A Germline-Encoded Structural Arginine Trap Underlies the Anti-DNA Reactivity of a Murine V Gene Segment

Autoimmunity may have its origins of early repertoire selection in developmental B cells. Such a primary repertoire is probably shaped by selecting B cells that can efficiently perform productive signaling, stimulated by self-antigens in the bone marrow, such as DNA. In support of that idea, we previously found a V segment from V(H)10 family that can form antibodies that bind to DNA independent of CDR3 usage. In this paper we designed four antibody fragments in a novel single-chain pre-BCR (scpre-BCR) format containing germinal V gene segments from families known to bind DNA (V(H)10) or not (V(H)4) connected to a murine surrogate light chain (SLC), lacking the highly charged unique region (UR), by a hydrophilic peptide linker. We also tested the influence of CDR2 on DNA reactivity by shuffling the CDR2 loop. The scpre-BCRs were expressed in bacteria. V(H)10 bearing scpre-BCR could bind DNA, while scpre-BCR carrying the V(H)4 segment did not. The CDR2 loop shuffling hampered V(H)10 reactivity while displaying a gain-of-function in the nonbinding V(H)4 germline. We modeled the binding sites demonstrating the conservation of a positivity charged pocket in the V(H)10 CDR2 as the possible cross-reactive structural element. We presented evidence of DNA reactivity hardwired in a V gene, suggesting a structural mechanism for innate autoreactivity. Therefore, while autoreactivity to DNA can lead to autoimmunity, efficiently signaling for B cell development is likely a trade-off mechanism leading to the selection of potentially autoreactive repertoires.

Automated summary

The study suggests that autoimmunity may have its origins in early repertoire selection in developmental B cells, shaping a primary repertoire by selecting B cells that can efficiently perform productive signaling. These B cells are stimulated by self-antigens such as DNA in the bone marrow, potentially leading to autoimmunity.