10 different compounds, including natural and synthetic polypeptides, proteins, polysaccharides, amino acids, and steroid hormones, were assayed for their capacity to form complexes with peritoneal exudate cell RNA. Only molecules carrying negatively charged groups were able to do so.
The formation of RNA-antigen complexes was unrelated to the immuno-potency of the "antigen," was not an enzyme-dependent reaction, did not require the synthesis of RNA following introduction of the antigen, did not seem to involve antigen-specific RNAs, was not specific for macrophages, since HeLa cells could be used as effectively, and occurred when purified RNA was mixed with antigen only in the presence of divalent cations. The complexes were very stable, once formed, but could be dissociated by exhaustive dialysis against buffers containing a chelating agent.
The macrophage RNA-antigen complex therefore appears to be a chelate between anionic groups on the two components. Based on the total absence of a relationship between immunogenicity and the capacity to form such complexes, as well as the nonspecific nature of complex formation at every level examined, it appears unlikely that RNA-antigen complexes play a physiologically significant role in immune induction.