Differential expression of VH gene families in peripheral B cell repertoires of newborn or adult immunoglobulin H chain congenic mice.

The pattern of VH gene family expression in the primary B cell repertoire of the mouse is strain dependent. In C57Bl/6 mice, the VH J558 family is expressed by more than 45% of the cells, while the expression of VH 7183, VH Q52, and VH 36-60 families together does not exceed 20%. In BALB/c mice, relative expression of VH J558 is lower than 35%, while the sum of the other three families reaches 25%. To assess which genetic loci control strain-specific VH gene family expression, we studied VH gene family usage in splenic B cell repertoires of different congenic strains of mice. Changes in major histocompatibility complex or immunoglobulin (Ig) K light chain genes did not modify VH gene family expression in adult mice. Differences at the IgH locus, however, modified VH gene family usage. In 1-d-old mice, the strain-specific VH gene family expression pattern is determined by the IgH haplotype. In adult mice, the VH gene family expression pattern of resting B cells is independent of the IgH locus and follows the genetic background of the congenic strain, while it is determined by the IgH haplotype among Ig-secreting spleen cells. In F1(B6 x BALB/c) mice, each of the two spleen B cell populations, sorted on the basis of mu heavy chain allotype expression, shows an independent VH gene family expression pattern, determined by the IgH locus. The implications of these results in the control of VH gene family expression, and in the selection of peripheral B cell repertoires are discussed.

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Comparison of the fetal and adult functional B cell repertoires by analysis of VH gene family expression.

Journal of Experimental Medicine ◽

10.1084/jem.168.2.589 ◽

1988 ◽

Vol 168 (2) ◽

pp. 589-603 ◽

Cited By ~ 71

Author(s):

H D Jeong ◽

J M Teale

Keyword(s):

Gene Expression ◽

In Situ Hybridization ◽

B Cells ◽

Gene Family ◽

B Cell ◽

Cell Lineage ◽

Cell Repertoire ◽

B Cell Repertoire ◽

Vh Gene

The functional B cell repertoire in BALB/c mice was assessed at various stages in ontogeny. This was done by analyzing VH gene family expression using the sensitive technique of in situ hybridization. The B cell repertoire was probed with the mitogen, LPS, and the antigen DNP. DNP was chosen because B cells responsive to this hapten appear very early in ontogeny. The APCs that developed after stimulation with LPS or DNP were analyzed for VH gene expression by in situ hybridization of individual cells using radiolabeled VH gene family probes. The results indicated that VH gene expression in fetal B cells after stimulation was distinct from adult B cells in that there was a biased expression of D proximal families. The results indicated that this bias was associated with developmental age and not a given differentiation stage in the B cell lineage. In addition, stimulation of fetal B cells with DNP resulted in a large increase in expression of member(s) of VH 36-60, suggesting that the early appearance of DNP-responsive B cells is not strictly correlated with preferential rearrangement of D proximal families, VH 7183 and VH Q52. However, the results suggested that a large proportion of pre-B cells that preferentially rearrange D proximal families early in ontogeny become part of the functional developing repertoire.

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Contribution of the VH11 gene family to mitogen-responsive B cell repertoire in C57BL/6 mice

European Journal of Immunology ◽

10.1002/eji.1830210344 ◽

1991 ◽

Vol 21 (3) ◽

pp. 827-830 ◽

Cited By ~ 5

Author(s):

Azad Kaushik ◽

Constantin Bona ◽

Luc Reininger ◽

Jean-Claude Jaton ◽

Garnett Kelsoe

Keyword(s):

Gene Family ◽

B Cell ◽

Cell Repertoire ◽

B Cell Repertoire

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

Journal of Experimental Medicine ◽

10.1084/jem.177.2.409 ◽

1993 ◽

Vol 177 (2) ◽

pp. 409-418 ◽

Cited By ~ 63

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.

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IgG-Secreting B-Cell Lymphoplasmocytoid Leukaemia : Molecular Characterization of Igh Rearrangements.

Blood ◽

10.1182/blood.v112.11.1778.1778 ◽

2008 ◽

Vol 112 (11) ◽

pp. 1778-1778

Author(s):

Laurence Lode ◽

Surinder S. Sahota ◽

Soraya Wuilleme ◽

Steven Richebourg ◽

Marion Eveillard ◽

...

Keyword(s):

B Cell ◽

Elevated Serum ◽

Chromosomal Translocation ◽

Memory Cell ◽

Fish Analysis ◽

Cell Repertoire ◽

Igh Locus ◽

B Cell Repertoire ◽

Vh Genes

Abstract Purpose : Elevated serum M component of IgG isotype is typically associated with multiple myeloma (MM). However, our group has previously reported cases with an elevated serum monoclonal IgG and a leukaemic B-cell lymphoplasmocytoid lymphoma (LPL) similar to Waldenstrom’s macroglobulinaemia (WM). The aim of this study was to extend analysis of IgH locus events in a larger series of IgG-secreting LPL. Patients and Methods : We investigated 20 patients with an elevated serum monoclonal IgG (>20g/l) and LPL (IgG-LPL). Morphological classification and immunophenotyping analysis were performed at diagnosis (serum IgG>4 g/l, CD19+ cells >30%, presence of lymphoplasmocytoid cells in blood and/or bone marrow). Histological classification and FISH analysis were performed when possible to further characterize those cases. Analysis of VH genes was carried out from RNA with VHLeader and CH primers. 14 patients were examined for both IgG and IgM transcripts; VH-Cμ and VH-Cg transcripts could be compared in 9 patients. Results : Of 25 Ig VH rearrangement sequences, 23 were functional and expressed in each case. VH3 family members appeared to be over-represented (19/21 patients (90.5%) as compared to 40/71 (56.3%) in normal B-cell repertoire (1). VH3-23 was the most frequently used segment (10/21 patients) and is frequently utilized in normal B-cells. IgG-LPL JH family use resembled the normal B-cell repertoire (predominance of JH4 and JH6 segments). The median CDR3 length was 10 amino acids [5–19]. However, and in contrast with features seen in other leukaemia, there was no evidence of homologous CDR3 motifs. All VH genes revealed highly somatically mutated sequences, with a median mutation rate 8.8% [0.7 – 11.1%] (IMGT database(2)). We compared pre (VH-Cμ) and post-switch (VH-Cg) transcripts, and 4/9 patients had identical clonally-derived sequences, and two 2/9 had divergent sequences. Interpretation and conclusion : This intended study of IgG-LPL reveals consistent features that argue for common origins of IgG-LPL with Waldenstrom’s macroglobulinemia. One feature is extensive somatic mutations in VH genes, suggesting origins from a cell that may have undergone successive rounds of mutation. Patterns of mutations in pre-and post-switched clonally derived sequences suggest that the final neoplastic event has occurred in a IgM+ memory cell undergoing isotype switch. However, no aberrant chromosomal translocation accrue at the IgH locus, as apparent from FISH data. This indicates that, unlike in typical MM, switch activity does not generate 14q32 abnormalities nor that such lesions play a role in the pathogenesis of LPL. Extensive mutations are also seen in VH genes in WM, and there is evidence that WM cells can undergo class switch events in vivo. In this tumor, switching occurs at a low subclonal level in some cells, which in rare cases can lead to the emergence of a dual population of clonally identical IgM and IgG expressing WM tumor cells at a later stage of disease (7). In typical WM, 14q32 abnormalities are also generally not seen. These data suggest that IgG-LPL and WM could be two variants of the same entity, with IgG-LPL exposed to persistent switch stimuli following transformation.

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Peripheral T Cells Select the B-Cell Repertoire in Old Mice

Immunological Reviews ◽

10.1111/j.1600-065x.1989.tb00033.x ◽

1989 ◽

Vol 110 (1) ◽

pp. 173-185 ◽

Cited By ~ 24

Author(s):

Marc E. Weksler ◽

Carlo Russo ◽

Gregory W. Siskind

Keyword(s):

T Cells ◽

B Cell ◽

Cell Repertoire ◽

B Cell Repertoire ◽

Peripheral T Cells ◽

Old Mice

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Expression of V Gene Families During Ontogeny and Establishment of B Cell Repertoire

International Reviews of Immunology ◽

10.3109/08830189209055565 ◽

1992 ◽

Vol 8 (2-3) ◽

pp. 83-94 ◽

Cited By ~ 1

Author(s):

Constantin A. Bona

Keyword(s):

B Cell ◽

Gene Families ◽

Cell Repertoire ◽

B Cell Repertoire ◽

V Gene

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Clonotypic heterogeneity in experimental interstitial nephritis. Restricted specificity of the anti-tubular basement membrane B cell repertoire is associated with a disease-modifying crossreactive idiotype.

Journal of Experimental Medicine ◽

10.1084/jem.167.4.1296 ◽

1988 ◽

Vol 167 (4) ◽

pp. 1296-1312 ◽

Cited By ~ 14

Author(s):

M D Clayman ◽

M J Sun ◽

L Michaud ◽

J Brill-Dashoff ◽

R Riblet ◽

...

Keyword(s):

Basement Membrane ◽

B Cell ◽

Interstitial Nephritis ◽

Gene Families ◽

Brown Norway ◽

Renal Lesion ◽

Tubular Basement Membrane ◽

Cell Repertoire ◽

B Cell Repertoire ◽

Disease Modifying

Experimental anti-tubular basement membrane (anti-TBM) disease is an autoimmune interstitial nephritis elicited in susceptible rodents after immunization with renal tubular antigen. The nephritogenic antigen in the immunizing preparation is 3M-1, a 48,000 Mr noncollagenous glycoprotein. The hallmarks of the renal lesion are the presence of anti-TBM antibodies (anti-TBM-Ab) and a dense mononuclear cell infiltrate. The anti-TBM B cell repertoire in this disease was analyzed using a library of 22 anti-TBM mAbs generated in a prototypically susceptible Brown Norway rat. These anti-TBM mAbs were all demonstrated to be 3M-1 specific and their characterization formed the basis for the following observations: (a) The size of the anti-TBM B cell population is estimated at 58 distinct clones; (b) by competitive inhibition criteria, all anti-TBM mAbs recognize the same (or spatially close) epitope(s) on 3M-1. This focused recognition was maintained in spite of considerable variability in affinity. Epitopic dominance could also be demonstrated in human polyclonal anti-TBM antisera from a patient with anti-TBM disease; and (c) a crossreactive idiotype was documented, and antisera directed toward this set of variable region determinants was shown to be effective as a prophylactic regimen to abrogate disease, and as a therapeutic modality to arrest the progression of disease; (d) analysis of VH gene families suggested biased usage of Q52- and 7183-like families, although at least three gene families are used in the anti-TBM-Ab response. Thus, the anti-TBM B cell compartment in BN rats is moderately large, but is primarily focused to a single epitope on the nephritogenic antigen and is associated with a disease-modifying crossreactive idiotype.

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