Rabinovich, Svetlana L. R. Powell, Rebecca W. B. Lindsay, Ross Yuan, Maoli Carpov, Alexei Wilson, Aaron Lopez, Mary W. Coleman, John Wagner, Denise Sharma, Palka Kemelman, Marina J. Wright, Kevin P. Seabrook, John Arendt, Heather Martinez, Jennifer DeStefano, Joanne J. Chiuchiolo, Maria L. Parks, Christopher rVSV-EnvG<sub>4</sub>-G<sub>6</sub> characterization. <p>(a) After a 24 hr infection, total infected Vero cell lysates were collected and proteins were separated by SDS polyacrylamide gel electrophoresis (SDS-PAGE). Blot was probed with anti-VSV-N and anti-VSV-G<sub>IN</sub> CT, which does not recognize G<sub>NJ</sub>. IN: rVSV-EnvG<sub>4</sub>-G<sub>6</sub><sup>IN</sup>; NJ: rVSV-EnvG<sub>4</sub>-G<sub>6</sub><sup>NJ</sup>. (b-d) Sucrose-gradient purified rVSV-EnvG<sub>4</sub>-G<sub>6</sub> particles were separated by SDS-PAGE. (b) Blots of 10<sup>6</sup> pfu of rVSV-EnvG<sub>4</sub>-G<sub>6</sub><sup>IN</sup> and 2.5×10<sup>6</sup> pfu rVSV-EnvG<sub>4</sub>-G<sub>6</sub><sup>NJ</sup> vectors were probed separately for EnvG using anti-gp120. (c) Blot of 10<sup>6</sup> pfu rVSV-EnvG<sub>4</sub>-G<sub>6</sub><sup>IN</sup> was probed with anti-VSV-G<sup>IN</sup> CT. (d) Blot of 10<sup>6</sup> pfu rVSV-EnvG<sub>4</sub>-G<sub>6</sub><sup>IN</sup> and rVSV-EnvG<sub>4</sub>-G<sub>6</sub><sup>NJ</sup> was probed with anti-VSV-G. (e) Replication kinetics of rVSV-EnvG<sub>4</sub>-G<sub>6</sub> and rVSV-G<sub>4</sub> viruses in Vero cells. Vero cells were infected in 6-well plates at an MOI of 0.1. At various intervals post infection, supernatant was collected from duplicate wells and virus was titrated. Plaques were counted manually after cell fixation.</p> G position;Significant quantities;immunogenic HIV;Conformationally Intact;humoral responses;novel vector;cytoplasmic tail regions;surface glycoprotein;G gene;regimen eliciting;Functional HIV;flow cytometry;EnvG functionality;IM VSV;surface EnvG;Env trimers;HIV vaccine platform;clade B Env immunogen;attenuating quantities;SIV challenge;stomatitis virus;growth attenuation;elisa;attenuated HIV;HIV immunogens;Elicits Potent Cellular;Envelope Trimers;Western Blot analysis;surface G;il;rhesus macaques;neutralizing Ab titers;IM electroporated plasmids encoding EnvG;safety concerns 2014-09-12
    https://plos.figshare.com/articles/figure/_rVSV_EnvG_4_G_6_characterization_/1169523
10.1371/journal.pone.0106597.g002