DOI: 10.5061/dryad.hmgqnk9rm
title: Supplementary information for: Macrophage- and CD4+ T cell-derived SIV differ in glycosylation, infectivity, and neutralization sensitivity

description: 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.
description: # Supplementary information for: Macrophage- and CD4+ T cell-derived SIV
 differ in glycosylation, infectivity, and neutralization sensitivity
 [https://doi.org/10.5061/dryad.hmgqnk9rm](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