Antibodies provide crucial protection against many pathogens. To induce plasma cells and the development of memory B cells, not just binding of antigen, but also other signals are known to be crucial. Toll-like receptors (TLR) are known to be expressed on human and murine B cells and can directly trigger their activation. Considering that little is known about porcine B cells, the present study isolated peripheral mononuclear cells and CD21 positive B cells followed by stimulation with TLR ligands. We also determined the expression levels of TLRs at the transcriptional level. We show that porcine B cell can induce upregulation of activation markers such as CCR7 and CD1, as well as proliferation after stimulation with TLR 2, 3, 4, 7, 8 and 9 ligands. These effects were not dependent on bystander cells as they were also observed with purified B cells. Interestingly, a particular strong reactivity was found with TLR2 and TLR8 ligands, although the corresponding TLR’s were not those with the highest expression. We also demonstrate that innate responses in B cells were observed with different B cell subsets defined on their IgM, CD21, CD1 and CD2 expression. This knowledge will be of value for defining strategies to manipulate B cells responses, in particular using vaccines.