Genetic dissection of TLR9 reveals complex regulatory and cryptic proinflammatory roles in mouse lupus

In lupus, Toll-like receptor 7 (TLR7) and TLR9 mediate loss of tolerance to RNA and DNA, respectively. Yet, TLR7 promotes disease, while TLR9 protects from disease, implying differences in signaling. To dissect this ‘TLR paradox’, we generated two TLR9 point mutants (lacking either ligand (TLR9K51E) or MyD88 (TLR9P915H) binding) in lupus-prone MRL/lpr mice. Ameliorated disease of Tlr9K51E mice compared to Tlr9−/− controls revealed a TLR9 ‘scaffold’ protective function that is ligand and MyD88 independent. Unexpectedly, Tlr9P915H mice were more protected than both Tlr9K51E and Tlr9WT mice, suggesting that TLR9 also possesses ligand-dependent, but MyD88-independent, regulatory signaling and MyD88-mediated proinflammatory signaling. Triple-mixed bone marrow chimeras showed that TLR9–MyD88-independent regulatory roles were B cell intrinsic and restrained differentiation into pathogenic age-associated B cells and plasmablasts. These studies reveal MyD88-independent regulatory roles of TLR9, shedding light on the biology of endosomal TLRs. Endosomal TLR7 and TLR9 recognize RNA and DNA ligands, respectively, and both signal via MyD88 yet appear to play opposing roles in autoimmunity. Shlomchik and colleagues examine this TLR ‘paradox’, reporting that TLR9 has two protective functions, including an as yet unidentified additional MyD88-independent signaling pathway that confers protection against autoimmunity.