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Human B-cell ontogeny in humanized NOD/SCID γcnull mice generates a diverse yet auto/poly- and HIV-1-reactive antibody repertoire

Genes & Immunity volume 13, pages 399–410 (2012)Cite this article

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Abstract

Characterization of the human antibody (Ab) repertoire in mouse models of the human immune system is essential to establish their relevance in translational studies. Single human B cells were sorted from bone marrow and periphery of humanized NOD/SCID γcnull (hNSG) mice at 8–10 months post engraftment with human cord blood-derived CD34+ stem cells. Human IG variable heavy (VH) and kappa (Vκ) genes were amplified, cognate VH–Vκ gene-pairs assembled as single-chain variable fragment-Fc Abs (scFvFcs) and functional studies were performed. Although overall distribution of VH genes approximated the normal human Ab repertoire, analysis of the VH-third complementarity-determining regions in the mature B-cell subset demonstrated an increase in length and positive charges, suggesting autoimmune characteristics. Additionally, >70% of Vκ sequences utilized Vκ4-1, a germline gene associated with autoimmunity. The mature B-cell subset-derived scFvFcs displayed the highest frequency of autoreactivity and polyspecificity, suggesting defects in checkpoint control mechanisms. Furthermore, these scFvFcs demonstrated binding to recombinant HIV envelope corroborating previous observations of poly/autoreactivity in anti-HIVgp140 Abs. These data lend support to the hypothesis that anti-HIV broadly neutralizing antibodies may be derived from auto/polyspecific Abs that escaped immune elimination and that the hNSG mouse could provide a new experimental platform for studying the origin of anti-HIV-neutralizing Ab responses.

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Acknowledgements

We thank the staff at the Animal Research Facility at Dana-Farber Cancer Institute (DFCI) for excellent maintenance and mice handling and Michael Waring at the Flow Cytometry core facility at Ragon Institute, Massachusetts General Hospital, Charlestown, MA, USA, for assistance with single B-cell sorting. Biostatistical computations performed by Amany Awad and the DFCI Biostatistical Core service (Sandra Lee and Yang Feng), technical guidance from Islay Campbell, ForteBio and assistance from Raymond Moniz of the Marasco Laboratory with the preparation of figures are appreciated. The HIV envelope reactive monoclonal antibodies (4E10 from Hermann Katinger and b12 from Dennis Burton) were obtained through the AIDS Research and Reference Reagent Program, Division of AIDS, NIAID, NIH (Germantown, MD, USA). This work was partially supported by NIH grants R21 AI091557 and UO1 AI0070343 to WAM Financial support provided by the National Foundation for Cancer Research to the Center for Therapeutic Antibody Engineering at DFCI (NFCR-CTAE) is acknowledged. Research support by the Center For AIDS Research at Harvard University (CFAR-HU) is also acknowledged.

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Author notes

  1. H Chang

    Present address: 3Current address: Department of Biochemistry, School of Basic Medicine, Hebei Medical University, Shijiazhuang, China.,

  2. S Geng

    Present address: 4Current address: Tian Guang Shi Biotechnology Co. LTD, Beijing, China.,

  3. H Chang and S Biswas: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Cancer Immunology and AIDS, Dana-Farber Cancer Institute, Boston, MA, USA

    H Chang, S Biswas, A S Tallarico, P T N Sarkis, S Geng, M M Panditrao, Q Zhu & W A Marasco

  2. Department of Medicine, Harvard Medical School, Boston, MA, USA

    H Chang, S Biswas, A S Tallarico, P T N Sarkis, S Geng, M M Panditrao, Q Zhu & W A Marasco

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Correspondence to W A Marasco.

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Chang, H., Biswas, S., Tallarico, A. et al. Human B-cell ontogeny in humanized NOD/SCID γcnull mice generates a diverse yet auto/poly- and HIV-1-reactive antibody repertoire. Genes Immun 13, 399–410 (2012). https://doi.org/10.1038/gene.2012.16

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