The degree of polymerization and sulfation patterns in heparan sulfate are critical determinants of cytomegalovirus entry into host cells
Fig 2
Inhibition of HCMV growth by glycosaminoglycan derivatives.
Primary human foreskin fibroblasts (HFF) grown in 96 well plate were pretreated for one hour with 10 μM of 1) 6-O-desulfated Arixtra, 2) Unmodified Arixtra, 3) Heparin sodium salt from porcine intestinal mucosa (PIHSS), 4) Enoxaparin, or series of heparin oligosaccharide from enoxaparin: 5) dp2, 6) dp4, 7) dp6, 8) dp8, 9) dp10, 10) dp12, 11) dp14, 12) dp16, 13) dp18, 14) dp20 15) > dp20 or control (dH2O). Cells were infected with GFP tagged HCMV (Towne strain) virus at an MOI of 3.0 in the presence of the test glycosaminoglycans, which were maintained in the cell culture medium. At 5 days post-infection, cells were fixed and number of foci (GFP) was counted in triplicate well for each sample under an epifluorescent microscope. An average of >200 foci were present in mock-treated infected cells. Percent of viral GFP was calculated compared to virus only infected control (100% GFP expression). Results are representative of three independent replicates. Standard error of mean was plotted as error bars. Data was analyzed by ordinary one way ANOVA with multiple comparisons comparing the means of each test with control, and corrected using Dunnett’s post hoc test, showing significant differences among means (p <0.0001).