Table 2.
Protein comparison of uninfected and infected monocyte EVs.
| Protein | Uninfected Monocyte Exosome (U937) | Infected Monocyte Exosome (U1) | |
|---|---|---|---|
| Controls | TGF-β binding protein 4 | +++ | - |
| Nucleolin | - | +++ | |
| HSP70 | + | + | |
| HSP90 | + | + | |
| Complement Protein C1 | + | + | |
| Complement Protein C3 | + | + | |
| Complement Protein C4A | + | + | |
| Complement Protein C5 | + | + | |
| Complement Protein C7 | + | - | |
| Complement Protein C8 | + | + | |
| Complement Protein C9 | + | + | |
| Complement Protein H | + | - | |
| RNA Binding Proteins | hnRNP A1 | - | ++ |
| hnRNP R | - | + | |
| hnRNP D0 | - | ++ | |
| hnRNP Q | - | + | |
| hnRNP K | - | + | |
| hnRNP U | - | + | |
| hnRNP DL | - | + | |
| hnRNP C1/2 | - | + | |
| hnRNP A1/B1 | - | + | |
| hnRNP A/B | - | + | |
| Histones | Histone H1 | - | + |
| Histone H2A | - | + | |
| Histone H2B | + | + | |
| Histone H3 | + | + | |
| Histone H4 | + | + | |
| Kinases | PKM | +++ | +++ |
| PGK1 | + | + | |
| NDK | + | + | |
| Int Kinase | + | + | |
| Adeno Kinase | + | - | |
| cAMP Kinase | + | - | |
| PI3K | + | - | |
| Xylulose Kinase | + | - | |
| BADDA | + | - | |
| Infected Cell Exosomes | PKR | - | + |
| HMGB1 | - | ++ | |
| Interleukin enhancer-BF3 | - | ++ | |
| FVEB1 | - | ++ | |
| T-Complex Protein | - | ++ | |
| Adenylosuccinate lyase | - | ++ | |
| Guanine nuc-binding protein | - | ++ | |
| Uninfected Cell Exosomes | Spondin-1 | ++ | - |
| ILGBP | ++ | - | |
| EMILIN-2 | ++ | - | |
| ATP-citrate synthase | ++ | - | |
| Proteoglycan 4 | + | - | |
| Exostosin-1 | + | - |
The 5-day old cell supernatants from U937 (uninfected) and U1 cells (HIV-1-infected) were harvested, treated with ExoMAX overnight, and centrifuged. The resulting pellet was run on an iodixanol density gradient, and the 10.8 fraction (exosome fraction) [36] was treated with NT80/82 overnight. The resulting pellet was treated was then prepared for mass spectrometry, and the resulting peptides were identified using Proteome Discoverer software. A table showing key protein differences between U937 and U1 EVs. HSPs are used as a control while differences between EVs are seen in RNA binding proteins, histones, kinases, and other proteins. The amount of protein in each type of EV was evaluated semi-quantitatively based on the number of peptide sequences identified. Proteins are denoted as being absent (“-“) or present (“+”). When the difference in the number of peptide sequences identified for a protein in each type of EV is less than 3×, both proteins are denoted as present (“+”). If a protein is increased between 3× and 6× in one EV type over the other, then the protein is denoted as enriched (“++”). If a protein is increased by at least 6× in one EV type over the other, then the protein is denoted as significantly enriched (“+++”).