Anti-DNA Heavy Chain Germline-Encoded or Somatically Mutated in ''Knock In'' Mice Targeted with a B Cell Deletion, Anergy, and Receptor Editing

To study the relative contributions of clonal deletion, clonal anergy, and receptor editing to tolerance induction in autoreactive B cells and their dependence on B cell receptor affinity, we have constructed “knock in” mice in which germline encoded or somatically mutated, rearranged anti-DNA heavy (H) chains were targeted to the H chain locus of the mouse. The targeted H chains were expressed on the vast majority of bone marrow (BM) and splenic B cells and were capable of Ig class switching and the acquisition of somatic mutations. A quantitative analysis of B cell populations in the BM as well as of J k utilization and DNA binding of hybridoma Abs suggested that immature B cell deletion and light (L) chain editing were the major mechanisms affecting tolerance. Unexpectedly, these mechanisms were less effective in targeted mice expressing the somatically mutated, anti-DNA H chain than in mice expressing the germline-encoded H chain, possibly due to the greater abundance of high affinity, anti-DNA immature B cells in the BM. Consequently, autoreactive B cells that showed features of clonal anergy could be recovered in the periphery of these mice. Our results suggest that clonal deletion and receptor editing are interrelated mechanisms that act in concert to eliminate autoreactive B cells from the immune system. Clonal anergy may serve as a back-up mechanism for central tolerance, or it may represent an intermediate step in clonal deletion. The Journal of Immunology, 1998, 161: 4634–4645.

[1]  J. Monroe,et al.  Characterization of anti-single-stranded DNA B cells in a non-autoimmune background. , 1997, Journal of immunology.

[2]  Y. Pewzner‐Jung,et al.  Expression of an anti-DNA-associated VH gene in immunized and autoimmune mice. , 1997, Journal of immunology.

[3]  B. Diamond,et al.  Light Chain Usage in Anti–double-stranded DNA B Cell Subsets: Role in Cell Fate Determination , 1997, The Journal of experimental medicine.

[4]  D. Nemazee,et al.  BCR ligation induces receptor editing in IgM+IgD- bone marrow B cells in vitro. , 1997, Immunity.

[5]  K. Rajewsky,et al.  B cell development under the condition of allelic inclusion. , 1997, Immunity.

[6]  M. Radic,et al.  Receptor editing, immune diversification, and self-tolerance. , 1996, Immunity.

[7]  M. Jackson,et al.  B cells are exquisitely sensitive to central tolerance and receptor editing induced by ultralow affinity, membrane-bound antigen , 1996, The Journal of experimental medicine.

[8]  M. Jourdan,et al.  Intrinsic B cell defects in NZB and NZW mice contribute to systemic lupus erythematosus in (NZB x NZW)F1 mice , 1996, The Journal of experimental medicine.

[9]  N. Klinman The "clonal selection hypothesis" and current concepts of B cell tolerance. , 1996, Immunity.

[10]  H. Zachau,et al.  The mouse immunoglobulin ϰ locus contains about 140 variable gene segments , 1996 .

[11]  K. Rajewsky Clonal selection and learning in the antibody system , 1996, Nature.

[12]  M. Wabl,et al.  A Quasi-Monoclonal Mouse , 1996, Science.

[13]  L. Wen,et al.  Murine lupus in the absence of αβ T cells , 1996 .

[14]  G. Nossal Clonal anergy of B cells: a flexible, reversible, and quantitative concept , 1996, The Journal of experimental medicine.

[15]  Y. Pewzner‐Jung,et al.  Structural elements controlling anti-DNA antibody affinity and their relationship to anti-phosphorylcholine activity. , 1996, Journal of immunology.

[16]  J. Monroe,et al.  Tolerance sensitivity of immature-stage B cells: can developmentally regulated B cell antigen receptor (BCR) signal transduction play a role? , 1996, Journal of immunology.

[17]  C. Goodnow Balancing immunity and tolerance: deleting and tuning lymphocyte repertoires. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[18]  Rolf M. Zinkernagel,et al.  Immunology Taught by Viruses , 1996, Science.

[19]  E. L. Prak,et al.  Immunoglobulin heavy chain gene replacement: a mechanism of receptor editing. , 1995, Immunity.

[20]  K. Rajewsky,et al.  Rearrangement of upstream DH and VH genes to a rearranged immunoglobulin variable region gene inserted into the DQ52‐JH region of the immunoglobulin heavy chain locus , 1995, European journal of immunology.

[21]  R. Hardy,et al.  The site and stage of anti-DNA B-cell deletion , 1995, Nature.

[22]  W. Anderson,et al.  Structure-function correlates of autoantibodies to nucleic acids. Lessons from immunochemical, genetic and structural studies. , 1994, Molecular immunology.

[23]  M. Shlomchik,et al.  The role of B cells in lpr/lpr-induced autoimmunity , 1994, The Journal of experimental medicine.

[24]  J. Cyster,et al.  Competition for follicular niches excludes self-reactive cells from the recirculating B-cell repertoire , 1994, Nature.

[25]  G. Cote,et al.  Direct sequencing of PCR products in agarose gel slices. , 1994, Nucleic acids research.

[26]  R. Hardy,et al.  Deletion and editing of B cells that express antibodies to DNA. , 1994, Journal of immunology.

[27]  A. Basten,et al.  Reduced life span of anergic self-reactive B cells in a double- transgenic model , 1994, The Journal of experimental medicine.

[28]  K. Rajewsky,et al.  Targeted insertion of a variable region gene into the immunoglobulin heavy chain locus. , 1993, Science.

[29]  M. Mcgrath,et al.  B cells are anergic in transgenic mice that express IgM anti‐DNA antibodies , 1993, European journal of immunology.

[30]  J Erikson,et al.  B lymphocytes may escape tolerance by revising their antigen receptors , 1993, The Journal of experimental medicine.

[31]  D. Nemazee,et al.  Receptor editing in self-reactive bone marrow B cells , 1993, The Journal of experimental medicine.

[32]  S. Camper,et al.  Receptor editing: an approach by autoreactive B cells to escape tolerance , 1993, The Journal of experimental medicine.

[33]  M. Cooke,et al.  Elimination of self-reactive B lymphocytes proceeds in two stages: Arrested development and cell death , 1993, Cell.

[34]  A. Rolink,et al.  Development of autoimmune disease in SCID mice populated with long-term "in vitro" proliferating (NZB x NZW)F1 pre-B cells , 1992, The Journal of experimental medicine.

[35]  B. Diamond,et al.  Induction of tolerance to an IgG autoantibody. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[36]  D. Tillman,et al.  Both IgM and IgG anti-DNA antibodies are the products of clonally selective B cell stimulation in (NZB x NZW)F1 mice , 1992, The Journal of experimental medicine.

[37]  M. Teitell,et al.  Failed self-tolerance and autoimmunity in IgG anti-DNA transgenic mice. , 1992, Journal of immunology.

[38]  B. Stollar Autoantibodies and autoantigens: a conserved system that may shape a primary immunoglobulin gene pool. , 1991, Molecular immunology.

[39]  B. Ledermann,et al.  Establishment of a germ-line competent C57BL/6 embryonic stem cell line. , 1991, Experimental cell research.

[40]  C. Goodnow,et al.  Elimination from peripheral lymphoid tissues of self-reactive B lymphocytes recognizing membrane-bound antigens , 1991, Nature.

[41]  C. Goodnow,et al.  Breakdown of self-tolerance in anergic B lymphocytes , 1991, Nature.

[42]  R. Fischel,et al.  Recurrent utilization of genetic elements in V regions of antinucleic acid antibodies from autoimmune mice. , 1991, Journal of immunology.

[43]  J D Kemp,et al.  Resolution and characterization of pro-B and pre-pro-B cell stages in normal mouse bone marrow , 1991, The Journal of experimental medicine.

[44]  S. Behar,et al.  Characterization of somatically mutated S107 VH11-encoded anti-DNA autoantibodies derived from autoimmune (NZB x NZW)F1 mice , 1991, The Journal of experimental medicine.

[45]  B. Diamond,et al.  A novel class of anti-DNA antibodies identified in BALB/c mice , 1991, The Journal of experimental medicine.

[46]  R. Hardy,et al.  Expression of anti-DNA immunoglobulin transgenes in non-autoimmune mice , 1991, Nature.

[47]  D. Pisetsky,et al.  Anti-DNA antibodies from autoimmune mice arise by clonal expansion and somatic mutation , 1990, The Journal of experimental medicine.

[48]  T. Ternynck,et al.  Two murine natural polyreactive autoantibodies are encoded by nonmutated germ-line genes. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

[49]  D. Nemazee,et al.  Clonal deletion of B lymphocytes in a transgenic mouse bearing anti-MHC class I antibody genes , 1989, Nature.

[50]  D. Webster,et al.  V region sequences of anti-DNA and anti-RNA autoantibodies from NZB/NZW F1 mice. , 1988, Journal of immunology.

[51]  S. Smith‐Gill,et al.  Altered immunoglobulin expression and functional silencing of self-reactive B lymphocytes in transgenic mice , 1988, Nature.

[52]  M. Gulley,et al.  Identification of a murine pan-T cell antigen which is also expressed during the terminal phases of B cell differentiation. , 1988, Journal of immunology.

[53]  M. Cygler,et al.  Antibodies to DNA , 1988, BioEssays : news and reviews in molecular, cellular and developmental biology.

[54]  G. M. Griffiths,et al.  Molecular events during maturation of the immune response to oxazolone , 1985, Nature.

[55]  L. Staudt,et al.  Inter- and intraclonal diversity in the antibody response to influenza hemagglutinin , 1985, The Journal of experimental medicine.

[56]  S. Rudikoff,et al.  Monoclonal antibodies to DNA and RNA from NZB/NZW F1 mice: antigenic specificities and NH2 terminal amino acid sequences. , 1984, Journal of immunology.

[57]  S. Rudikoff,et al.  Polymorphism in immunoglobulin heavy chains suggesting gene conversion. , 1982, Proceedings of the National Academy of Sciences of the United States of America.

[58]  U. Opitz,et al.  A new model for investigations of T-cell functions in mice: differential immunosuppressive effects of two monoclonal anti-Thy-1.2 antibodies. , 1982, Immunobiology.

[59]  I. Weissman,et al.  B220: a B cell-specific member of the T200 glycoprotein family , 1981, Nature.

[60]  L. Herzenberg,et al.  Localization of murine Ig-1b and Ig-1a (IgG 2a) allotypic determinants detected with monoclonal antibodies. , 1979, Molecular immunology.

[61]  Taql,et al.  Different joining region J elements of the murine K immunoglobulin light chain locus are used at markedly different frequencies , 1999 .

[62]  E. L. Prak,et al.  Editing disease-associated autoantibodies. , 1997, Immunity.

[63]  M. Radic,et al.  Genetic and structural evidence for antigen selection of anti-DNA antibodies. , 1994, Annual review of immunology.

[64]  M. Weigert Light Chain Editing in K-deficient Animals: A Potential Mechanism of B Cell Tolerance By Eline Luning Prak, Mary Trounstine,* Dennis Huszar,* , 1994 .

[65]  D. Eilat The measurement of anti-DNA activity in the sera of patients with systemic lupus erythematosus: theoretical and practical considerations. , 1989, Autoimmunity.

[66]  E. Weiler,et al.  Monoclonal anti‐allotype antibody towards BALB/c IgM Analysis of specificity and site of a V‐C crossover in recombinant strain BALB‐Igh‐Va/Igh‐Cb , 1987, European journal of immunology.

[67]  西 美幸 Preferential rearrangement of the immunoglobulin κ chain joining region J[κ1] and J[κ2] segments in mouse spleen DNA , 1986 .