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. 2019 Mar 27;10:572. doi: 10.3389/fimmu.2019.00572

Figure 3.

Figure 3

Screening of Bcl-6/Bcl-xL transduced B cell cultures for PRRSV-specific antibodies. Antibody concentrations in pooled supernatants from 768 minipool transduced B cell cultures from a single pig (A). Following two week culture, supernatants were pooled and screened by IgG and IgA ELISA. Median fluorescence intensity (MFI) of PRRSV-infected MARC-145 cells stained using the 768 transduced B cell minipool culture supernatants (B). Each data point represents an individual minipool B cell supernatant. Flow cytometric staining of PRRSV-1 infected and uninfected cells by selected B cell minipool supernatants (C). Staining was assessed using a pan-porcine Ig secondary antibody (Pig Ig) and a porcine IgG specific secondary antibody (Pig IgG) and data presented by subtraction of the MFI of uninfected cells from the MFI of infected cells (ΔMFI). Each data point represents an individual minipool B cell supernatant and bars represent mean ± SD. Supernatants from five selected minipool B cell cultures were re-screened for PRRSV-specific antibodies by staining of mixture of PRRSV-1 infected cells and uninfected cells pre-labeled with Tag-It Violet (D). Assessment of PRRSV-1 neutralization by these selected transduced B cell minipool supernatants (E). Neutralization of PRRSV-1 Olot/91 was assessed by immunofluorescence staining.