Immunoglobulin heavy chain expression shapes the B cell receptor repertoire in human B cell development.

Developing B cells must pass a series of checkpoints that are regulated by membrane-bound Ig(mu) through the Igalpha-Igbeta signal transducers. To determine how Ig(mu) expression affects B cell development and Ab selection in humans we analyzed Ig gene rearrangements in pro-B cells from two patients who are unable to produce Ig(mu) proteins. We find that Ig(mu) expression does not affect V(H), D, or J(H) segment usage and is not required for human Igkappa and Iglambda recombination or expression. However, the heavy and light chains found in pro-B cells differed from those in peripheral B cells in that they showed unusually long CDR3s. In addition, the Igkappa repertoire in Ig(mu)-deficient pro-B cells was skewed to downstream Jkappas and upstream Vkappas, consistent with persistent secondary V(D)J rearrangements. Thus, Ig(mu) expression is not required for secondary V(D)J recombination in pro-B cells. However, B cell receptor expression shapes the Ab repertoire in humans and is essential for selection against Ab's with long CDR3s.

[1]  T. Waldmann,et al.  Developmental hierarchy of immunoglobulin gene rearrangements in human leukemic pre-B-cells. , 1981, Proceedings of the National Academy of Sciences of the United States of America.

[2]  T. Waldmann,et al.  Human immunoglobulin kappa light-chain genes are deleted or rearranged in lambda-producing B cells. , 1981, Nature.

[3]  T. Waldmann,et al.  Human immunoglobulin κ light-chain genes are deleted or rearranged in λ-producing B cells , 1981, Nature.

[4]  F. Alt,et al.  Ordered rearrangement of immunoglobulin heavy chain variable region segments. , 1984, The EMBO journal.

[5]  David Baltimore,et al.  Insertion of N regions into heavy-chain genes is correlated with expression of terminal deoxytransferase in B cells , 1984, Nature.

[6]  F. Alt,et al.  Activation of V kappa gene rearrangement in pre‐B cells follows the expression of membrane‐bound immunoglobulin heavy chains. , 1987, The EMBO journal.

[7]  R. Perlmutter,et al.  Early restriction of the human antibody repertoire. , 1987, Science.

[8]  D. Baltimore,et al.  Formation of disulphide-linked µ2ω2 tetramers in pre-B cells by the 18K ω-immunoglobulin light chain , 1987, Nature.

[9]  L. Hood,et al.  A T cell clone expresses two T cell receptor alpha genes but uses one alpha beta heterodimer for allorecognition and self MHC-restricted antigen recognition. , 1988, Cell.

[10]  D. Eisenberg,et al.  B29: a member of the immunoglobulin gene superfamily exclusively expressed on beta-lineage cells. , 1988, Proceedings of the National Academy of Sciences of the United States of America.

[11]  L. Hood,et al.  A T cell clone expresses two T cell receptor α genes but uses one αβ heterodimer for allorecognition and self MHC-restricted antigen recognition , 1988, Cell.

[12]  K. Rajewsky,et al.  A novel 34‐kd protein co‐isolated with the IgM molecule in surface IgM‐expressing cells. , 1988, The EMBO journal.

[13]  A. Carroll,et al.  Light-chain gene expression before heavy-chain gene rearrangement in pre-B cells transformed by Epstein-Barr virus. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

[14]  L. Hendershot,et al.  Mu heavy chains can associate with a pseudo-light chain complex (psi L) in human pre-B cell lines. , 1989, International immunology.

[15]  S. Levy,et al.  Activation of an excluded immunoglobulin allele in a human B lymphoma cell line. , 1989, Science.

[16]  D. Baltimore,et al.  Activation of immunoglobulin kappa gene rearrangement correlates with induction of germline kappa gene transcription , 1989, Cell.

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

[18]  S. Tonegawa,et al.  Junctional sequences of T cell receptor gamma delta genes: implications for gamma delta T cell lineages and for a novel intermediate of V-(D)-J joining. , 1989, Cell.

[19]  Joins,et al.  Isolation of scid pre‐B cells that rearrange kappa light chain genes: formation of normal signal and abnormal coding joins. , 1989, The EMBO journal.

[20]  Susumu Tonegawa,et al.  Junctional sequences of T cell receptor γδ genes: Implications for γδ T cell lineages and for a novel intermediate of V-(D)-J joining , 1989, Cell.

[21]  J. Y. Wang,et al.  Preferential utilization of conserved immunoglobulin heavy chain variable gene segments during human fetal life. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[22]  H. Karasuyama,et al.  The proteins encoded by the VpreB and lambda 5 pre-B cell-specific genes can associate with each other and with mu heavy chain , 1990, The Journal of experimental medicine.

[23]  P. Leder,et al.  Structure and expression of the human immunoglobulin lambda genes , 1990, The Journal of experimental medicine.

[24]  D. Tuveson,et al.  The products of pre-B cell-specific genes (lambda 5 and VpreB) and the immunoglobulin mu chain form a complex that is transported onto the cell surface , 1990, The Journal of experimental medicine.

[25]  M. Kopf,et al.  Molecular requirements for the mu‐induced light chain gene rearrangement in pre‐B cells. , 1991, The EMBO journal.

[26]  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.

[27]  K. Rajewsky,et al.  Most peripheral B cells in mice are ligand selected , 1991, The Journal of experimental medicine.

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

[29]  E. Kabat,et al.  Sequences of proteins of immunological interest , 1991 .

[30]  H. Boehmer,et al.  Kinetics and efficacy of positive selection in the thymus of normal and T cell receptor transgenic mice , 1991, Cell.

[31]  V. Pascual,et al.  Human immunoglobulin heavy-chain variable region genes: organization, polymorphism, and expression. , 1991, Advances in immunology.

[32]  Klaus Rajewsky,et al.  A B cell-deficient mouse by targeted disruption of the membrane exon of the immunoglobulin μ chain gene , 1991, Nature.

[33]  K. Rajewsky,et al.  A critical role of λ5 protein in B cell development , 1992, Cell.

[34]  Y. Uematsu,et al.  Exclusion and inclusion of α and β T cell receptor alleles , 1992, Cell.

[35]  E. Sasso,et al.  Emerging human B cell repertoire. Influence of developmental stage and interindividual variation. , 1992, Journal of immunology.

[36]  M. Reth,et al.  Crosslinking of the cell surface immunoglobulin (mu-surrogate light chains complex) on pre-B cells induces activation of V gene rearrangements at the immunoglobulin kappa locus. , 1992, International immunology.

[37]  T. Honjo,et al.  A transgenic model of autoimmune hemolytic anemia , 1992, The Journal of experimental medicine.

[38]  R. Schwartz,et al.  Immunoglobulin heavy chain gene expression in peripheral blood B lymphocytes. , 1992, The Journal of clinical investigation.

[39]  N. Berinstein,et al.  Up‐regulated recombination‐activating gene expression in slg− variants of a human mature B cell line undergoing secondary Igλ, rearrangements in cell culture , 1993, European journal of immunology.

[40]  C. Tonnelle,et al.  Mechanisms that generate human immunoglobulin diversity operate from the 8th week of gestation in fetal liver , 1993, European journal of immunology.

[41]  H. Zachau,et al.  Expressed human immunoglobulin ϰ genes and their hypermutation , 1993 .

[42]  A. Rolink,et al.  Rearrangement and expression of kappa light chain genes can occur without mu heavy chain expression during differentiation of pre-B cells. , 1993, International immunology.

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

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

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

[46]  H. Zachau,et al.  Expressed human immunoglobulin kappa genes and their hypermutation. , 1993, European journal of immunology.

[47]  K. Rajewsky,et al.  Immunoglobulin heavy and light chain genes rearrange independently at early stages of B cell development , 1993, Cell.

[48]  A. Rolink,et al.  Rearrangement and expression of χ light chain genes can occur without μ heavy chain expression during differentiation of pre-B cells , 1993 .

[49]  F. Alt,et al.  Immunoglobulin gene rearrangement in B cell deficient mice generated by targeted deletion of the JH locus. , 1993, International immunology.

[50]  K. Rajewsky,et al.  Human IgM+IgD+ B cells, the major B cell subset in the peripheral blood, express Vϰ genes with no or little somatic mutation throughout life , 1993, European journal of immunology.

[51]  D. Schatz,et al.  Down-regulation of RAG1 and RAG2 gene expression in preB cells after functional immunoglobulin heavy chain rearrangement. , 1995, Immunity.

[52]  J E Collins,et al.  Organization of the human immunoglobulin lambda light-chain locus on chromosome 22q11.2. , 1995, Human molecular genetics.

[53]  R. Brezinschek,et al.  Analysis of the heavy chain repertoire of human peripheral B cells using single-cell polymerase chain reaction. , 1995, Journal of immunology.

[54]  A. Glas,et al.  Polymorphism and Utilization of Human VH Genes a , 1995, Annals of the New York Academy of Sciences.

[55]  F. Ledeist,et al.  A human non-XLA immunodeficiency disease characterized by blockage of B cell development at an early proB cell stage. , 1996, The Journal of clinical investigation.

[56]  M. Lefranc,et al.  Sequence and evolution of the human germline V lambda repertoire. , 1996, Journal of molecular biology.

[57]  D. Campana,et al.  Mutations in the mu heavy-chain gene in patients with agammaglobulinemia. , 1996, The New England journal of medicine.

[58]  A. Rolink,et al.  Ordering of Human Bone Marrow B Lymphocyte Precursors by Single-Cell Polymerase Chain Reaction Analyses of the Rearrangement Status of the Immunoglobulin H and L Chain Gene Loci , 1996, The Journal of experimental medicine.

[59]  M. Milili,et al.  The VDJ repertoire expressed in human preB cells reflects the selection of bona fide heavy chains , 1996, European journal of immunology.

[60]  Michel C. Nussenzweig,et al.  Regulation of an Early Developmental Checkpoint in the B Cell Pathway by Igβ , 1996, Science.

[61]  E L Sonnhammer,et al.  Sequence of the human immunoglobulin diversity (D) segment locus: a systematic analysis provides no evidence for the use of DIR segments, inverted D segments, "minor" D segments or D-D recombination. , 1997, Journal of molecular biology.

[62]  A. Rolink,et al.  Changes in the V(H) gene repertoire of developing precursor B lymphocytes in mouse bone marrow mediated by the pre-B cell receptor. , 1997, Immunity.

[63]  F. Ledeist,et al.  A non-XLA primary deficiency causes the earliest known defect of B cell differentiation in humans: a comparison with an XLA case. , 1997, Immunology letters.

[64]  Sambasiva P Rao,et al.  The human heavy chain Ig V region gene repertoire is biased at all stages of B cell ontogeny, including early pre-B cells. , 1997, Journal of immunology.

[65]  R. Brezinschek,et al.  Analysis of the frequency and pattern of somatic mutations within nonproductively rearranged human variable heavy chain genes. , 1997, Journal of immunology.

[66]  R. Brezinschek,et al.  Molecular mechanisms and selective influences that shape the kappa gene repertoire of IgM+ B cells. , 1997, The Journal of clinical investigation.

[67]  M. Fischer,et al.  Molecular single-cell analysis reveals that CD5-positive peripheral blood B cells in healthy humans are characterized by rearranged Vkappa genes lacking somatic mutation. , 1997, The Journal of clinical investigation.

[68]  R. Brezinschek,et al.  Analysis of the human VH gene repertoire. Differential effects of selection and somatic hypermutation on human peripheral CD5(+)/IgM+ and CD5(-)/IgM+ B cells. , 1997, The Journal of clinical investigation.

[69]  I. Sanz,et al.  Human Ig heavy chain CDR3 regions in adult bone marrow pre-B cells display an adult phenotype of diversity: evidence for structural selection of DH amino acid sequences. , 1997, International immunology.

[70]  T. Manser,et al.  A Novel Mechanism for B Cell Repertoire Maturation Based on Response by B Cell Precursors to Pre–B Receptor Assembly , 1998, The Journal of experimental medicine.

[71]  A. Rolink,et al.  Precursor B cells showing H chain allelic inclusion display allelic exclusion at the level of pre-B cell receptor surface expression. , 1998, Immunity.

[72]  D. Campana,et al.  Mutations in the Human λ5/14.1 Gene Result in B Cell Deficiency and Agammaglobulinemia , 1998, The Journal of experimental medicine.

[73]  L. Rassenti,et al.  Chronic lymphocytic leukemia B cells express restricted sets of mutated and unmutated antigen receptors. , 1998, The Journal of clinical investigation.

[74]  M. Monestier,et al.  Atypical VH-D-JH rearrangements in newborn autoimmune MRL mice. , 1999, Journal of immunology.

[75]  J. Monroe,et al.  Negative selection of immature B cells by receptor editing or deletion is determined by site of antigen encounter. , 1999, Immunity.

[76]  C. Schaniel,et al.  Four of Five RAG-Expressing JCκ−/− Small Pre-BII Cells Have No L Chain Gene Rearrangements , 1999 .

[77]  Sambasiva P Rao,et al.  Biased VH gene usage in early lineage human B cells: evidence for preferential Ig gene rearrangement in the absence of selection. , 1999, Journal of immunology.

[78]  L. Bentolila,et al.  Extensive junctional diversity in Ig light chain genes from early B cell progenitors of mu MT mice. , 1999, Journal of immunology.

[79]  F. E. Bertrand,et al.  IgM heavy chain complementarity-determining region 3 diversity is constrained by genetic and somatic mechanisms until two months after birth. , 1999, Journal of immunology.

[80]  D. Campana,et al.  Mutations in Igα (CD79a) result in a complete block in B-cell development , 1999 .

[81]  C. Schaniel,et al.  Four of five RAG-expressing JCkappa-/- small pre-BII cells have no L chain gene rearrangements: detection by high-efficiency single cell PCR. , 1999, Immunity.

[82]  D. Campana,et al.  Mutations in Igalpha (CD79a) result in a complete block in B-cell development. , 1999, The Journal of clinical investigation.

[83]  K. Rajewsky,et al.  Rearrangement and Expression of Immunoglobulin Light Chain Genes Can Precede Heavy Chain Expression during Normal B Cell Development in Mice , 1999, The Journal of experimental medicine.

[84]  L. Staudt,et al.  Circulating human B cells that express surrogate light chains and edited receptors , 2000, Nature Immunology.

[85]  C. Janeway,et al.  A B-cell receptor-specific selection step governs immature to mature B cell differentiation. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[86]  P. Kirkham,et al.  Marriage, divorce, and promiscuity in human B cells , 2000, Nature Immunology.

[87]  E. Meffre,et al.  Autosomal primary immunodeficiencies affecting human bone marrow B‐cell differentiation , 2000, Immunological reviews.

[88]  Shiaoching Gong,et al.  B Cell Development Is Arrested at the Immature B Cell Stage in Mice Carrying a Mutation in the Cytoplasmic Domain of Immunoglobulin β , 2001, The Journal of experimental medicine.

[89]  I. Mårtensson,et al.  Loss of Precursor B Cell Expansion but Not Allelic Exclusion in VpreB1/VpreB2 Double-Deficient Mice , 2001, The Journal of experimental medicine.