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The Journal of Immunology, 2001, 167: 2151-2156.
Copyright © 2001 by The American Association of Immunologists

Circulating Human B Cells That Express Surrogate Light Chains Display a Unique Antibody Repertoire1

Eric Meffre*,{dagger}, Michael Chiorazzi* and Michel C. Nussenzweig2,*,{dagger}

* Laboratory of Molecular Immunology, and {dagger} Howard Hughes Medical Institute, The Rockefeller University, New York, NY 10021


   Abstract
Top
 Abstract
Introduction
 Materials and Methods
 Results
 Discussion
 References
 
Circulating human B cells that coexpress V-preB and conventional L chains (V-preB+L+ B cells) are a recently described subset of B cells that express Abs with features of self-reactivity. Initial analysis of V-preB+L+ B cells was limited to Ig-{kappa} and to the small, underused VH5 family. To determine whether Abs commonly expressed by V-preB+L+ B cells show similar features, we analyzed Ig H chains from three highly expressed VH families, VH1, VH3, and VH4, and Ig-{lambda}. We find that VH1 and VH3 Abs expressed by V-preB+L+ B cells resemble VH5 in that they display increased JH6 use, long CDR3s, and an increased frequency of D-D fusions. Abs in all three of these VH families also show skewed D reading frame use resulting in predominance of hydrophobic amino acids, which are counterselected in conventional B cells. Like Ig-{kappa} genes, the Ig-{lambda} genes in V-preB+L+ B cells show long CDR3s, but they differ from Ig-{kappa} genes in that they display no evidence of receptor editing. We conclude that a large number of H and L chain Abs expressed by V-preB+L+ B cells display features associated with self-reactive Abs.


   Introduction
Top
 Abstract
 Introduction
Materials and Methods
 Results
 Discussion
 References
 
Immune self-tolerance is maintained by a combination of central and peripheral mechanisms (reviewed in Refs. 1, 2, 3, 4). Although T cells make a major contribution to the regulation of anti-self Ab production, there are three T cell-independent mechanisms by which self-reactive B cells are silenced. Deletion, anergy, and receptor editing have been studied extensively in mice that carry self-reactive Ab transgenes (reviewed in Refs. 1, 2, 3, 4). In these transgenic models, central deletion of self-reactive B cells is induced by strong B cell receptor (BCR)3 cross-linking, whereas moderate levels of cross-linking induce anergy (5, 6, 7, 8, 9). Anergic B cells complete B cell development and enter the peripheral circulation but are short-lived cells, and under normal circumstances, these cells fail to contribute to the Ab repertoire (10, 11, 12). In contrast, receptor editing replaces autoreactive BCRs by V(D)J recombination, thereby producing novel non-self-reactive Abs (13, 14, 15). Thus, editing differs from the other B cell tolerance mechanisms in that this mechanism spares the self-reactive B cell.

In mice, receptor editing makes an important contribution to the establishment of the normal Ab repertoire (16), but little is known about the relative contributions of deletion, anergy, and editing to tolerance in humans.

V-preB+L+ B cells are a recently described subpopulation of normal human B cells found in the circulation and tonsils, and they accumulate in the joints of some patients with rheumatoid arthritis (17, 18). These cells differ from other peripheral B cells in that in addition to V-preB they also express low levels of recombinase-activating gene (RAG) mRNA. Although the initial analysis was limited to Ig-{kappa} (L) chain and the VH5 gene family, which makes a small contribution to the normal repertoire (19, 20), it showed an unusual Ab repertoire consistent with self-reactivity and Ig-{kappa} receptor editing (18). Based on these observations, it was proposed that V-preB+L+ B cells may represent B cells that have been tolerized (18). To further examine how tolerance might be established in developing B cells in the human, we extended the characterization of the Ab repertoire in V-preB+L+ B cells to VH1, VH3, VH4, and Ig-{lambda}2.


   Materials and Methods
Top
 Abstract
 Introduction
 Materials and Methods
Results
 Discussion
 References
 
Patient samples and cell preparation

V-preB+L+B cells were obtained from three nonrelated healthy donors by a combination of MACS and cell sorting with anti-V-preB mAbs (donors 1–3; Ref. 18). The anti-V-preB Ab was produced against a region of V-preB that shows no Ig homology, and extensive physical measurements failed to detect such cross-reactivity with Ig L or H chains (21). In brief, V-preB+L+ B cells were enriched from PBMCs by negative selection against non-B cells followed by positive selection with anti-VpreB-labeled with PE (22) and MACS anti-PE microbeads (Miltenyi Biotec, Aurburn, CA). Conventional V-preB-L+ B cells were B cell-enriched PBMC that did not bind to the MACS column. After the initial enrichment by negative and positive selection, the cells were stained with FITC human anti-{kappa} and anti-{lambda}, and allophycocyanin anti-CD19 (BD PharMingen, San Diego, CA) and V-preB+L+CD19+ B cells and conventional V-preB-L+CD19+ B cells were purified by sorting on a FACSVantage (BD Biosciences, Mountain View, CA; Ref. 18).

RNA and RT-PCR

Total RNA was extracted from 104–105 purified cells with TRIzol reagent (Life Technologies, Rockville, MD). After DNase I (Boehringer Mannheim, Indianapolis, IN) treatment, RNA was reverse-transcribed in 10 µl with SuperScript II (Life Technologies). For RT-PCR, 1 ml of cDNA was amplified for 35 cycles of 30 s at 94°C, 30 s at 58°C (VH1-Cµ, VH3-Cµ or VH4-Cµ) or at 55°C (V{lambda}-C{lambda}), and 30 s at 72°C with a final 10-min extension at 72°C with HotStarTaq DNA polymerase (Qiagen, Chatsworth, CA) and the following primers: V{lambda}1–8 family consensus sense, 5'-GGG(G/A)TC(T/C)CTGA(C/T/G)CG(A/C/G)TTCTCTGG(C/G)TCC-3'; V{lambda}9 sense, 5'-ATCCCTGATCGCTTCTCAGTCTTG-3'; V{lambda}10 sense, 5'-GATCTCAGAGAGATTATCTGCATCC-3'; C{lambda} antisense, 5'-CACAC(T/C)AGTGTGGCCTTGTTGGCTTG-3'. Sense FR1 VH1, VH3, and VH4 and antisense Cµ primers were as described previously (17, 23). RT-PCR products were analyzed on 2% agarose gels.

Cloning and sequencing

PCR products were gel purified (QIAquick; Qiagen) and cloned into TA vectors (Invitrogen, San Diego, CA). The dsDNA sequences were obtained with antisense Cµ or C{lambda} primers and dye terminator cycle sequencing (PE Applied Biosystems, Foster City, CA). Sequences were analyzed by comparison with Ig BLAST. When two or more identical sequences were found, they were counted as a single clone. IgH CDR3 length was determined by counting amino acid residues between positions 94 and 102 (conserved tryptophan in all JH segments), and D segments were identified following the criteria of Corbett et al. (24). Ig-{lambda} CDR3 length included amino acids between conserved cysteine 88 and the phenylalanine residue embedded in all J{lambda}s (25). Differences in gene distribution between V-preB+L+ B cells and conventional B cells were analyzed with {chi}2 tests, and they were considered significant when p <= 0.05. Student’s t test was used for CDR3-length analysis.


   Results
Top
 Abstract
 Introduction
 Materials and Methods
 Results
Discussion
 References
 
Distinct VH use in V-preB+L+ B cells

To extend the analysis of the Ig H chain repertoire of V-preB+L+ B cells, we cloned and sequenced VH1, VH3, and VH4 genes from V-preB+L+ and conventional B cells obtained from peripheral blood of three healthy donors. The distribution of the VH sequences from each of the three donors was similar, and they are presented together for simplicity. We chose the VH1, VH3, and VH4 families because these genes are well characterized and account for most of the peripheral B cell repertoire (19, 20, 26, 27, 28, 29).

The VH1, VH3, and VH4 repertoire from conventional B cells was similar to that reported by others (Fig. 1Go; Refs. 20, 26, 27, 28, 29). The VH repertoire of VH1 and VH 4 Abs expressed by V-preB+L+ B cells differed from conventional B cells. Half of the VH1 genes were differentially expressed in V-preB+L+ and conventional B cells. VH1–18 and -46 gene use was increased from 11% and 5%, respectively, in conventional B cells to 29% and 16% in V-preB+L+ B cells (Fig. 1Go, top), whereas VH1–2 and -69 gene use was decreased in V-preB+L+ B cells (Fig. 1Go, top). Among the VH4 genes, VH4–34, was counterselected in V-preB+L+ B cells to 4.5% (Fig. 1Go, bottom) compared with 15% in conventional B cells (n = 117; Ref. 30). In contrast to the VH1 and VH4 genes, we found no statistically significant differences in the VH3 repertoire between V-preB+L+ and conventional B cells. However, the VH3 family is a very large family, and therefore, it is difficult to analyze the contribution of individual family members to the repertoire (31). We conclude that the VH1 and VH4 repertoire of V-preB+L+ B cells differs from conventional B cells.



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  		FIGURE 1. VH1, VH3, and VH4 repertoire analysis for conventional and V-preB+L+ B cells. VH1 (top), VH3 (middle), and VH4 (bottom) gene use was determined in conventional (V-preB-L+) and V-preB+L+ B cells from three unrelated normal donors. Conventional (112, 125, and 117) and V-preB+L+ VHDJH (109, 117, and 107) sequences were analyzed for VH1, VH3, and VH4, respectively. *, Statistically significant difference (VH1–2, p = 0.018; VH1–18, p = 0.003; VH1–46, p = 0.025; VH1–69, p = 0.011; and VH4–34, p = 0.02).

 
H chain CDR3s in V-preB+L+ B cells

VH5 genes expressed in V-preB+L+ B cells show long CDR3s with increased JH6 use and frequent diversity gene segment (D) fusions. VH1 and VH3 genes expressed in V-preB+L+ B cells resembled VH5 in that they displayed increased JH6 use when compared with conventional B cells (Fig. 2GoA). Increased JH6 use was particularly favored by VH1–69 genes (62.5% in V-preB+L+ B cells vs 27.9% in conventional B cells; p < 0.001). JH6 is the longest of the human JHs, and consistent with its overuse, V-preB+L+ B cells have longer VH1 IgH CDR3s than conventional B cells (Fig. 2GoB). In addition, D-D fusions were found in 4% of VH1 and VH3 Abs expressed by V-preB+L+ B cells, whereas only 1 of 351 such fusions were identified in the VH1 and VH3 Abs cloned from conventional B cells (4 of 109 VH1 and 4 of 117 VH3; p = 0.006). In contrast to VH1 and VH3, VH4 Abs from V-preB+L+ B cells showed no bias to JH6, normal CDR3s, and no D-D fusions (Fig. 2Go, A and B, and data not shown). We conclude that VH1, VH3 and VH5 genes expressed by V-preB+L+ B cells show increased JH6 use, long CDR3s, and D-D rearrangements, whereas VH4 genes exhibit none of these features.



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  		FIGURE 2. Characteristics of VH genes expressed by conventional and V-preB+L+ B cells. A, JH use in VH1, VH3, and VH4 genes in conventional and V-preB+L+ B cells. Percentage of JH use is indicated. *, Statistically significant difference (VH1, p = 0.02; and VH3, p = 0.04). B, VH CDR3 length in VH1, VH3, and VH4 rearrangements from circulating conventional and V-preB+L+ B cells (VH1, p = 0.0001). CDR3 length in amino acids (aa) is indicated.

 
Specific D gene segment use in V-preB+L+ B cells

Human Abs show preferential use of certain D segments (24). Therefore, we analyzed D region use in V-preB+L+ B cells. We found that D use in VH1 and VH3 Abs in V-preB+L+ B cells differs from D use in conventional B cells (Fig. 3GoA). VH1 and VH3 genes were pooled for this analysis because they displayed similar features. Several of these differences reached statistical significance, including D2–2 and D6–13, which were overrepresented in V-preB+L+ B cells, and D4–23 use, which was underrepresented (Fig. 3GoA). In contrast, D segment gene use was not significantly altered in most VH4 Abs in V-preB+L+ B cells (Fig. 3GoB).



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  		FIGURE 3. D segment use in conventional and V-preB+L+ B cells. A, D segment use in VH1 plus VH3 genes from conventional and V-preB+L+ B cells. VH1 and VH3 genes were pooled for this analysis because they displayed identical features. B, D segment use in VH4 genes from conventional and V-preB+L+ B cells. *, Statistically significant difference (VH1 plus VH3: D2–2, p = 0.03; D6–13, p = 0.04; and D4–23, p = 0.03; and VH4: D3–16, p = 0.04).

 
Most Ds are in reading frame (RF) 2, which encodes hydrophilic amino acids. One-third of all Ds are in RF3, which frequently encodes hydrophobic amino acids, and few Ds are in RF1, which often contains stop codons (24). To determine whether D segment RF use differs between V-preB+L+ and conventional B cells, we compared D segment sequences for VH1, VH3, VH4, and VH5. We find that the incidence of D2–2, D2–15, D6–13 and D4–17 using hydrophobic RF3s was increased in VH1, VH3, and VH5 rearrangements in V-preB+L+ B cells (Fig. 4GoA and data not shown). D3–10 is unusual in that RF1 is hydrophobic, and D3–10 RF1 use increased from 11% in conventional B cells to 35.5% in V-preB+L+ B cells (Ref. 24 and Fig. 4GoB). When considered together, the number of Ds in hydrophobic RFs increased from 33.9% in conventional B cells to 58.2% in V-preB+L+ B cells (Ref. 24 and Fig. 4GoB; p = 0.01). Although Ds with RFs that encode charged amino acids are rare, VH1, VH3, and VH5 Abs with these features were counterselected in V-preB+L+ B cells (D3–3 (RF1) and D3–10 (RF3) (5 and 11% compared with 18 and 36% in conventional B cells; Fig. 4GoB). By contrast, D segment RF use in VH4 Abs in V-preB+L+ B cells is similar to conventional B cells (data not shown). We conclude that VH1, VH3, and VH5 Abs in V-preB+L+ B cells favor specific Ds in hydrophobic RFs that are normally counterselected in conventional B cells.



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  		FIGURE 4. D reading frame use in VH1 plus VH3 genes from circulating conventional and V-preB+L+ B cells. RF use was assigned according to Corbet et al (24 ). A, D2–2, 6–13, 2–15, and 4–17 RF use in VH1 plus VH3 genes from conventional and V-preB+L+ B cells (V-preB+L+, n = 67; conventional B cells, n = 62; p = 0.01). B, D3–10 and 3–3 RF use in VH1 plus VH3 genes from conventional and V-preB+L+ B cells.

 
Ig-{lambda} expression in V-preB+L+ B cells

Ig-{kappa} genes expressed by V-preB+L+ B cells show long CDR3s and are skewed toward upstream V{kappa}s and downstream J{kappa}s consistent with increased secondary V(D)J recombination (18). By analogy to the Ig-{kappa} locus, the human Ig-{lambda} locus should also allow deletional replacement of VJ{lambda}s by receptor editing (32) and would bias the repertoire to more downstream J{lambda}s and upstream V{lambda}s.

To determine whether V-preB+L+ B cells show evidence of Ig-{lambda} receptor editing, we cloned and sequenced the Ig-{lambda} genes expressed by these cells. V-preB+L+ B cells showed no increase in downstream J{lambda} use when compared with conventional B cells; in fact, there was a small increase in upstream J{lambda}1 segment use (Fig. 5GoA; Refs. 33 and 34). In addition, there was no bias for distal V{lambda} segment use (35, 36 ; Fig. 5GoB). We conclude that Ig-{lambda} repertoire in V-preB+L+ B cells shows no skewing to either upstream V{lambda}s or downstream J{lambda}s and therefore no evidence of VJ{lambda} receptor editing.



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  		FIGURE 5. The {lambda} (L) chain repertoire in conventional and V-preB+L+ B cells. A, J{lambda} use in conventional and V-preB+L+ B cells from a pool of three unrelated normal donors. V-preB-L+ (125) and V-preB+L+ V-J{lambda} (129) sequences were analyzed. Percentage of J{lambda} use is indicated. *, J{lambda}1 use was increased in V-preB+L+ B cells (p = 0.04). B, V{lambda} use in J{lambda}3 conventional and V-preB+L+ B cells. The V{lambda} locus is shown, and the percentage of representation of each V{lambda} family is indicated on the y-axis. C, V{lambda} family use in conventional and V-preB+L+ B cells. The percentage of representation of each V{lambda} family is indicated. *, Statistically significant difference (V{lambda}1, p = 0.01).

 
V{lambda} gene family analysis in V-preB+L+ B cells revealed a bias to V{lambda}1 gene use, which increased from 24.4% in conventional B cells to 40.3% in V-preB+L+ B cells (Fig. 5GoC). This increase resulted from the specific amplification of V{lambda}1–36, 44, and 51 in V-preB+L+ B cells in all three normal donors, whereas V{lambda}1–40 and 47 gene use remained unchanged (V{lambda}1–36/44; p = 0.04, data not shown).

Ig-{lambda} genes expressed in V-preB+L+ B cells resemble the Ig-{kappa} genes in that they show an increase in CDR3 length (Fig. 6Go). The 11-aa-long Ig-{lambda} CDR3s increased from 35% in conventional B cells to 50% in V-preB+L+ B cells, whereas short Ig-{lambda} CDR3s (9 aa) decreased from 24% in conventional to 9% in V-preB+L+ B cells (Fig. 6Go). We conclude that the Ig-{lambda} repertoire in V-preB+L+ B cells is skewed to long CDR3s and selected subset of V{lambda}1 genes.



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  		FIGURE 6. Ig-{lambda} CDR3 length in circulating conventional and V-preB+L+ B cells. CDR3 length in amino acids is indicated. *, Statistically significant difference (9 aa, p = 0.002; 11 aa, p = 0.02).

 

   Discussion
Top
 Abstract
 Introduction
 Materials and Methods
 Results
 Discussion
References
 
V-preB+L+ B cells are a small subset of human B cells found in the blood and tonsils of normal individuals. This subset of B cells is enriched in the joints of some patients with rheumatoid arthritis (18). These cells are unusual in that they continue to express genes that are normally turned off during early B cell development and because they express VH5 and Ig-{kappa} genes with long CDR3s reminiscent of anti-self Abs (37, 38, 39, 40).

In this report, we have extended the analysis of the Ig repertoire of V-preB+L+ B cells to the VH1, VH3 and VH4 gene families and Ig-{lambda}. VH1, VH3, and VH4 genes account for most normal human Abs, and Ig-{lambda} is expressed by 30–40% of human B cells (19, 20, 26, 27, 28, 41). The VH1 and VH3 repertoire expressed by V-preB+L+ B cells is similar to VH5 in that it shows an increase in JH6 use, long CDR3s, and D-D fusions. These Abs also show biased D use and overrepresentation of Ds with hydrophobic amino acids (24, 29). In addition, Ig-{lambda} genes with long CDR3s are enriched in V-preB+L+ B cells. Long IgH, Ig-{kappa}, and Ig-{lambda} CDR3s and IgH D-D fusions are normally counterselected in conventional B cells in humans and mice (24, 29, 37). However, these features are prevalent in autoantibodies (37, 40, 42, 43, 44). We conclude that the Ab repertoire in V-preB+L+ B cells frequently shows features consistent with autoreactivity.

Chronic lymphocytic leukemia B cells (B-CLL) frequently express autoantibodies (42, 45, 46). In particular, VH1–69 genes are highly overrepresented in the Abs expressed in B-CLL cells (20% of all cases; Refs. 47, 48). Although VH1–69 gene use was decreased in V-preB+L+ B cells relative to controls, VH1–69 remains one of the two most-used VH1 genes in these cells, representing over 20% of all VH1 genes. The VH1–69 Abs in V-preB+L+ B cells resemble those found in B-CLL cells because B-CLL VH1–69 genes display long CDR3s, biased JH6 use, and a preference for D2–2 and D3–3 use. Therefore, these data suggest that V-preB+L+ B cells may be precursors of B-CLL (23, 49).

Why is the VH4 repertoire in V-preB+L+ B cells different from VH1, VH3, and VH5? It has been suggested that VH4 genes may be particularly prone to producing self-reactive Abs (50, 51, 52). In particular, Abs with VH4–34 variable regions show intrinsic self-reactivity and recognize i/I carbohydrate self-determinants displayed on red blood cells and other cell types (51, 53, 54). It has been shown that germline-encoded VH4–34 gene has intrinsic self-reactivity and that the CDR3 in VH4–34 Abs modulates the affinity for self (55, 56). We would propose that this intrinsic self-reactivity combined with long hydrophobic CDR3s (which themselves have low levels of self-reactivity; Refs. 37, 38, 39) might form Abs with high affinity for self that are deleted centrally. Consistent with this idea, VH4–34 was specifically depleted from the VH4 repertoire of V-preB+L+ B cells.

In addition to their unusual Ab repertoire, V-preB+L+ B cells also showed evidence of Ig-{kappa} receptor editing (18), but we found no evidence for Ig-{lambda} receptor editing in V-preB+L+ B cells. Secondary rearrangements on the Ig-{lambda} locus are theoretically possible, and have been reported in patients with systemic lupus erythematosus (57) and in some transformed cell lines (58, 59). However, other cell lines that edit Ig-{kappa} fail to rearrange Ig-{lambda} (60), and there is no evidence that secondary Ig-{lambda} rearrangements occur under physiologic circumstances. Ig-{kappa} editing requires persistent high levels of RAG expression during a 2-h arrest at the pre-B cell stage of development, whereas B cells that proceed beyond the late pre-B cell and early immature B cell compartment appear to be unable to edit (16). Because Ig-{kappa} gene rearrangement precedes Ig-{lambda} recombination, the window for Ig-{lambda} gene editing in B cell development may be very narrow, and secondary Ig-{lambda} recombination may be an exception rather than the rule.

Long CDR3s are selected against because their expression interferes with normal B cell development either by hindering IgH and L pairing or because Abs with these features are self-reactive. Why, then, are these features found in V-preB+L+ B cells? It has been suggested that V-preB+L+ B cells differ from other B cells in that the Abs they express prevent them from completing B cell development. In this model, B cells expressing Abs that are partially self-reactive would not fully extinguish expression of developmentally regulated genes such as RAG and V-preB and would be selected to become V-preB+L+ B cells. An alternative explanation is that improper interaction between H and L chains would result in Abs that simply fail to produce the adequate signals for immature B cell development. Improper BCR assembly might allow V-preB+L+ B cell survival but not be sufficient for RAG and V-preB gene down-regulation. Our analysis of the VH1, VH3, VH4, VH5, Ig-{kappa}, and Ig-{lambda} repertoire of V-preB+L+ B cells is consistent with either the selection model or altered development and suggests that V-preB+L+ B cells might be more prone to becoming B-CLL cells.


   Acknowledgments
 
We thank members of the Nussenzweig laboratory for comments and discussions.


   Footnotes
 
1 This work was supported by grants from the National Institutes of Health (to M.C.N.). M.C.N. is an investigator in the Howard Hughes Medical Institute. Back

2 Address correspondence and reprint requests to Dr. Michel C. Nussenzweig, The Rockefeller University, Howard Hughes Medical Institute, 1230 York Avenue, New York, NY 10021. E-mail address: nussen{at}mail.rockefeller.edu Back

3 Abbreviations used in this paper: BCR, B cell receptor; RAG, recombination activating gene; RF, reading frame; B-CLL, B-type chronic lymphocytic leukemia. Back

Received for publication February 23, 2001. Accepted for publication June 14, 2001.


   References
Top
 Abstract
 Introduction
 Materials and Methods
 Results
 Discussion
 References
 

  1. Radic, M. Z., M. Weigert. 1995. Origins of anti-DNA antibodies and their implications for B-cell tolerance. Ann. NY Acad. Sci. 764:384.[Medline]
  2. Goodnow, C. C.. 1996. Balancing immunity and tolerance: deleting and tuning lymphocyte repertoires. Proc. Natl. Acad. Sci. USA 93:2264.[Abstract/Free Full Text]
  3. Nemazee, D.. 2000. Receptor selection in B and T lymphocytes. Annu. Rev. Immunol. 18:19.[Medline]
  4. Meffre, E., R. Casellas, M. C. Nussenzweig. 2000. Antibody regulation of B cell development. Nat. Immunol. 1:379.[Medline]
  5. Nemazee, D. A., K. Bürki. 1989. Clonal deletion of B lymphocytes in a transgenic mouse bearing anti-MHC class I antibody genes. Nature 337:562.[Medline]
  6. Hartley, S. B., J. Crosbie, R. Brink, A. B. Kantor, A. Basten, C. C. Goodnow. 1991. Elimination from peripheral lymphoid tissues of self-reactive B lymphocytes recognizing membrane-bound antigens. Nature 353:765.[Medline]
  7. Goodnow, C. C., J. Crosbie, S. Adelstein, T. B. Lavoie, S. J. Smith-Gill, R. A. Brink, H. Pritchard-Briscoe, J. S. Wotherspoon, R. H. Loblay, K. Raphael, et al 1988. Altered immunoglobulin expression and functional silencing of self-reactive B lymphocytes in transgenic mice. Nature 334:676.[Medline]
  8. Erikson, J., M. Z. Radic, S. A. Camper, R. R. Hardy, C. Carmack, M. Weigert. 1991. Expression of anti-DNA immunoglobulin transgenes in non-autoimmune mice. Nature 349:331.[Medline]
  9. Chen, C., E. L. Prak, M. Weigert. 1997. Editing disease-associated autoantibodies. Immunity 6:97.[Medline]
  10. Fulcher, D. A., A. Basten. 1994. Reduced life span of anergic self-reactive B cells in a double-transgenic model. J. Exp. Med. 179:125.[Abstract/Free Full Text]
  11. Cyster, J. G., S. B. Hartley, C. C. Goodnow. 1994. Competition for follicular niches excludes self-reactive cells from the recirculating B-cell repertoire. Nature 371:389.[Medline]
  12. Cyster, J. G., C. C. Goodnow. 1995. Antigen-induced exclusion from follicles and anergy are separate and complementary processes that influence peripheral B cell fate. Immunity 3:691.[Medline]
  13. Gay, D., T. Saunders, S. Camper, M. Weigert. 1993. Receptor editing: an approach by autoreactive B Cells to escape tolerance. J. Exp. Med. 177:999.[Abstract/Free Full Text]
  14. Radic, M. Z., J. Erickson, S. Litwin, M. Weigert. 1993. B lymphocytes may escape tolerance by revising their antigen receptors. J. Exp. Med. 177:1165.[Abstract/Free Full Text]
  15. Tiegs, S. L., D. M. Russell, D. Nemazee. 1993. Receptor editing in self-reactive bone marrow B cells. J. Exp. Med. 177:1009.[Abstract/Free Full Text]
  16. Casellas, R., T. A. Shih, M. Kleinewietfeld, J. Rakonjac, D. Nemazee, K. Rajewsky, M. C. Nussenzweig. 2001. Contribution of receptor editing to the antibody repertoire. Science 291:1541.[Abstract/Free Full Text]
  17. Meffre, E., F. Papavasiliou, P. Cohen, O. de Bouteiller, D. Bell, H. Karasuyama, C. Schiff, J. Banchereau, Y. J. Liu, M. C. Nussenzweig. 1998. Antigen receptor engagement turns off the V(D)J recombination machinery in human tonsil B cells. J. Exp. Med. 188:765.[Abstract/Free Full Text]
  18. Meffre, E., E. Davis, C. Schiff, C. Cunningham-Rundles, L. B. Ivashkiv, L. M. Staudt, J. W. Young, M. C. Nussenzweig. 2000. Circulating human B cells that express surrogate light chains and edited receptors. Nat. Immunol. 1:207.[Medline]
  19. Guigou, V., A. M. Cuisinier, C. Tonnelle, D. Moinier, M. Fougereau, F. Fumoux. 1990. Human immunoglobulin VH and VK repertoire revealed by in situ hybridization. Mol Immunol 27:935.[Medline]
  20. Brezinschek, H. P., R. I. Brezinschek, P. E. Lipsky. 1995. Analysis of the heavy chain repertoire of human peripheral B cells using single-cell polymerase chain reaction. J. Immunol. 155:190.[Abstract]
  21. Gauthier, L., B. Lemmers, V. Guelpa-Fonlupt, M. Fougereau, C. Schiff. 1999. µ-Surrogate light chain physicochemical interactions of the human preB cell receptor: implications for VH repertoire selection and cell signaling at the preB cell stage. J. Immunol. 162:41.[Abstract/Free Full Text]
  22. Lemmers, B., L. Gauthier, V. Guelpa-Fonlupt, M. Fougereau, C. Schiff. 1999. The human ({psi}L+µ-) proB complex: cell surface expression and biochemical structure of a putative transducing receptor. Blood 93:4336.[Abstract/Free Full Text]
  23. Fais, F., F. Ghiotto, S. Hashimoto, B. Sellars, A. Valetto, S. L. Allen, P. Schulman, V. P. Vinciguerra, K. Rai, L. Z. Rassenti, et al 1998. Chronic lymphocytic leukemia B cells express restricted sets of mutated and unmutated antigen receptors. J. Clin. Invest. 102:1515.[Medline]
  24. Corbett, S. J., I. M. Tomlinson, E. L. L. Sonnhammer, D. Buck, G. Winter. 1997. 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. J. Mol. Biol. 270:587.[Medline]
  25. Kabat, E. A., T. T. Wu, H. M. Perry, K. S. Gottesman, C. Foeller. 1991. Sequences of proteins of immunological interest 2387. U.S. Department of Health and Human Services, Bethesda.
  26. Pascual, V., J. D. Capra. 1991. Human immunoglobulin heavy-chain variable region genes: organization, polymorphism, and expression. Adv. Immunol. 49:1.[Medline]
  27. Huang, C., A. K. Stewart, R. S. Schwartz, B. D. Stollar. 1992. Immunoglobulin heavy chain gene expression in peripheral blood B lymphocytes. J. Clin. Invest. 89:1331.
  28. Milner, E. C., W. O. Hufnagle, A. M. Glas, I. Suzuki, C. Alexander. 1995. Polymorphism and utilization of human VH genes. Ann. NY Acad. Sci. 764:50.[Medline]
  29. Brezinschek, H. P., S. J. Foster, R. I. Brezinschek, T. Dorner, R. Domiati-Saad, P. E. Lipsky. 1997. Analysis of the human VH gene repertoire: differential effects of selection and somatic hypermutation on human peripheral CD5+/IgM+ and CD5-/IgM+ B cells. J. Clin. Invest. 99:2488.[Medline]
  30. Kraj, P., S. P. Rao, A. M. Glas, R. R. Hardy, E. C. Milner, L. E. Silberstein. 1997. The human heavy chain Ig V region gene repertoire is biased at all stages of B cell ontogeny, including early pre-B cells. J. Immunol. 158:5824.[Abstract]
  31. Matsuda, F., K. Ishii, P. Bourvagnet, K. Kuma, H. Hayashida, T. Miyata, T. Honjo. 1998. The complete nucleotide sequence of the human immunoglobulin heavy chain variable region locus. J. Exp. Med. 188:2151.[Abstract/Free Full Text]
  32. Vasicek, T. J., P. Leder. 1990. Structure and expression of the human immunoglobulin {lambda} genes. J. Exp. Med. 172:609.[Abstract/Free Full Text]
  33. Ignatovich, O., I. M. Tomlinson, P. T. Jones, G. Winter. 1997. The creation of diversity in the human immunoglobulin V{lambda} repertoire. J Mol Biol 268:69.[Medline]
  34. Popov, A. V., X. Zou, J. Xian, I. C. Nicholson, M. Bruggemann. 1999. A human immunoglobulin {lambda} locus is similarly well expressed in mice and humans. J. Exp. Med. 189:1611.[Abstract/Free Full Text]
  35. Frippiat, J. P., S. C. Williams, I. M. Tomlinson, G. P. Cook, D. Cherif, D. Le Paslier, J. E. Collins, I. Dunham, G. Winter, M. P. Lefranc. 1995. Organization of the human immunoglobulin {lambda} light-chain locus on chromosome 22q11.2. Hum. Mol. Genet. 4:983.[Abstract/Free Full Text]
  36. Williams, S. C., J. P. Frippiat, I. M. Tomlinson, O. Ignatovich, M. P. Lefranc, G. Winter. 1996. Sequence and evolution of the human germline V{lambda} repertoire. J. Mol. Biol. 264:220.[Medline]
  37. Klonowski, K. D., L. L. Primiano, M. Monestier. 1999. Atypical VH-D-JH rearrangements in newborn autoimmune MRL mice. J. Immunol. 162:1566.[Abstract/Free Full Text]
  38. Ichiyoshi, Y., P. Casali. 1994. Analysis of the structural correlates for antibody polyreactivity by multiple reassortments of chimeric human immunoglobulin heavy and light chain V segments. J. Exp. Med. 180:885.[Abstract/Free Full Text]
  39. Crouzier, R., T. Martin, J. L. Pasquali. 1995. Heavy chain variable region, light chain variable region, and heavy chain CDR3 influences on the mono- and polyreactivity and on the affinity of human monoclonal rheumatoid factors. J. Immunol. 154:4526.[Abstract]
  40. Jr Bridges, S. L., S. K. Lee, M. L. Johnson, J. C. Lavelle, P. G. Fowler, W. J. Koopman, Jr H. W. Schroeder. 1995. Somatic mutation and CDR3 lengths of immunoglobulin {kappa} light chains expressed in patients with rheumatoid arthritis and in normal individuals. J. Clin. Invest. 96:831.
  41. Hood, L., W. R. Gray, B. G. Sanders, W. J. Dreyer. 1967. Light chain evolution. Cold Spring Harbor Symp. Quant. Biol. 48:133.
  42. Kipps, T. J., D. A. Carson. 1993. Autoantibodies in chronic lymphocytic leukemia and related systemic autoimmune diseases. Blood 81:2475.[Free Full Text]
  43. Lee, S. K., C. H. Song, J. B. Kim, Y. J. Chwae, I. H. Choi, Jr S. L. Bridges, W. J. Koopman, Jr H. W. Schroeder. 1998. Enhanced expression of immunoglobulin {kappa} light chains with unusually long CDR3 regions in patients with rheumatoid arthritis. J. Rheumatol. 25:1067.[Medline]
  44. Jr Bridges, S. L.. 1998. Frequent N addition and clonal relatedness among immunoglobulin {lambda} light chains expressed in rheumatoid arthritis synovia and PBL, and the influence of V {lambda} gene segment utilization on CDR3 length. Mol. Med. 4:525.[Medline]
  45. Sthoeger, Z. M., M. Wakai, D. B. Tse, V. P. Vinciguerra, S. L. Allen, D. R. Budman, S. M. Lichtman, P. Schulman, L. R. Weiselberg, N. Chiorazzi. 1989. Production of autoantibodies by CD5-expressing B lymphocytes from patients with chronic lymphocytic leukemia. J. Exp. Med. 169:255.[Abstract/Free Full Text]
  46. Borche, L., A. Lim, J. L. Binet, G. Dighiero. 1990. Evidence that chronic lymphocytic leukemia B lymphocytes are frequently committed to production of natural autoantibodies. Blood 76:562.[Abstract/Free Full Text]
  47. Kipps, T. J., E. Tomhave, L. F. Pratt, S. Duffy, P. P. Chen, D. A. Carson. 1989. Developmentally restricted immunoglobulin heavy chain variable region gene expressed at high frequency in chronic lymphocytic leukemia. Proc. Natl. Acad. Sci. USA 86:5913.[Abstract/Free Full Text]
  48. Efremov, D. G., M. Ivanovski, N. Siljanovski, G. Pozzato, L. Cevreska, F. Fais, N. Chiorazzi, F. D. Batista, O. R. Burrone. 1996. Restricted immunoglobulin VH region repertoire in chronic lymphocytic leukemia patients with autoimmune hemolytic anemia. Blood 87:3869.[Abstract/Free Full Text]
  49. Widhopf, G. F. II, T. J. Kipps. 2001. Normal B cells express 51p1-encoded Ig heavy chains that are distinct from those expressed by chronic lymphocytic leukemia B cells. J. Immunol. 166:95.[Abstract/Free Full Text]
  50. Sanz, I., P. Kelly, C. Williams, S. Scholl, P. Tucker, J. D. Capra. 1989. The smaller human VH gene families display remarkably little polymorphism. EMBO J. 8:3741.[Medline]
  51. Silberstein, L. E., L. C. Jefferies, J. Goldman, D. Friedman, J. S. Moore, P. C. Nowell, D. Roelcke, W. Pruzanski, J. Roudier, G. J. Silverman. 1991. Variable region gene analysis of pathologic human autoantibodies to the related i and I red blood cell antigens. Blood 78:2372.[Abstract/Free Full Text]
  52. Pascual, V., J. D. Capra. 1992. VH4–21, a human VH gene segment overrepresented in the autoimmune repertoire. Arthritis Rheum. 35:11.[Medline]
  53. Pascual, V., K. Victor, D. Lelsz, M. B. Spellerberg, T. J. Hamblin, K. M. Thompson, I. Randen, J. Natvig, J. D. Capra, F. K. Stevenson. 1991. Nucleotide sequence analysis of the V regions of two IgM cold agglutinins: evidence that the VH4–21 gene segment is responsible for the major cross-reactive idiotype. J. Immunol. 146:4385.[Abstract]
  54. Potter, K. N., Y. Li, V. Pascual, Jr R. C. Williams, L. C. Byres, M. Spellerberg, F. K. Stevenson, J. D. Capra. 1993. Molecular characterization of a cross-reactive idiotope on human immunoglobulins utilizing the VH4–21 gene segment. J. Exp. Med. 178:1419.[Abstract/Free Full Text]
  55. Li, Y., M. B. Spellerberg, F. K. Stevenson, J. D. Capra, K. N. Potter. 1996. The I binding specificity of human VH 4–34 (VH4–21) encoded antibodies is determined by both VH framework region 1 and complementarity determining region 3. J. Mol. Biol. 256:577.[Medline]
  56. Cauerhff, A., B. C. Braden, J. G. Carvalho, R. Aparicio, I. Polikarpov, J. Leoni, F. A. Goldbaum. 2000. Three-dimensional structure of the fab from a human IgM cold agglutinin. J. Immunol. 165:6422.[Abstract/Free Full Text]
  57. Dorner, T., N. L. Farner, P. E. Lipsky. 1999. Ig{lambda} and heavy chain gene usage in early untreated systemic lupus erythematosus suggests intensive B cell stimulation. J. Immunol. 163:1027.[Abstract/Free Full Text]
  58. Berinstein, N., S. Levy, R. Levy. 1989. Activation of an excluded immunoglobulin allele in a human B lymphoma cell line. Science 244:337.[Abstract/Free Full Text]
  59. Stiernholm, N. B., N. L. Berinstein. 1993. Up-regulated recombination-activating gene expression in sIg- variants of a human mature B cell line undergoing secondary Ig{lambda} rearrangements in cell culture. Eur. J. Immunol. 23:1501.[Medline]
  60. Maes, J., Y. Caspi, F. Rougeon, J. Haimovich, M. Goodhardt. 2000. Secondary V(D)J rearrangements and B cell receptor-mediated down- regulation of recombination activating gene-2 expression in a murine B cell line. J. Immunol. 165:703.[Abstract/Free Full Text]



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