Exosomes are lipid nanovesicles released following fusion of the endosoma limiting membrane with the plasma membrane; however, their fate in lymphoid organs after their release remains controversial. We determined that sialoadhesin (CD169; Siglec-1) is required for the capture of B cell-derived exosomes via their surface-expressed α2,3-linked sialic acids. Exosome-capturing macrophages were present in the marginal zone of the spleen and in the subcapsular sinus of the lymph node. In vitro assays performed on spleen and lymph node sections confirmed that exosome binding to CD169 was not solely due to preferential fluid flow to these areas. Although the circulation half-life of exosomes in blood of wild-type and CD169(-/-) mice was similar, exosomes displayed altered distribution in CD169(-/-) mice, with exosomes freely accessing the outer marginal zone rim of SIGN-R1(+) macrophages and F4/80(+) red pulp macrophages. In the lymph node, exosomes were not retained in the subcapsular sinus of CD169(-/-) mice but penetrated deeper into the paracortex. Interestingly, CD169(-/-) mice demonstrated an enhanced response to antigen-pulsed exosomes. This is the first report of a role for CD169 in the capture of exosomes and its potential to mediate the immune response to exosomal antigen.