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. 2014 Jan 29;9(1):e87098. doi: 10.1371/journal.pone.0087098

Figure 2. rRABV-mICAM-1 infects primary murine splenocytes and B cells more efficiently than the parental virus, rRABV.

Figure 2

Naïve primary murine splenocytes were infected at an MOI of 5 with rRABV or rRABV-mICAM-1, or treated with media alone (mock-infected) for two days in-vitro. No additional mitogens were added to the culture to maintain the B cells and accessory splenocytes in a resting state similar to that in which they would exist in-vivo at the time of initial immunization. A) Representative gating strategy of total live splenocytes stained for intracellular RABV N as a marker for infection and for cell-surface expression of B220 as a maker for B cells. The percentage in the upper right quadrant is a representative example of the percentage of B220+ B cells infected with the rRABV-based vaccine. B) Percent RABV N+ cells in the total live lymphocyte gate. C) Percent RABV N+ cells in the B220+ cell population. D) Pretreatment of sucrose-purified rRABV-mICAM-1 with a neutralizing anti-ICAM-1 antibody significantly reduced infection of B cells in culture. To compare two groups of data, we used an unpaired, two-tailed Student's t test. (* p<0.05; **, p<0.01; ***, p<0.001)