Deliberate Attenuation of Chikungunya Virus by Adaptation to Heparan Sulfate-Dependent Infectivity: A Model for Rational Arboviral Vaccine Design (original) (raw)
Figure 6
Soluble heparin can block infectivity and alter plaque size of several of the positive charge CHIKV E2 mutants on MC3T3-E1 osteoblasts.
A) Soluble heparin was used to block infectivity of the virus by competing with the ability of the virus to attach to cells. Virus was incubated with 200 µg of either soluble heparin or BSA, infection was synchronized and overlay was added for a standard plaque assay. Percent heparin blocking was normalized to BSA control. B & C) Standard plaque assay on MC3T3-E1 was used to determine plaque size of the different CHIKV viruses. C) Plaque sizes were quantified by measuring the size of the plaques using a caliper. D) Virus was titered using a standard plaque assay on MC3T3-E1 cells in the presence or absence of 200 µg/mL of soluble heparin in the overlay. Plaque sizes were quantified by measuring the size of the plaques using a caliper. Viruses are color-coded based on control groups (orange) or MSD severity: little or no attenuation (purple), partial attenuation (blue), or highly attenuated (green) compared to CHIKV-LR. Error bars represent standard deviation. *p<0.001 compared to CHIKV-LR (B) or 103 mM NaCl (C).