Tobias Rogosch - Antibody repertoires in humanized NOD-scid-IL2Rγ(null) mice and human B cells reveals human-like diversification and tolerance checkpoints in the mouse.

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  Ippolito GC, Hoi KH, Reddy ST, Carroll SM, Ge X, Rogosch T,   Zemlin M, Shultz LD, Ellington AD, Vandenberg CL, Georgiou G.

  PLoS One.   2012;7(4):e35497.

  doi: 10.1371/journal.pone.0035497


  Immunodeficient mice reconstituted with human hematopoietic stem   cells enable the in vivo study of human hematopoiesis. In   particular, NOD-scid-IL2Rγ(null) engrafted mice have been shown   to have reasonable levels of T and B cell repopulation and can   mount T-cell dependent responses; however, antigen-specific   B-cell responses in this model are generally poor. We explored   whether developmental defects in the immunoglobulin gene   repertoire might be partly responsible for the low level of   antibody responses in this model. Roche 454 sequencing was used   to obtain over 685,000 reads from cDNA encoding immunoglobulin   heavy (IGH) and light (IGK and IGL) genes isolated from immature,   naïve, or total splenic B cells in engrafted NOD-scid-IL2Rγ(null)   mice, and compared with over 940,000 reads from peripheral B   cells of two healthy volunteers. We find that while naïve B-cell   repertoires in humanized mice are chiefly indistinguishable from   those in human blood B cells, and display highly correlated   patterns of immunoglobulin gene segment use, the   complementarity-determining region H3 (CDR-H3) repertoires are   nevertheless extremely diverse and are specific for each   individual. Despite this diversity, preferential D(H)-J(H)   pairings repeatedly occur within the CDR-H3 interval that are   strikingly similar across all repertoires examined, implying a   genetic constraint imposed on repertoire generation. Moreover,   CDR-H3 length, charged amino-acid content, and hydropathy are   indistinguishable between humans and humanized mice, with no   evidence of global autoimmune signatures. Importantly, however, a   statistically greater usage of the inherently autoreactive   IGHV4-34 and IGKV4-1 genes was observed in the newly formed   immature B cells relative to naïve B or total splenic B cells in   the humanized mice, a finding consistent with the deletion of   autoreactive B cells in humans. Overall, our results provide   evidence that key features of the primary repertoire are shaped   by genetic factors intrinsic to human B cells and are principally   unaltered by differences between mouse and human stromal   microenvironments.

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