Experiments reported herein demonstrate that activation of bovine B cells via surface immunoglobulin M (sIgM) cross-linking, analogous to T-cell independent (TI-2) antigenic stimulation, results in the expression of CD5. Interestingly, in the presence of CD40 ligand, sIgM-mediated induction of CD5 on B cells was inhibited. These findings indicate that activation of bovine B cells via B-cell receptor (BCR) cross-linking results in a CD5(+) B-cell phenotype and that CD40 signalling is inhibitory to this process. Analysis of cytokine mRNA indicates that bovine B cells constitutively express tumour necrosis factor-alpha (TNF-alpha) and interleukin (IL)-1 beta transcripts in vitro, while IL-10 mRNA expression is induced following sIgM cross-linking. IL-12 p40 transcripts were produced by B cells activated by CD40, but not by BCR, ligation. Analysis of cytokine receptor mRNA indicates that activation through CD40, in the presence or absence of IgM cross-linking, results in increased IL-4 receptor-alpha (IL-4R alpha), IL-13R alpha 1 and interferon-alpha receptor 1 (IFN-alpha R1) mRNA levels. Overall, these findings suggest that activation of bovine B cells through BCR cross-linking yields an activation phenotype that differs substantially from that of B cells activated through CD40.