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Science 20 December 1996:
Vol. 274. no. 5295, pp. 2094 - 2097
DOI: 10.1126/science.274.5295.2094
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Reports

Neoteny in Lymphocytes: Rag1 and Rag2 Expression in Germinal Center B Cells

Shuhua Han, Biao Zheng, David G. Schatz, Eugenia Spanopoulou, Garnett Kelsoe *

The products of the Rag1 and Rag2 genes drive genomic V(D)J rearrangements that assemble functional immunoglobulin and T cell antigen receptor genes. Expression of the Rag genes has been thought to be limited to developmentally immature lymphocyte populations that in normal adult animals are primarily restricted to the bone marrow and thymus. Abundant RAG1 and RAG2 protein and messenger RNA was detected in the activated B cells that populate murine splenic and Peyer's patch germinal centers. Germinal center B cells thus share fundamental characteristics of immature lymphocytes, raising the possibility that antigen-dependent secondary V(D)J rearrangements modify the peripheral antibody repertoire.

S. Han, B. Zheng, G. Kelsoe, Department of Microbiology and Immunology, University of Maryland School of Medicine, 655 West Baltimore Street, Baltimore, MD 21201, USA.
D. G. Schatz, Section of Immunobiology, Howard Hughes Medical Institute, Yale University School of Medicine, New Haven, CT 06520, USA.
E. Spanopoulou, Mount Sinai Cancer Center, New York, NY 10021, USA.
*   To whom correspondence should be addressed.

Periodic expression of the recombination-activating genes Rag1 and Rag2 controls the assembly of immunoglobulin (Ig) genes and defines the principal stages of B lymphopoiesis in the bone marrow (1). Transcription of the Rag genes ends with the expression of competent Ig on the surface of immature B cells, precluding further V(D)J recombination in the mature lymphocyte pool (2). However, we and others have found that lymphocytes in germinal centers (GCs) exhibit features of immature T and B cells, including the expression of membrane markers typically present on developing lymphocytes (3) and exquisite sensitivity to activation-induced apoptosis that is independent of the Fas molecule (4). Perhaps most remarkable is the similar spectrum of nucleotide exchanges introduced during antigen-driven V(D)J hypermutation in murine GCs and by the developmentally regulated generation of point mutations in the Ig genes of B cells in ileal Peyer's patches (PP) of fetal lambs (5).

Germinal centers are sites of antigen- and T cell-dependent cellular reactions that develop in secondary lymphoid tissues. Germinal centers are necessary for immunological memory in the B cell compartment (6, 7) and are the site of V(D)J hypermutation and selection that is required for affinity maturation of antibody responses (8). Two populations of B lymphocytes, the mitotically active Ig- centroblasts and the nondividing Ig+ centrocytes, make up the majority of GC cells; centrocytes arise from centroblasts, and in turn, some centrocytes reenter the proliferating pool (3, 9). Evidence suggests that the centrocyte population is subject to selective apoptosis (4). Splenic GCs first appear 4 to 5 days after primary immunization and may be identified by their distinctive ability to bind peanut agglutinin (PNA+) and the monoclonal antibody GL-7 (GL-7+) (3). The GC reaction is transient, peaking by day 12 of the response and waning after 3 weeks (9). In contrast, GCs are constitutively present in murine PPs, being chronically stimulated by food antigens and the gut flora (10).

To determine if the immature character of GC B cells extended to the level of Rag expression, we used affinity-purified antibodies specific for active RAG1 and RAG2 proteins (11) to label histologic sections of spleen and PP from immunized and normal mice (12). Mature GCs, those present in spleen 16 days after immunization (Fig. 1) or in the PPs of unimmunized mice (Fig. 2), contain PNA+, GL-7+ B cells that express substantial amounts of immunoreactive RAG1 and RAG2 protein. The distribution of labeled cells coincided with the distribution of B7-2 expression, suggesting that RAG proteins are predominantly expressed in the centrocytes of the GC light zone (7). Virtually identical staining patterns for immunoreactive RAG1 were achieved with rabbit IgG specific for the NH2-terminal residues of RAG1 and a murine monoclonal antibody that binds to the COOH-terminal region of RAG1 (13). Histologic demonstration of RAG2 was more difficult, even in sections of thymus, a site of active V(D)J recombination and high RAG expression (1). Two rabbit antibodies were used to localize RAG2 protein; one, made against amino acids 70 through 516 of murine RAG2, gave equivocal labeling, whereas the other (antibody 435), specific for a 20-amino acid stretch of RAG2 (13), adequately labeled both GCs (Figs. 1C and 2B) and cortical thymocytes.

Fig. 1. Immunohistological staining of a single GC in serial splenic sections. Adjacent, 6-µm sections (A to E) through the spleen of a C57BL/6 mouse immunized with NP-CGG 16 days earlier were stained with (A) peanut agglutinin (PNA) (red), (B) rabbit antibody to RAG1 (anti-RAG1) (blue), (C) rabbit anti-RAG2 (blue), (D) normal rabbit Ig (blue), and (E) GL-7 antibody (blue). (F) The GC structure and adjacent splenic architecture are diagrammatically illustrated. GL-7 and PNA label centroblasts and centrocytes to define the location of the GC through the intervening sections; note that the GC column rotates clockwise as it follows the periarteriolar lymphoid sheath and central arteriole through the splenic white pulp. LZ, light zone; DZ, dark zone; ca, central arteriole; pals, periarteriolar lymphoid sheath, the splenic T cell zone. Magnification ×90. [View Larger Version of this Image (108K GIF file)]

Fig. 2. Expression of RAG1 and RAG2 in PP germinal centers. Adjacent sections of PP from naïve C57BL/6 mice were labeled with (A) rabbit anti-RAG1 (blue) and PNA-HRP (red), (B) anti-RAG2 (blue) and PNA-HRP (red), and (C) normal rabbit Ig (blue) plus PNA-HRP, as described (12). Doubly labeled cells appear black. LZ, light zone; DZ, dark zone; ser, serosal surface of the PP. Magnification ×208. [View Larger Version of this Image (88K GIF file)]

The presence of immunoreactive RAG1 and RAG2 in GC B cells was further supported by using the reverse transcriptase-dependent polymerase chain reaction (RT-PCR) assay to detect the presence of RAG1, RAG2, and hypoxanthine-guanine phosphoribosyl transferase (HPRT) mRNA (14) in small numbers (5 × 103 to 2 × 104 cells) of GC (GL-7+B220+) and follicular (GL-7-B220+) B cells purified by fluorescence-activated cell sorting (15) from spleens of immunized mice. Comparable numbers of immature, CD4+CD8+ (double-positive) thymocytes were similarly prepared to serve as controls for the RT-PCR assay. RAG1 and RAG2 message was readily detected in double-positive thymocytes and in as few as 5 × 103 GC B cells (Fig. 3). Re-analysis of sorted GC B lymphocytes indicated enrichment of GL-7+B220+ cells to only sim 35% compared with >=96% for follicular B cells and double-positive thymocytes. In contrast, neither RAG1 nor RAG2 message could be detected in even larger numbers (2 × 104 cells) of follicular B cells or lipopolysaccharide-activated B cell blasts (Fig. 3). Approximately equivalent amounts of HPRT mRNA were present in all cell cohorts, indicating generally equivalent recoveries of intact RNA (Fig. 3). Sequence analysis of RAG RT-PCR products from GC B cells confirmed these to be RAG1 and RAG2.

Fig. 3. Flow cytometric analysis and RT-PCR assay to evaluate RAG expression in GC B cells. Single-cell suspensions of splenocytes were prepared from three C57BL/6 mice 16 days after immunization with NP-CGG for the isolation of GC and follicular B cells by flow cytometry. (A) Total splenic cells were stained as described (15). Percentages of gated cells for each population are indicated. B220 versus GL-7 defines GC B cells (GL-7+B220+) and follicular B cells (GL-7-B220+). (B to D) Subpopulations including GL-7+B220+ and GL-7-B220+ spleen cells and CD4+CD8+ thymocytes were isolated (range: 5 × 103 to 2 × 104 cells). Cellular RNA was reverse transcribed with primers specific for Rag1, Rag2, and HPRT (14). The cDNA was then amplified with 5' and nested 3' primers (15) specific for (B) Rag1, (C) Rag2, and (D) HPRT, yielding products of 546, 472, and 249 bp, respectively. PCR products were detected by staining with ethidium bromide. Lane 1, molecular size markers; lane 2, no reverse transcriptase control (1 × 104 CD4+CD8+ thymocytes); lane 3, 1 × 104 CD4+CD8+ thymocytes; lane 4, 5 × 103 CD4+CD8+ thymocytes; lane 5, 1 × 104 GL-7+B220+ spleen cells; lane 6, 5 × 103 GL-7+B220+ spleen cells; lane 7, 2 × 104 GL-7-B220+ spleen cells; lane 8, 1 × 104 GL-7-B220+ spleen cells; lane 9, 2 × 104 lipopolysaccharide (LPS)-stimulated (48 hours) spleen cells; lane 10, 1 × 104 LPS-stimulated (48 hours) spleen cells; lane 11, molecular size markers. [View Larger Version of this Image (44K GIF file)]

GC B cells have been proposed to represent a distinct lineage of B lymphocytes (16). Do the B cells that migrate into nascent GCs already carry the RAG proteins or is their expression induced by the GC microenvironment? In contrast to mature GCs, only about half of newly developed splenic GCs contain B cells that express detectable amounts of RAG proteins. The number of RAG+ centrocytes in GCs and the intensity of their labeling increases during the GC reaction, suggesting that events within the GC microenvironment up-regulate RAG1 and RAG2 expression (Fig. 4). This pattern of expression mirrors the onset of µ rightarrow  gamma 1 Ig class switching and the accumulation of point mutations in the Ig heavy chain genes of GC B cells (17). However, Ig class switching simultaneously occurs in RAG- B cells located within extrafollicular foci of antibody-secreting cells (18), and immunization with pneumococcal vaccine, a type-II T cell-independent antigen (19), induces RAG1+ GCs in the absence of significant levels of V(D)J hypermutation (20). These observations imply that RAG proteins are not necessary for Ig class switching nor sufficient for V(D)J hypermutation. Indeed, extensively mutated Ig light chain transgenes have been recovered from Ig transgenic Rag1null mice reconstituted with specific T helper cells and antigen (21). By day 19 after immunization, a fraction of B cells within GCs had lost the ability to bind PNA or GL-7 but remained positive for immunoreactive RAG1 and RAG2. RAG2 protein also persists in newly generated B cells beyond the cessation of transcription (2), suggesting that the V(D)J recombinase may be briefly present in B cells that have exited GCs.

Fig. 4. Kinetics of RAG expression in GCs. C57BL/6 mice were immunized with NP-CGG (12); spleens were removed at days 6, 9, 12, 16, and 19 after immunization and frozen for immunohistology. Sections were co-stained with rabbit anti-RAG1 and PNA. The fraction (percent of total PNA+ GCs; shaded bars) of PNA+ GCs that also expressed immunoreactive RAG1 (>=5% RAG+ cells per GC) were determined by the inspection of 474 PNA+ GCs. The frequencies (percent of total PNA+ cells; black bars) of RAG1+ cells in RAG+ GCs were established by direct enumeration in a randomly selected subset (118 GCs) of this population. Data for days 6 through 16 are means from two to three mice at each time point; the value for day 19 represents a single mouse. [View Larger Version of this Image (39K GIF file)]

Expression of RAG1 and RAG2 in GCs reveals the GC microenvironment as a site that supports a population of peripheral B cells profoundly similar to pre-B cells in the bone marrow. The many phenotypic characters shared by developing B cells and those in GCs (3) extend to reactivation of the V(D)J recombinase. The antigen-dependent GCs of mice may represent evolutionary homologs of gut-associated tissues that drive developmentally regulated diversification of Ig after rearrangement in other vertebrate species (3, 5, 22). However, in mice these properties are present in both the intestinal PPs and splenic GCs (Figs. 1 and 2). Induction of an immature-like state in GC lymphocytes may reflect a mechanism to remove autoreactive cells that arise by mutation (4, 9). The physiologic state that permits this selective apoptosis may coordinately reactivate RAG1 and RAG2.

The availability of V(D)J recombinase in GC B cells also suggests several possibilities for the diversification of Ig genes in centrocytes. RAG1 and RAG2 could mediate secondary V(D)J rearrangements leading to light chain replacement or the introduction of new VH gene segments by means of cryptic recombination signals present near their 3' termini (23). Light chain receptor editing is commonly observed in autoreactive immature B cells driven to initiate apoptosis by Ig engagement--a scenario not unlike the fate of self-reactive centrocytes (4). In fact, a significant fraction of human B cells that express the lambda  light chain carry productively rearranged kappa  light chain genes that have been inactivated by somatic mutation (24). Other evidence consistent with light chain replacement comes from genetic analysis of follicular lymphomas, tumors that exhibit many characteristics of GC lymphocytes including V(D)J hypermutation (25). Sklar et al. (26) have reported one tumor composed of two clonal lymphomas related by a common Ig heavy chain rearrangement but distinct by virtue of dissimilar light chain genes. It may be significant also that the t(14;18) chromosomal translocation present in most follicular lymphomas is thought to arise as an error of V(D)J recombination (27). Documentation of secondary V(D)J rearrangements in GCs would constitute a striking exception to one of immunology's fundamental tenets: that antigen does not elicit the formation of novel receptors.

Note added in proof: Messenger RNA specific for the lambda 5 component of the pre-B cell receptor complex (2) can be readily detected by a specific RT-PCR assay in as few as 5 × 103 GC (GL-7+B220+) B cells. In contrast, lambda 5 message was not found in larger numbers (5 × 104) of follicular B cells (GL-7-B220+). These findings further substantiate the immature character of B lymphocytes in GCs.

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  28. We thank J. Cerny and M. Schlissel for their review of this manuscript; S. V. Desiderio for RAG2-specific antibody; R. Hodes, K. Holmes, and A. Lanzavecchia for describing unpublished observations; and F. W. Alt for the communication of unpublished results confirming Ig class switching in the absence of RAG proteins. Supported in part by USPHS grant AI32524 to D.G.S. and grants AI24335, AG10207, and AG13789 to G.K. E.S. is the recipient of a grant from the Leukemia Research Foundation. D.G.S. is an assistant investigator of the Howard Hughes Medical Institute.

30 July 1996; accepted 4 October 1996


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Ann Rheum Dis 65, 1351-1356
   Abstract »    Full Text »    PDF »
Analysis of RAG expression by peripheral blood CD5+ and CD5- B cells of patients with childhood systemic lupus erythematosus.
H Morbach, S K Singh, C Faber, P E Lipsky, and H J Girschick (2006)
Ann Rheum Dis 65, 482-487
   Abstract »    Full Text »    PDF »
Expression of RAGs in Peripheral B Cells outside Germinal Centers Is Associated with the Expression of CD5.
S. Hillion, A. Saraux, P. Youinou, and C. Jamin (2005)
J. Immunol. 174, 5553-5561
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Receptor editing in peripheral B cell tolerance.
J. S. Rice, J. Newman, C. Wang, D. J. Michael, and B. Diamond (2005)
PNAS 102, 1608-1613
   Abstract »    Full Text »    PDF »
Cutting Edge: TCR Revision Occurs in Germinal Centers.
C. J. Cooper, G. L. Turk, M. Sun, A. G. Farr, and P. J. Fink (2004)
J. Immunol. 173, 6532-6536
   Abstract »    Full Text »    PDF »
Immortalized Dendritic Cell Line with Efficient Cross-Priming Ability Established from Transgenic Mice Harboring the Temperature-Sensitive SV40 Large T-Antigen Gene.
S. Ebihara, S. Endo, K. Ito, Y. Ito, K. Akiyama, M. Obinata, and T. Takai (2004)
J. Biochem. 136, 321-328
   Abstract »    Full Text »    PDF »
CD27 Is Acquired by Primed B Cells at the Centroblast Stage and Promotes Germinal Center Formation.
Y. Xiao, J. Hendriks, P. Langerak, H. Jacobs, and J. Borst (2004)
J. Immunol. 172, 7432-7441
   Abstract »    Full Text »    PDF »
Rectification of age-related impairment in Ig gene hypermutation during a memory response.
S. Han, E. Marinova, and B. Zheng (2004)
Int. Immunol. 16, 525-532
   Abstract »    Full Text »    PDF »
Inflammation Controls B Lymphopoiesis by Regulating Chemokine CXCL12 Expression.
Y. Ueda, K. Yang, S. J. Foster, M. Kondo, and G. Kelsoe (2004)
J. Exp. Med. 199, 47-58
   Abstract »    Full Text »    PDF »
Receptor revision of immunoglobulin heavy chain genes in human MALT lymphomas.
D Lenze, A Greiner, C Knorr, I Anagnostopoulos, H Stein, and M Hummel (2003)
Mol. Pathol. 56, 249-255
   Abstract »    Full Text »    PDF »
T Cell Receptor Revision Does Not Solely Target Recent Thymic Emigrants.
C. J. Cooper, M. T. Orr, C. J. McMahan, and P. J. Fink (2003)
J. Immunol. 171, 226-233
   Abstract »    Full Text »    PDF »
Expression of recombination-activating gene in mature peripheral T cells in Peyer's patch.
E. Kondo, H. Wakao, H. Koseki, T. Takemori, S. Kojo, M. Harada, M. Takahashi, S. Sakata, C. Shimizu, T. Ito, et al. (2003)
Int. Immunol. 15, 393-402
   Abstract »    Full Text »    PDF »
Targeting Apoptotic Tumor Cells to Fc{gamma}R Provides Efficient and Versatile Vaccination Against Tumors by Dendritic Cells.
K. Akiyama, S. Ebihara, A. Yada, K. Matsumura, S. Aiba, T. Nukiwa, and T. Takai (2003)
J. Immunol. 170, 1641-1648
   Abstract »    Full Text »    PDF »
RAG-dependent peripheral T cell receptor diversification in CD8+ T lymphocytes.
P. Serra, A. Amrani, B. Han, J. Yamanouchi, S. J. Thiessen, and P. Santamaria (2002)
PNAS 99, 15566-15571
   Abstract »    Full Text »    PDF »
Chronic Graft-Versus-Host in Ig Knockin Transgenic Mice Abrogates B Cell Tolerance in Anti-Double-Stranded DNA B Cells.
D. R. Sekiguchi, S. M. Jainandunsing, M. L. Fields, M. A. Maldonado, M. P. Madaio, J. Erikson, M. Weigert, and R. A. Eisenberg (2002)
J. Immunol. 168, 4142-4153
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Differential surrogate light chain expression governs B-cell differentiation.
Y.-H. Wang, R. P. Stephan, A. Scheffold, D. Kunkel, H. Karasuyama, A. Radbruch, and M. D. Cooper (2002)
Blood 99, 2459-2467
   Abstract »    Full Text »    PDF »
BCR signal through {alpha}4 is involved in S6 kinase activation and required for B cell maturation including isotype switching and V region somatic hypermutation.
S. Inui, K. Maeda, D. R. Hua, T. Yamashita, H. Yamamoto, E. Miyamoto, S. Aizawa, and N. Sakaguchi (2002)
Int. Immunol. 14, 177-187
   Abstract »    Full Text »    PDF »
Expression of the recombination-activating genes in extrafollicular lymphocytes but no apparent reinduction in germinal center reactions in human tonsils.
N. Meru, A. Jung, I. Baumann, and G. Niedobitek (2002)
Blood 99, 531-537
   Abstract »    Full Text »    PDF »
Identification of Multiple Isolated Lymphoid Follicles on the Antimesenteric Wall of the Mouse Small Intestine.
H. Hamada, T. Hiroi, Y. Nishiyama, H. Takahashi, Y. Masunaga, S. Hachimura, S. Kaminogawa, H. Takahashi-Iwanaga, T. Iwanaga, H. Kiyono, et al. (2002)
J. Immunol. 168, 57-64
   Abstract »    Full Text »    PDF »
Development and Maintenance of a B220- Memory B Cell Compartment.
D. J. Driver, L. J. McHeyzer-Williams, M. Cool, D. B. Stetson, and M. G. McHeyzer-Williams (2001)
J. Immunol. 167, 1393-1405
   Abstract »    Full Text »    PDF »
Localization of recombination activating gene 1/green fluorescent protein (RAG1/GFP) expression in secondary lymphoid organs after immunization with T-dependent antigens in rag1/gfp knockin mice.
H. Igarashi, N. Kuwata, K. Kiyota, K. Sumita, T. Suda, S. Ono, S. R. Bauer, and N. Sakaguchi (2001)
Blood 97, 2680-2687
   Abstract »    Full Text »    PDF »
Novel Secondary Ig VH Gene Rearrangement and In-Frame Ig Heavy Chain Complementarity-Determining Region III Insertion/Deletion Variants in De Novo Follicular Lymphoma.
C. B. Kobrin, M. Bendandi, and L. W. Kwak (2001)
J. Immunol. 166, 2235-2243
   Abstract »    Full Text »    PDF »
Reduction in DNA Binding Activity of the Transcription Factor Pax-5a in B Lymphocytes of Aged Mice.
J. Anspach, G. Poulsen, I. Kaattari, R. Pollock, and P. Zwollo (2001)
J. Immunol. 166, 2617-2626
   Abstract »    Full Text »    PDF »
The Long Isoform of Terminal Deoxynucleotidyl Transferase Enters the Nucleus and, Rather than Catalyzing Nontemplated Nucleotide Addition, Modulates the Catalytic Activity of the Short Isoform.
C. L. Benedict, S. Gilfillan, and J. F. Kearney (2001)
J. Exp. Med. 193, 89-100
   Abstract »    Full Text »    PDF »
RAG1 and RAG2 Expression by B Cell Subsets from Human Tonsil and Peripheral Blood.
H. J. Girschick, A. C. Grammer, T. Nanki, M. Mayo, and P. E. Lipsky (2001)
J. Immunol. 166, 377-386
   Abstract »    Full Text »    PDF »
c-Myb Binds to a Sequence in the Proximal Region of the RAG-2 Promoter and Is Essential for Promoter Activity in T-Lineage Cells.
Q.-F. Wang, J. Lauring, and M. S. Schlissel (2000)
Mol. Cell. Biol. 20, 9203-9211
   Abstract »    Full Text »
Antigen-independent Appearance of Recombination Activating Gene (RAG)-positive Bone Marrow B Cells in the Spleens of Immunized Mice.
F. Gartner, F. W. Alt, R. J. Monroe, and K. J. Seidl (2000)
J. Exp. Med. 192, 1745-1754
   Abstract »    Full Text »    PDF »
Complement C4 Inhibits Systemic Autoimmunity through a Mechanism Independent of Complement Receptors CR1 and CR2.
Z. Chen, S. B. Koralov, and G. Kelsoe (2000)
J. Exp. Med. 192, 1339-1352
   Abstract »    Full Text »    PDF »
Heavy Chain Revision in MRL Mice: A Potential Mechanism for the Development of Autoreactive B Cell Precursors.
K. D. Klonowski and M. Monestier (2000)
J. Immunol. 165, 4487-4493
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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. Maes, Y. Caspi, F. Rougeon, J. Haimovich, and M. Goodhardt (2000)
J. Immunol. 165, 703-709
   Abstract »    Full Text »    PDF »
Immunization and Infection Change the Number of Recombination Activating Gene (RAG)-expressing B Cells in the Periphery by Altering Immature Lymphocyte Production.
H. Nagaoka, G. Gonzalez-Aseguinolaza, M. Tsuji, and M. C. Nussenzweig (2000)
J. Exp. Med. 191, 2113-2120
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Follicular lymphomas' BCL-2/IgH junctions contain templated nucleotide insertions: novel insights into the mechanism of t(14;18) translocation.
U. Jager, S. Bocskor, T. Le, G. Mitterbauer, I. Bolz, A. Chott, M. Kneba, C. Mannhalter, and B. Nadel (2000)
Blood 95, 3520-3529
   Abstract »    Full Text »    PDF »
Revising B Cell Receptors.
D. Nemazee and M. Weigert (2000)
J. Exp. Med. 191, 1813-1818
   Abstract »    Full Text »    PDF »
Receptor Revision of Immunoglobulin Heavy Chain Variable Region Genes in Normal Human B Lymphocytes.
P. C. Wilson, K. Wilson, Y.-J. Liu, J. Banchereau, V. Pascual, and J. D. Capra (2000)
J. Exp. Med. 191, 1881-1894
   Abstract »    Full Text »    PDF »
Humoral Immune Responses in Cr2-/- Mice: Enhanced Affinity Maturation but Impaired Antibody Persistence.
Z. Chen, S. B. Koralov, M. Gendelman, M. C. Carroll, and G. Kelsoe (2000)
J. Immunol. 164, 4522-4532
   Abstract »    Full Text »    PDF »
Cutting Edge: Recombinase-Activating Gene Expression and V(D)J Recombination in CD4+CD3low Mature T Lymphocytes.
E. Lantelme, B. Palermo, L. Granziero, S. Mantovani, R. Campanelli, V. Monafo, A. Lanzavecchia, and C. Giachino (2000)
J. Immunol. 164, 3455-3459
   Abstract »    Full Text »    PDF »
A novel nuclear phosphoprotein, GANP, is up-regulated in centrocytes of the germinal center and associated with MCM3, a protein essential for DNA replication.
K. Kuwahara, M. Yoshida, E. Kondo, A. Sakata, Y. Watanabe, E. Abe, Y. Kouno, S. Tomiyasu, S. Fujimura, T. Tokuhisa, et al. (2000)
Blood 95, 2321-2328
   Abstract »    Full Text »    PDF »
Cutting Edge: Absence of Expression of RAG1 in Peritoneal B-1 Cells Detected by Knocking into RAG1 Locus with Green Fluorescent Protein Gene.
N. Kuwata, H. Igarashi, T. Ohmura, S. Aizawa, and N. Sakaguchi (1999)
J. Immunol. 163, 6355-6359
   Abstract »    Full Text »    PDF »
Regulated Genomic Instability and Neoplasia in the Lymphoid Lineage.
G. J. Vanasse, P. Concannon, and D. M. Willerford (1999)
Blood 94, 3997-4010
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Distinct Signal Thresholds for the Unique Antigen Receptor–linked Gene Expression Programs in Mature and Immature B Cells.
R. J. Benschop, D. Melamed, D. Nemazee, and J. C. Cambier (1999)
J. Exp. Med. 190, 749-756
   Abstract »    Full Text »    PDF »
B Lymphocyte Selection and Age-Related Changes in VH Gene Usage in Mutant Alicia Rabbits.
X. Zhu, A. Boonthum, S.-K. Zhai, and K. L. Knight (1999)
J. Immunol. 163, 3313-3320
   Abstract »    Full Text »    PDF »
Somatic Mutation and Light Chain Rearrangement Generate Autoimmunity in Anti–single-stranded DNA Transgenic MRL/lpr Mice.
F. Brard, M. Shannon, E. L. Prak, S. Litwin, and M. Weigert (1999)
J. Exp. Med. 190, 691-704
   Abstract »    Full Text »    PDF »
An inverse relationship between T cell receptor affinity and antigen dose during CD4+ T cell responses in vivo and in vitro.
W. Rees, J. Bender, T. K. Teague, R. M. Kedl, F. Crawford, P. Marrack, and J. Kappler (1999)
PNAS 96, 9781-9786
   Abstract »    Full Text »    PDF »
Antigen-Induced Somatic Diversification of Rabbit IgH Genes: Gene Conversion and Point Mutation.
C. R. Winstead, S.-K. Zhai, P. Sethupathi, and K. L. Knight (1999)
J. Immunol. 162, 6602-6612
   Abstract »    Full Text »    PDF »
Distinct Factors Regulate the Murine RAG-2 Promoter in B- and T-Cell Lines.
J. Lauring and M. S. Schlissel (1999)
Mol. Cell. Biol. 19, 2601-2612
   Abstract »    Full Text »    PDF »
Gene Conversion and Hypermutation During Diversification of VH Sequences in Developing Splenic Germinal Centers of Immunized Rabbits.
E. Schiaffella, D. Sehgal, A. O. Anderson, and R. G. Mage (1999)
J. Immunol. 162, 3984-3995
   Abstract »    Full Text »    PDF »
Activated Ras Signals Developmental Progression of Recombinase-activating Gene (RAG)-deficient Pro-B Lymphocytes.
A. C. Shaw, W. Swat, R. Ferrini, L. Davidson, and F. W. Alt (1999)
J. Exp. Med. 189, 123-129
   Abstract »    Full Text »    PDF »
RAG Expression in B Cells in Secondary Lymphoid Tissues.
W. YU, H. NAGAOKA, Z. MISULOVIN, E. MEFFRE, H. SUH, M. JANKOVIC, N. YANNOUTSOS, R. CASELLAS, E. BESMER, F. PAPAVASILIOU, et al. (1999)
Cold Spring Harb Symp Quant Biol 64, 207-210
   Abstract »    PDF »
Analyses of Single B Cells by Polymerase Chain Reaction Reveal Rearranged VH with Germline Sequences in Spleens of Immunized Adult Rabbits: Implications for B Cell Repertoire Maintenance and Renewal.
D. Sehgal, E. Schiaffella, A. O. Anderson, and R. G. Mage (1998)
J. Immunol. 161, 5347-5356
   Abstract »    Full Text »    PDF »
Rapid elimination of mature autoreactive B cells demonstrated by Cre-induced change in B cell antigen receptor specificity in vivo.
K.-P. Lam and K. Rajewsky (1998)
PNAS 95, 13171-13175
   Abstract »    Full Text »    PDF »
Antigen Receptor Engagement Turns off the V(D)J Recombination Machinery in Human Tonsil B Cells.
E. Meffre, F. Papavasiliou, P. Cohen, O. de Bouteiller, D. Bell, H. Karasuyama, C. Schiff, J. Banchereau, Y.-J. Liu, and M. C. Nussenzweig (1998)
J. Exp. Med. 188, 765-772
   Abstract »    Full Text »    PDF »
Surrogate Light Chain Production During B Cell Differentiation: Differential Intracellular Versus Cell Surface Expression.
Y.-H. Wang, J. Nomura, O. M. Faye-Petersen, and M. D. Cooper (1998)
J. Immunol. 161, 1132-1139
   Abstract »    Full Text »    PDF »
Expression of Recombination Activating Genes in Germinal Center B Cells: Involvement of Interleukin 7 (IL-7) and the IL-7 Receptor.
M. Hikida, Y. Nakayama, Y. Yamashita, Y. Kumazawa, S.-I. Nishikawa, and H. Ohmori (1998)
J. Exp. Med. 188, 365-372
   Abstract »    Full Text »    PDF »
Restricted Expression of E2A Protein in Primary Human Tissues Correlates with Proliferation and Differentiation.
M. N. Rutherford and D. P. LeBrun (1998)
Am. J. Pathol. 153, 165-173
   Abstract »    Full Text »    PDF »
Ku70 Is Required for Late B Cell Development and Immunoglobulin Heavy Chain Class Switching.
J. P. Manis, Y. Gu, R. Lansford, E. Sonoda, R. Ferrini, L. Davidson, K. Rajewsky, and F. W. Alt (1998)
J. Exp. Med. 187, 2081-2089
   Abstract »    Full Text »    PDF »
Kinetics of Establishing the Memory B Cell Population as Revealed by CD38 Expression.
A. Ridderstad and D. M. Tarlinton (1998)
J. Immunol. 160, 4688-4695
   Abstract »    Full Text »    PDF »
Oligoclonal Development of B Cells Bearing Discrete Ig Chains in Chicken Single Germinal Centers.
H. Arakawa, K.-i. Kuma, M. Yasuda, S. Furusawa, S. Ekino, and H. Yamagishi (1998)
J. Immunol. 160, 4232-4241
   Abstract »    Full Text »    PDF »
The Normal Counterpart of IgD Myeloma Cells in Germinal Center Displays Extensively Mutated IgVH Gene, Cµ-Cdelta Switch, and lambda  Light Chain Expression.
C. Arpin, O. de Bouteiller, D. Razanajaona, I. Fugier-Vivier, F. Briere, J. Banchereau, S. Lebecque, and Y.-J. Liu (1998)
J. Exp. Med. 187, 1169-1178
   Abstract »    Full Text »    PDF »
Frequent occurrence of deletions and duplications during somatic hypermutation: Implications for oncogene translocations and heavy chain disease.
T. Goossens, U. Klein, and R. Kuppers (1998)
PNAS 95, 2463-2468
   Abstract »    Full Text »    PDF »
Rearrangement of lambda  Light Chain Genes in Mature B Cells In Vitro and In Vivo. Function of Reexpressed Recombination-activating Gene (RAG) Products.
M. Hikida and H. Ohmori (1998)
J. Exp. Med. 187, 795-799
   Abstract »    Full Text »    PDF »
V(D)J Recombinase Activity in a Subset of Germinal Center B Lymphocytes.
S. Han, S. R. Dillon, B. Zheng, M. Shimoda, M. S. Schlissel, and G. Kelsoe (1997)
Science 278, 301-305
   Abstract »    Full Text »


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