Supplementary information for: Macrophage- and CD4+ T cell-derived SIV differ in glycosylation, infectivity, and neutralization sensitivity
Data files
Apr 24, 2024 version files 511.04 KB
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2A_Data.pzfx
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2A_Data.xlsx
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2BC_Data.pzfx
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2BC_Data.xlsx
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2D_Data.pzfx
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2D_Data.xlsx
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3AB_Data.pzfx
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3AB_Data.xlsx
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3CDE_Data.pzfx
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3CDE_Data.xlsx
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README.md
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S2_Data.pzfx
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S2_Data.xlsx
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Abstract
The human immunodeficiency virus (HIV) envelope protein (Env) mediates viral entry into host cells and is the primary target for the humoral immune response. Env is extensively glycosylated, and these glycans shield underlying epitopes from neutralizing antibodies. The glycosylation of Env is influenced by the type of host cell in which the virus is produced. Thus, HIV is distinctly glycosylated by CD4+ T cells, the major target cells, and macrophages. However, the specific differences in glycosylation between viruses produced in these cell types have not been explored at the molecular level. Moreover, it remains unclear whether the production of HIV in CD4+ T cells or macrophages affects the efficiency of viral spread and resistance to neutralization. To address these questions, we employed the simian immunodeficiency virus (SIV) model. Glycan analysis implied higher relative levels of oligomannose-type N-glycans in SIV from CD4+ T cells (T-SIV) compared to SIV from macrophages (M-SIV), and the complex-type N-glycans profiles seem to differ between the two viruses. Notably, M-SIV demonstrated greater infectivity than T-SIV, even when accounting for Env incorporation, suggesting that host cell-dependent factors influence infectivity. Further, M-SIV was more efficiently disseminated by HIV-binding cellular lectins. We also evaluated the influence of cell type-dependent differences on SIV's vulnerability to carbohydrate-binding agents (CBAs) and neutralizing antibodies. T-SIV demonstrated greater susceptibility to mannose-specific CBAs, possibly due to its elevated expression of oligomannose-type N-glycans. In contrast, M-SIV exhibited higher susceptibility to neutralizing sera in comparison to T-SIV. These findings underscore the importance of host cell-dependent attributes of SIV, such as glycosylation, in shaping both infectivity and the potential effectiveness of intervention strategies.
README: Supplementary information for: Macrophage- and CD4+ T cell-derived SIV differ in glycosylation, infectivity, and neutralization sensitivity
https://doi.org/10.5061/dryad.hmgqnk9rm
Description of the data and file structure
File names correspond to the figure in the manuscript for which the file supplies the underlying data. For instance, "2BC_Data" encompasses all data utilized in generating graphs for Fig. 2B and Fig. 2C. These data sets are stored as GraphPad Prism files and include details such as the number of biological replicates, raw data for each replicate, any data processing or statistical analysis performed, and the resulting graphs. Any exclusion of data points is accompanied by an explanation for this decision noted on the raw data sheets. Additionally, all data underlying the figures has been uploaded in Excel tables using file titles identical to the prism files.
Infectivity assays: Dataset 2A
Western blot with subsequent analysis using ImageJ: Dataset 2BC
Neutralization assays: Dataset 3A-E
Challenge experiment in rhesus macaques, RT-PCR: Dataset S2
The data processing strategy is included in the file for each dataset.
Abbreviations:
- Env: envelope protein
- GNA: galanthus nivalis agglutinin
- HIV: human immunodeficiency virus
- IU: infectious units
- MOI: multiplicity of infections
- M-SIV/T-SIV: pooled SIV produced in primary rhesus macaque macrophages/CD4+ T cells
- p27: capsid protein of SIV
- SIV: simian immunodeficiency virus
- UEA: ulex europaeus agglutinin, numbers in datafile
- S2: animal identifiers
- wpi: weeks post infection