info:eu-repo/semantics/article
Image-based Quantification of Direct Cell-to-cell Transmission of Bovine Viral Diarrhea Virus
Fecha
2019-08Registro en:
Merwaiss, Fernando; Alvarez, Diego Ezequiel; Image-based Quantification of Direct Cell-to-cell Transmission of Bovine Viral Diarrhea Virus; Bio-protocol; Bio-protocol; 9; 15; 8-2019; 1-10
2331-8325
CONICET Digital
CONICET
Autor
Merwaiss, Fernando
Alvarez, Diego Ezequiel
Resumen
Different viruses rely on direct cell-to-cell transmission to propagate infection within the infected host. Measuring this mode of transmission in cultured cells is often complicated by the contribution of cell free viruses to spread, and the difficulty to distinguish between primary infected cellsthat produce the virus and neighboring cells that are the target of spreading. Here, we present a protocol to quantify cell-to-cell transmission of the model pestivirus bovine viral diarrhea virus that is based on the co-culture of producer cells that are infected with a reporter virus expressing mCherry andtarget cells that stably express GFP. Spread of cell-free viruses is blocked by the presence of a neutralizing antibody in the cell culture medium, and cell-associated transmission is unequivocally quantified by numbering cells that are positive for both GFP and mCherry using automated analysis of fluorescence microscopy images.