scholarly journals Chromosomal position of a VH gene segment determines its activation and inactivation as a substrate for V(D)J recombination

2007 ◽  
Vol 204 (13) ◽  
pp. 3247-3256 ◽  
Author(s):  
Jamie Geier Bates ◽  
Dragana Cado ◽  
Hector Nolla ◽  
Mark S. Schlissel
Keyword(s):  
B Cells ◽  
Gene Cluster ◽  
Gene Targeting ◽  
Dna Sequences ◽  
Gene Rearrangement ◽  
Gene Segment ◽  
Allelic Exclusion ◽  
Chromosomal Position ◽  
Lineage Specificity ◽  
Vh Gene

Complete IgHC gene rearrangement occurs only in B cells in a stage-specific and ordered manner. We used gene targeting to reposition a distal VH gene segment to a region just 5′ of the DH gene cluster and found its activation to be highly dependent on the chromosomal domain within which it resides. The targeted VH gene segment rearranged at a higher frequency than its endogenous counterpart, its rearrangement was no longer ordered, and its ability to be silenced by allelic exclusion was lost. Additionally, the targeted VH gene segment lost lineage specificity, as VDJH rearrangement was observed in thymocytes. These data suggest that locus contraction, mimicked by proximal targeting, can override any regulation imposed by DNA sequences immediately surrounding VH gene segments.

scholarly journals Virus-transformed pre-B cells show ordered activation but not inactivation of immunoglobulin gene rearrangement and transcription.

1991 ◽  
Vol 173 (3) ◽  
pp. 711-720 ◽  
Author(s):  
M S Schlissel ◽  
L M Corcoran ◽  
D Baltimore
Keyword(s):  
B Cells ◽  
Cell Lines ◽  
Gene Rearrangement ◽  
Gene Segment ◽  
Allelic Exclusion ◽  
Immunoglobulin Gene ◽  
Chain Reaction ◽  
Highly Sensitive ◽  
Sensitive Polymerase Chain Reaction ◽  
Polymerase Chain

Virus-transformed pre-B cells undergo ordered immunoglobulin (Ig) gene rearrangements during culture. We devised a series of highly sensitive polymerase chain reaction assays for Ig gene rearrangement and unrearranged Ig gene segment transcription to study both the possible relationship between these processes in cultured pre-B cells and the role played by heavy (H) chain (mu) protein in regulating gene rearrangement. Our analysis of pre-B cell cultures representing various stages of maturity revealed that transcription of each germline Ig locus precedes or is coincident with its rearrangement. Cell lines containing one functional rearranged H chain allele, however, continue to transcribe and to rearrange the allelic, unrearranged H chain locus. These cell lines appear to initiate but not terminate rearrangement events and therefore provide information about the requirements for activating rearrangement but not about allelic exclusion mechanisms.

scholarly journals Immunoglobulin gene rearrangements in normal mouse B cells

1982 ◽  
Vol 2 (7) ◽  
pp. 829-836
Author(s):  
P Early ◽  
C Nottenburg ◽  
I Weissman ◽  
L Hood
Keyword(s):  
B Cells ◽  
Heavy Chain ◽  
Germ Line ◽  
Gene Segment ◽  
Immunoglobulin M ◽  
Chain Gene ◽  
Allelic Exclusion ◽  
Immunoglobulin Gene ◽  
Spleen Cells ◽  
Surface Immunoglobulin

We have analyzed the structure of rearranged mu heavy-chain genes obtained from the genomic DNA of normal BALB/c mouse spleen cells expressing surface immunoglobulin M. Examples were found of two types of nonproductive rearrangements, which may be responsible for allelic exclusion in normal B cells. In one of these rearrangements, a germ line D gene segment has joined to the JH4 gene segment but no V/D joining has occurred. We present evidence that D gene segments lie as a cluster between V and J gene segments in the germ line. A comparison of conserved sequences in V and D gene segments suggests that the D gene segments, which are found only in the heavy-chain gene family, may have evolved from V gene segments similar to the Vk family.

scholarly journals High-frequency representation of a single VH gene in the expressed human B cell repertoire.

1993 ◽  
Vol 177 (2) ◽  
pp. 409-418 ◽  
Author(s):  
A K Stewart ◽  
C Huang ◽  
B D Stollar ◽  
R S Schwartz
Keyword(s):  
B Cells ◽  
B Cell ◽  
High Frequency ◽  
Gene Segment ◽  
Normal Subjects ◽  
Cell Repertoire ◽  
B Cell Repertoire ◽  
V Genes ◽  
Vh Gene ◽  
Vh Genes

Idiotype (Id) 16/6 marks a variable (V) region structure that occurs frequently in the human immunoglobulin repertoire. The basis of the Id has been traced to a germline heavy chain gene segment, VH18/2 (VH26). To pursue the molecular basis for the frequency of Id 16/6, we have analyzed polymerase chain reaction-generated C mu, C gamma, and VH3 family V gene libraries derived from the circulating and tonsillar B cells of four normal individuals and from the B cells of two patients with active systemic lupus erythematosus (SLE). The frequency of VH18/2 in these libraries was compared with three control VH genes, VH56P1, VH21/28, and VHA57. Plaque lifts from C mu and C gamma VH cDNA libraries were screened with gene-specific oligonucleotide probes. The frequency of VH18/2 ranged from 4 to 10% of JH+ plaques (two of five times that of control VH genes). In four VH3 family-specific libraries derived from rearranged DNA, VH18/2 represented 19-33% of VH3+ plaques. Hybridizing VH18/2 plaques were 98-100% homologous to the germline VH gene; mutations when present were often in framework 3. Extensive variation was seen in the complementarity determining region 3 sequences of these rearranged V genes. The high frequency of VH18/2 expression in the B cell repertoire was confirmed by sequencing randomly picked JH+ plaques. In two patients with active SLE the frequency of use of VH18/2 was not greater than that observed in normal subjects. These results show that VH18/2 is overrepresented in the B cell repertoire of normal subjects and suggest that the immune repertoire may be dominated by relatively few V genes.

scholarly journals Clonal analysis of a human antibody response. Quantitation of precursors of antibody-producing cells and generation and characterization of monoclonal IgM, IgG, and IgA to rabies virus.

1990 ◽  
Vol 171 (1) ◽  
pp. 19-34 ◽  
Author(s):  
Y Ueki ◽  
I S Goldfarb ◽  
N Harindranath ◽  
M Gore ◽  
H Koprowski ◽  
...  
Keyword(s):  
B Cells ◽  
Antibody Response ◽  
B Cell ◽  
Rabies Virus ◽  
Gene Segment ◽  
Human Antibody ◽  
Cell Repertoire ◽  
High Affinity ◽  
Clonal Level ◽  
Vh Gene

We quantitated and characterized the changes in the human B cell repertoire, at the clonal level, before and after immunization with rabies virus. Moreover, we generated 10 monoclonal cell lines producing IgM, IgG, and IgA antibodies to the virus. We found that in healthy subjects, not previously exposed to the virus, nearly 2% of the circulating B lymphocytes were committed to the production of antibodies that bound the virus. These B cells expressed the surface CD5 molecule. The antibodies they produced were polyreactive IgM that displayed a relatively low affinity for the virus components (Kd, 1.0-2.4 x 10(-6) g/microliters). After immunization, different anti-virus (IgG and IgA) antibody-producing cells consistently appeared in the circulation and increased from less than 0.005% to greater than 10% of the total B cells committed to the production of IgG and IgA, respectively. Most of such B cells do not express CD5 and produce monoreactive antibodies of high affinity for rabies virus (Kd, 6.5 x 10(-9) to 1.2 x 10(-10) g/microliters). One of these IgG mAbs efficiently neutralized rabies virus in vitro and in vivo, as detailed elsewhere (Dietzschold, B., P. Casali, Y. Ueki, M. Gore, C. E. Rupprecht, A. L. Notkins, and H. Koprowski, manuscript submitted for publication). Hybridization experiments using probes specific for the different human V gene segment families revealed that cell precursors producing low affinity IgM binding to rabies virus utilized a restricted number of VH gene segments (i.e., only members of the VHIIIb subfamily), whereas cell precursors producing high affinity IgG and IgA to rabies virus utilized an assortment of different VH gene segments (i.e., members of the VHI, VHIII, VHIV, and VHVI families and VHIIIb subfamily). In conclusion, our studies show that EBV transformation in conjunction with limiting dilution technology and somatic cell hybridization techniques are useful methods for quantitating, at the B cell clonal level, the human antibody response to foreign Ags and for generating human mAbs of predetermined specificity and high affinity.

scholarly journals An Activation-Induced Cytidine Deaminase-Independent Mechanism of Secondary VH Gene Rearrangement in Preimmune Human B Cells

2008 ◽  
Vol 181 (11) ◽  
pp. 7825-7834 ◽  
Author(s):  
Nancy S. Longo ◽  
Gabrielle J. Grundy ◽  
Jisoo Lee ◽  
Martin Gellert ◽  
Peter E. Lipsky
Keyword(s):  
B Cells ◽  
Gene Rearrangement ◽  
Cytidine Deaminase ◽  
Human B Cells ◽  
Activation Induced Cytidine Deaminase ◽  
Independent Mechanism ◽  
Vh Gene

scholarly journals Differential usage of an Ig heavy chain variable region gene by human B- cell tumors

Blood ◽  
1993 ◽  
Vol 82 (1) ◽  
pp. 224-230 ◽  
Author(s):  
FK Stevenson ◽  
MB Spellerberg ◽  
J Treasure ◽  
CJ Chapman ◽  
LE Silberstein ◽  
...  
Keyword(s):  
Amino Acid ◽  
B Cells ◽  
B Cell ◽  
Gene Segment ◽  
Variable Region ◽  
Amino Acid Substitutions ◽  
Region Gene ◽  
Heavy Chains ◽  
Vh Gene ◽  
Histologic Types

A monoclonal anti-idiotypic antibody has been raised that recognizes Igs with heavy chains encoded by a member of the VH4 family, the VH4–21 gene segment. The idiotope (Id) is detectable on a high percentage of early B cells in fetal spleen, and is expressed by certain autoantibodies, particularly cold-reactive anti-red blood cell antibodies. Therefore, it was of interest to investigate usage of this VH gene by neoplastic B cells; 81 chronic lymphocytic leukemias (CLLs) involving CD5+ B cells and 62 B-cell lymphomas of varying histologic type have been analyzed. The Id was expressed by only 3 of 81 (3.7%) of the CLLs, indicating a relatively low usage by these tumors. In contrast, the Id was expressed by 9 of 62 (14.5%) of the lymphomas across a range of histologic types, indicating a differential use of the VH4–21 gene among B-cell neoplasms. For three of the Id-positive lymphomas, each of a different histologic class, the nucleotide sequence of the tumor-derived VH gene was determined; the VH4–21 gene was identified, as expected. The sequence from the CLL was identical to the germline sequence, and the marginal zone lymphoma showed only 3 nucleotide changes, 2 of which gave rise to amino acid substitutions. In contrast, the sequence from the follicular lymphoma showed 29 nucleotide changes giving rise to 14 amino acid substitutions, which were scattered among the CDR and FW regions.

scholarly journals The Distal VH Gene Cluster of the Igh Locus Contains Distinct Regulatory Elements with Pax5 Transcription Factor-Dependent Activity in Pro-B Cells

Immunity ◽  
2011 ◽  
Vol 34 (2) ◽  
pp. 175-187 ◽  
Author(s):  
Anja Ebert ◽  
Shane McManus ◽  
Hiromi Tagoh ◽  
Jasna Medvedovic ◽  
Giorgia Salvagiotto ◽  
...  
Keyword(s):  
Transcription Factor ◽  
B Cells ◽  
Gene Cluster ◽  
Regulatory Elements ◽  
Igh Locus ◽  
Dependent Activity ◽  
Vh Gene

scholarly journals The T-Cell Receptor β Variable Gene Promoter Is Required for Efficient Vβ Rearrangement but Not Allelic Exclusion

2004 ◽  
Vol 24 (16) ◽  
pp. 7015-7023 ◽  
Author(s):  
Chun Jeih Ryu ◽  
Brian B. Haines ◽  
Hye Ran Lee ◽  
Yun Hee Kang ◽  
Dobrin D. Draganov ◽  
...  
Keyword(s):  
T Cell ◽  
T Cell Receptor ◽  
Dna Sequences ◽  
Signal Sequence ◽  
Gene Segment ◽  
Gene Promoter ◽  
Simian Virus 40 ◽  
Cell Receptor ◽  
Allelic Exclusion ◽  
Variable Gene

ABSTRACT To investigate the role of promoters in regulating variable gene rearrangement and allelic exclusion, we constructed mutant mice in which a 1.2-kb region of the Vβ13 promoter was either deleted (P13−/−) or replaced with the simian virus 40 minimal promoter plus five copies of Gal4 DNA sequences (P13R/R). In P13−/− mice, cleavage, rearrangement, and transcription of Vβ13, but not the flanking Vβ gene segments, were significantly inhibited. In P13R/R mice, inhibition of Vβ13 rearrangement was less severe and was not associated with any apparent reduction in Vβ13 cleavage. Expression of a T-cell receptor (TCR) transgene blocked cleavages at the normal Vβ13-recombination signal sequence junction and Vβ13 coding joint formation of both wild-type and mutant Vβ13 alleles. However, a low level of aberrant Vβ13 cleavage was consistently detected, especially in TCR transgenic P13R/R mice. These findings suggest that the variable gene promoter is required for promoting local recombination accessibility of the associated Vβ gene segment. Although the promoter is dispensable for allelic exclusion, it appears to suppress aberrant Vβ cleavages during allelic exclusion.

scholarly journals Immunoglobulin gene rearrangements in normal mouse B cells.

1982 ◽  
Vol 2 (7) ◽  
pp. 829-836 ◽  
Author(s):  
P Early ◽  
C Nottenburg ◽  
I Weissman ◽  
L Hood
Keyword(s):  
B Cells ◽  
Heavy Chain ◽  
Germ Line ◽  
Gene Segment ◽  
Immunoglobulin M ◽  
Chain Gene ◽  
Allelic Exclusion ◽  
Immunoglobulin Gene ◽  
Spleen Cells ◽  
Surface Immunoglobulin

We have analyzed the structure of rearranged mu heavy-chain genes obtained from the genomic DNA of normal BALB/c mouse spleen cells expressing surface immunoglobulin M. Examples were found of two types of nonproductive rearrangements, which may be responsible for allelic exclusion in normal B cells. In one of these rearrangements, a germ line D gene segment has joined to the JH4 gene segment but no V/D joining has occurred. We present evidence that D gene segments lie as a cluster between V and J gene segments in the germ line. A comparison of conserved sequences in V and D gene segments suggests that the D gene segments, which are found only in the heavy-chain gene family, may have evolved from V gene segments similar to the Vk family.

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