Fig. 5. The pr-specific antibody 2A7 inhibits BinJV in an isotype-dependent manner.

(A) Purified 2A7 in Fab, IgG, and IgM form analyzed by reduced 4 to 12% SDS-PAGE. Heavy (HC) and light chains (LC) indicated by arrows. (B) Western blot probing with recombinant 2A7-Fab (before HRV3C digestion), IgG, and IgM. Purified BinJV from infected C6/36 cells was resolved by nonreducing 4 to 12% SDS-PAGE. (C) Neutralization of BinJV by 2A7-Fab, IgG, and IgM antibodies or anti-influenza CO5 antibody as isotype controls. Neutralization curves were determined by TCID₅₀ on C6/36 cells. Means ± range. OD, optical density. (D) Neutralization by 2A7-IgM or control CO5-IgM of BinJV produced in C6/36 (A. albopictus), RML12 (A. albopictus), and Aag2 (A. aegypti) mosquito cells with 2A7-IgM or control CO5-IgM. Neutralization was determined by TCID₅₀ on C6/36 cells. Means ± range. (E) Viral titers in A. albopictus mosquitoes injected intrathoracically with BinJV:2A7-IgM or BinJV:CO5-IgM. BinJV supernatant was incubated with either antibody (2A7-IgM or CO5-IgM) at 28°C for 1 hour before injection. Each point represents a single infected mosquito; means ± SD. Statistical analysis performed by unpaired t test with Welch’s correction. ***P ≤ 0.001.