2014 ◽  
Vol 88 (24) ◽  
pp. 13956-13970 ◽  
Author(s):  
N. Herold ◽  
M. Anders-Osswein ◽  
B. Glass ◽  
M. Eckhardt ◽  
B. Muller ◽  
...  
Keyword(s):  
T Cells ◽  
Plasma Membrane ◽  
Cd4 T Cells ◽  
Hiv 1

scholarly journals ThX – A next-generation probe for the early detection of amyloid aggregates

2019 ◽  
Author(s):  
Lisa-Maria Needham ◽  
Judith Weber ◽  
Juan A. Varela ◽  
James W. B. Fyfe ◽  
Dung T. Do ◽  
...  
Keyword(s):  
Fluorescent Probe ◽  
Single Molecule ◽  
Protein Misfolding ◽  
Super Resolution ◽  
Fold Increase ◽  
Next Generation ◽  
Aggregation Processes ◽  
Resolution Imaging ◽  
Localisation Microscopy ◽  
Protein Misfolding And Aggregation

AbstractNeurodegenerative diseases such as Alzheimer’s and Parkinson’s are associated with protein misfolding and aggregation. Recent studies suggest that the small, rare and heterogeneous oligomeric species, formed early on in the aggregation process, may be a source of cytotoxicity. Thioflavin T (ThT) is currently the gold-standard fluorescent probe for the study of amyloid proteins and aggregation processes. However, the poor photophysical and binding properties of ThT impairs the study of oligomers. To overcome this challenge, we have designed Thioflavin X, (ThX), a next-generation fluorescent probe which displays superior properties; including a 5-fold increase in brightness and 7-fold increase in binding affinity to amyloidogenic proteins. As an extrinsic dye, this can be used to study unique structural amyloid features both in bulk and on a single-aggregate level. Furthermore, ThX can be used as a super-resolution imaging probe in single-molecule localisation microscopy. Finally, we demonstrate that ThX can be used to detect a distinct oligomeric species, not observed via traditional ThT imaging.

Early proliferation of CCR5+ CD38+++ antigen-specific CD4+ Th1 effector cells during primary HIV-1 infection

Blood ◽  
2005 ◽  
Vol 106 (5) ◽  
pp. 1660-1667 ◽  
Author(s):  
John J. Zaunders ◽  
Mee Ling Munier ◽  
Daniel E. Kaufmann ◽  
Susanna Ip ◽  
Pat Grey ◽  
...  
Keyword(s):  
T Cells ◽  
Cd4 T Cells ◽  
Interleukin 2 ◽  
Specific Antigen ◽  
Fold Increase ◽  
Primary Response ◽  
T Helper 1 ◽  
Ki 67 ◽  
Effector Cells ◽  
Hiv 1

Abstract We investigated whether HIV-1 antigen-specific CD4+ T cells expressed the viral coreceptor CCR5 during primary HIV-1 infection (PHI). In the peripheral blood of subjects with very early PHI (< 22 days after onset of symptoms), there was a 10- to 20-fold increase in the proportion of highly activated (CD38+++) and proliferating (Ki-67+) CD4+ T cells that expressed CCR5+, and were mostly T-cell intracellular antigen-1 (TIA-1)+ perforin+ granzyme B+. Inthe same patient samples, CD4+ T cells producing interferon (IFN)–γ in response to HIV group-specific antigen (Gag) peptides were readily detected (median, 0.58%) by intracellular cytokine assay—these cells were again predominantly CD38+++, Ki-67+, and TIA-++, as well as Bcl-2low. On average, 20% of the Gag-specific CD4+ T cells also expressed interleukin-2 (IL-2) and were CD127 (IL-7R)+. Taken together, these results suggest that Gag-specific T-helper 1 (Th1) effector cells express CCR5 during the primary response and may include precursors of long-term self-renewing memory cells. However, in PHI subjects with later presentation, antigen-specific CD4+ T cells could not be readily detected (median, 0.08%), coinciding with a 5-fold lower level of the CCR5+CD38+++ CD4+ T cells. These results suggest that the antiviral response to HIV-1 infection includes highly activated CCR5+CD4+ cytotoxic effector cells, which are susceptible to both apoptosis and cytopathic infection with HIV-1, and rapidly decline.

scholarly journals Author Correction: Single molecule localisation microscopy reveals how HIV-1 Gag proteins sense membrane virus assembly sites in living host CD4 T cells

2018 ◽  
Vol 8 (1) ◽  
Author(s):  
Charlotte Floderer ◽  
Jean-Baptiste Masson ◽  
Elise Boilley ◽  
Sonia Georgeault ◽  
Peggy Merida ◽  
...  
Keyword(s):  
T Cells ◽  
Single Molecule ◽  
Cd4 T Cells ◽  
Virus Assembly ◽  
Gag Proteins ◽  
Localisation Microscopy ◽  
Hiv 1

scholarly journals Specific Lipid Recruitment by the Retroviral Gag Protein upon HIV-1 Assembly: From Model Membranes to Infected Cells

Proceedings ◽  
2020 ◽  
Vol 50 (1) ◽  
pp. 102
Author(s):  
Cyril Favard ◽  
Jakub Chojnacki ◽  
Naresh Yandrapalli ◽  
Johnson Mak ◽  
Christian Eggeling ◽  
...  
Keyword(s):  
T Cells ◽  
Plasma Membrane ◽  
In Silico ◽  
Cd4 T Cells ◽  
Virus Assembly ◽  
Gag Protein ◽  
Infected Cells ◽  
Specific Lipid ◽  
Hiv 1

The retroviral Gag protein targets the plasma membrane of infected cells for viral particle formation and release. The matrix domain (MA) of Gag is myristoylated for membrane anchoring but also contains a highly basic region that recognizes acidic phospholipids. Gag targets lipid molecules at the inner leaflet of the plasma membrane including phosphatidylinositol (4,5) bisphosphate (PI(4,5)P2) and cholesterol. Here, we addressed the question whether HIV-1 Gag was able to trap PI(4,5)P2 and/or other lipids during HIV-1 assembly in silico, in vitro on reconstituted membranes and in cellulo at the plasma membrane of the host CD4+ T cells. In silico, we could observe the first PI(4,5)P2 preferential recruitment by HIV-1 MA or Gag while protein docked on artificial membranes. In vitro, using biophysical techniques, we observed the specific trapping of PI(4,5)P2, and, to a lesser extent, cholesterol and the exclusion of sphingomyelin, during HIV-1 myr(-)Gag self-assembly on LUVs and SLBs. Finally, in infected living CD4+ T cells, we measured lipid dynamics within and away from HIV-1 assembly sites using super-resolution stimulated emission depletion (STED) microscopy coupled with scanning Fluorescence Correlation Spectroscopy (sSTED-FCS). The analysis of HIV-1 infected CD4+ T lymphocytes revealed that, upon virus assembly, HIV-1 is able to specifically trap PI(4,5)P2, and cholesterol but not phosphatidylethanolamine (PE) or sphingomyelin (SM) at the cellular membrane. Furthermore, analyzing CD4+ T cells expressing only HIV-1 Gag protein showed that Gag is the main driving force restricting the mobility of PI(4,5)P2 and cholesterol at the cell plasma membrane. Our data provide the first direct evidence showing that HIV-1 Gag creates its own specific lipid environment for virus assembly by selectively recruiting lipids to generate PI(4,5)P2/cholesterol-enriched nanodomains favoring virus assembly, and that HIV-1 does not assemble on pre-existing lipid domains.

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