Regulation of Immunoglobulin Light Chain Isotype Expression

Gorman, James R.; Alt, Frederick W.

doi:10.1016/s0065-2776(08)60607-0

Cited by 69 publications

(61 citation statements)

References 343 publications

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“…In mice, the Igκ locus generates the majority of IgL-expressing B cells (32). The Vκ locus organization is distinct from that of the V H locus.…”

Section: Resultsmentioning

confidence: 99%

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Highly sensitive and unbiased approach for elucidating antibody repertoires

Lin

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

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Developing B lymphocytes undergo V(D)J recombination to assemble germ-line V, D, and J gene segments into exons that encode the antigen-binding variable region of Ig heavy (H) and light (L) chains. IgH and IgL chains associate to form the B-cell receptor (BCR), which, upon antigen binding, activates B cells to secrete BCR as an antibody. Each of the huge number of clonally independent B cells expresses a unique set of IgH and IgL variable regions. The ability of V(D)J recombination to generate vast primary B-cell repertoires results from a combinatorial assortment of large numbers of different V, D, and J segments, coupled with diversification of the junctions between them to generate the complementary determining region 3 (CDR3) for antigen contact. Approaches to evaluate in depth the content of primary antibody repertoires and, ultimately, to study how they are further molded by secondary mutation and affinity maturation processes are of great importance to the B-cell development, vaccine, and antibody fields. We now describe an unbiased, sensitive, and readily accessible assay, referred to as high-throughput genome-wide translocation sequencing-adapted repertoire sequencing (HTGTS-Repseq), to quantify antibody repertoires. HTGTS-Rep-seq quantitatively identifies the vast majority of IgH and IgL V(D)J exons, including their unique CDR3 sequences, from progenitor and mature mouse B lineage cells via the use of specific J primers. HTGTS-Rep-seq also accurately quantifies DJ H intermediates and V(D)J exons in either productive or nonproductive configurations. HTGTS-Rep-seq should be useful for studies of human samples, including clonal B-cell expansions, and also for following antibody affinity maturation processes. 1). In this process, the V, D, and J coding ends are generated as covalent hairpins that must be opened and that are often further processed, before being joined by classical nonhomologous end joining (2). Processing of V, D, J coding ends can involve generation of deletions or insertions of nucleotides at the junction regions (2), including the frequent de novo addition of nucleotides by the terminal deoxynucleotidyl transferase component of the V(D)J recombination process (3). Notably the V(D)J junctional region encodes a major antigen contact region of the antibody variable region, known as complementarity determining region 3 (CDR3), and thus these junctional diversification processes make a huge contribution to antibody diversity.The mouse IgH locus spans 2.7 megabases (Mb). There are 100s of V H s in the several megabase distal portion of the IgH, with the number varying substantially in certain mouse strains (4). The V H s lie ∼100 kb upstream from a 50-kb region containing 13 D H s, which is followed several kilobases downstream by a 2-kb region containing four J H s. The IgH constant region (C H ) exons lie downstream of the J H s. After assembly of a V H DJ H exon, transcription initiates upstream of the V H and terminates downstream of the C H exons, with V(D)J and C H portions being fused...

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“…In mice, the Igκ locus generates the majority of IgL-expressing B cells (32). The Vκ locus organization is distinct from that of the V H locus.…”

Section: Resultsmentioning

confidence: 99%

“…3B), which is slightly lower than the predicted 67:33 ratio (33). This small deviation might reflect the presence of nonproductive VJκ joins in Igλ-positive cells (32).…”

Section: Resultsmentioning

confidence: 99%

Highly sensitive and unbiased approach for elucidating antibody repertoires

Lin

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

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“…During lymphocyte development, V(D)J recombination is highly regulated in the contexts of lineage specificity, developmental-stage specificity, and allelic exclusion (4,5). In addition, developing lymphocytes use strategies that require productive V(D)J rearrangements for continued developmental progression (4,5).…”

Section: Uring B and T Cell Development The Ig And T Cell Receptormentioning

confidence: 99%

“…In addition, developing lymphocytes use strategies that require productive V(D)J rearrangements for continued developmental progression (4,5). For example, Ig heavy chain (IgH) variable-region genes are assembled in progenitor (pro) B cells via an ordered process in which D H -to-J H rearrangement occurs on both alleles before V H -to-DJ H rearrangement.…”

Section: Uring B and T Cell Development The Ig And T Cell Receptormentioning

confidence: 99%

Deletion of the RAG2 C terminus leads to impaired lymphoid development in mice

Akamatsu

Monroe

Dudley

et al. 2003

Proc. Natl. Acad. Sci. U.S.A.

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During lymphocyte development, V(D)J recombination is highly regulated in the contexts of lineage specificity, developmental-stage specificity, and allelic exclusion (4, 5). In addition, developing lymphocytes use strategies that require productive V(D)J rearrangements for continued developmental progression (4, 5). For example, Ig heavy chain (IgH) variable-region genes are assembled in progenitor (pro) B cells via an ordered process in which D H -to-J H rearrangement occurs on both alleles before V H -to-DJ H rearrangement. Productive V H -to-DJ H rearrangements produce cell-surface expression of IgH chains that signal expansion and differentiation of pro-B cells to the pre-B cell stage. This expression of IgH chains also signals the cessation of further V H -to-DJ H rearrangement via feedback regulation. Pro-B cells that first assemble nonproductive V H -to-DJ H rearrangements proceed to rearrange their second allele.Both RAG1 and RAG2 are required for V(D)J recombination, because RAG1-or RAG2-deficient mice exhibit a complete block in lymphocyte development at the progenitor stage (6, 7). In addition, RAG1 and RAG2 together are sufficient to initiate V(D)J recombination in vitro (8). Nearly all in vitro studies of RAG function have used the minimal regions of RAG1 (amino acids 384-1,008 of 1,040) and RAG2 (amino acids 1-383 of 527) required for activity, because full-length RAG1 and RAG2 are largely insoluble. However, the activities of these mutant ''core'' proteins differ from those of the full-length RAGs when assayed with extrachromosomal substrates in transfected cells. In this context, the mutant core RAG proteins support V(D)J recombination with reduced efficiency and with different levels and types of recombination products (9-15).Although not required for the biochemistry of V(D)J recombination, the noncore regions of RAG1 and RAG2 are conserved throughout evolution. For example, sequence conservation across the entire RAG2 protein from pufferfish to humans is Ϸ60%, with conservation of the noncore C-terminal region slightly higher than the core domain (16). Therefore, the noncore regions of RAG1 and RAG2 may serve important accessory and͞or regulatory functions in chromosomal V(D)J recombination. Consistent with this notion, studies of Abelson murine leukemia virus-transformed pre-B cells have suggested that the noncore regions of RAG1 are important for IgH locus D H -to-J H rearrangement (17), whereas the C terminus of RAG2 is more important for V H -to-DJ H rearrangement than for D H -to-J H rearrangement (18). The C terminus of RAG2 is predicted to fold into a plant homeodomain (PHD) (19,20), a motif found in a number of chromatin-associated proteins including some that have chromatin-modifying activities (21-24).To investigate potential in vivo function of the RAG2 noncore region in chromosomal V(D)J recombination and normal lymphocyte development, we have generated and characterized mice containing specific replacement of the full-length endogenous RAG2 gene with a gene encoding the mouse core...

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“…Two types of modeling theories have prevailed in an attempt to tackle this conundrum (3). The so-called stochastic models commonly considered a low probability p for an in-frame joint to occur due to inefficiency in either the onset or achievement of recombination (4)(5)(6).…”

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confidence: 99%