Activation of mouse B lymphocyte by proteins containing hexahistidine

Many recombinant proteins are produced as a fusion protein tagging hexahistidine, which has been used for studying their biological function both in vitro and in vivo. Unexpectedly, we observed activation of BALB/c mouse splenocytes when treated with hexahistidine-tagged recombinant proteins. This activation was hexahistidine-specific since the anti-pentahistidine antibody completely neutralized the effect, and the bovine serum albumin conjugated with the hexahistidine peptides also showed a similar activation effect. The B cells seemed to be the activated splenocytes, since the cell population stained with the anti-immunoglobulin antibody and anti-CD80 antibody was increased after the treatment. However, the activation signal by hexahistidine was insufficient to fully differentiate the B lymphocytes. This result suggests that caution must be taken in the use of hexahistidine-tagged recombinant proteins, due to their nonspecific activation of B lymphocyte.