Fig 3. M2-transduced B cells establish splenic plasma cell and germinal center B cell populations upon adoptive transfer in naïve mice.

Primary splenic B cells isolated by negative selection from naïve CD45.1+ donors were stimulated with LPS and subsequently transduced with M2- or M2.stop- retroviruses prior to adoptive transfer into naïve C57BL/6 mice (CD45.2+). Splenocytes from adoptive transfer recipients were harvested at D1 and D5 post transfer and cell surface markers were analyzed by flow cytometry. Data was compiled from two independent experiments with 3 mice/group. Each data point represents one animal and the horizontal bar represents the mean. Statistics were determined by two-tailed unpaired t test with Welch’s correction. (A) Representative flow plots depicting gating strategies to identify and subphenotype the transduced adoptively transferred B cell population (CD45.1+Thy1.1+) for plasma cell (B220^LoCD138^Hi) or germinal center B cell (B220+GL7^HiCD95^Hi) cell surface markers. Absolute numbers of splenic CD45.1+ B cells (B), CD45.1+Thy1.1+ B cells(C), CD45.1+B220+Thy1.1+ B cells (D), CD45.1+Thy1.1+ B cells displaying a plasma cell phenotype, and (E) CD45.1+B220+Thy1.1+ B cells displaying a germinal center phenotype (F).