Abstract
High-parameter single-cell phenotyping has enabled in-depth classification and interrogation of immune cells, but to date has not allowed for glycan characterization. Here, we develop CyTOF-Lec as an approach to simultaneously characterize many protein and glycan features of human immune cells at the single-cell level. We implemented CyTOF-Lec to compare glycan features between different immune subsets from blood and multiple tissue compartments, and to characterize HIV-infected cell cultures. Using bioinformatics approaches to distinguish preferential infection of cellular subsets from viral-induced remodeling, we demonstrate that HIV upregulates the levels of cell-surface fucose and sialic acid in a cell-intrinsic manner, and that memory CD4+ T cells co-expressing high levels of fucose and sialic acid are highly susceptible to HIV infection. Sialic acid levels were found to distinguish memory CD4+ T cell subsets expressing different amounts of viral entry receptors, pro-survival factors, homing receptors, and activation markers, and to play a direct role in memory CD4+ T cells’ susceptibility to HIV infection. The ability of sialic acid to distinguish memory CD4+ T cells with different susceptibilities to HIV infection was experimentally validated through sorting experiments. Together, these results suggest that HIV remodels not only cellular proteins but also glycans, and that glycan expression can differentiate memory CD4+ T cells with vastly different susceptibility to HIV infection.
Original language | English (US) |
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Article number | e78870 |
Journal | eLife |
Volume | 11 |
DOIs | |
State | Published - Jul 2022 |
Funding
This work was supported by the National Institutes of Health R01AI127219, R01AI147777, and P01AI131374, UM1 AI164559, and UM1 AI164567 to NRR, and R01DK123733, R01AG062383, R01NS117458, and R21AI143385 to MAM. We also acknowledge NIH for the sorter (S10-RR028962), support from CFAR (P30AI027763), and the James B Pendleton Charitable Trust. We acknowledge the PFCC (RRID:SCR_018206) for assistance in CyTOF data acquisition, enabled by an instrument that was supported in part by the DRC Center Grant NIH P30 DK063720 and NIH S10 1S10OD018040-01. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. We thank Trimble Spitzer for the endometrial specimens; Nicole Lazarus and Eugene Butcher for the Act1 antibody; Claudia Bispo and Stanley Tamaki for CyTOF assistance at the Parnassus Flow Core; and Jane Srivastava and Nandhini Raman for flow cytometry assistance at the Gladstone Flow Core. We also thank Françoise Chanut for editorial assistance, and Robin Givens for administrative assistance.
ASJC Scopus subject areas
- General Neuroscience
- General Biochemistry, Genetics and Molecular Biology
- General Immunology and Microbiology