Nanomedicine is a promising discipline within nanotechnology, using nanoparticles for the improvement of drug delivery, diagnostics and therapy. Gold nanoparticles (GNPs) are commonly used in a variety of biomedical applications due to their highly beneficial properties. Nonetheless, the question remains regarding the possible adverse impact of GNPs on the immune cell function such is the immune response of B lymphocytes (also known as B cells). Our results showed that certain types of GNPs were able to directly interact with B cells in vitro and in vivo. Moreover, GNPs were able to modulate B cell immune function, resulting in attenuation of secretion of (pro-)inflammatory modulators as well as suppression of antigen-specific antibody production. Importantly, throughout the project, we demonstrated that GNPs are highly biocompatible with B cells. Our results provide important information that can help to evaluate the appropriateness of GNPs for biomedical applications and better predict their safety and success for future translation of GNP applications to clinical studies.