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9 B Cell Antigen Receptor Drives Generation of CD5+ B Cells Both In Vivo and In Vitro1
*
Division of Basic Sciences, Department of Pediatrics, National Jewish Medical and Research Center, Denver, CO 80206;
Department of Immunology, University of Colorado Health Sciences Center, Denver, CO 80206;
Department of Microbiology and Immunology, Shimane Medical University, Izumo, Shimane, Japan,
§
Department of Immunology, The Scripps Research Institute, La Jolla, CA 92037; and
¶
Institute for Cancer Research, Fox Chase Cancer Center, Philadelphia, PA 19111
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Abstract |
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 Top Abstract IntroductionMaterials and MethodsResultsDiscussionReferences |
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) originally isolated from a
CD5+ (B-1a) B cell generates B-1 phenotype cells in bone
marrow cultures that show a distinctive B-1 function, survival in
culture. Despite their autoreactivity, we find no evidence for receptor
editing or that the paucity of B-2 cells is the result of
tolerance-induced selection. Finally, Ca2+ mobilization
studies reveal a difference between transgenic B-1 cells in spleen and
peritoneal cavity, with cells in spleen much more responsive to
anti-B cell receptor cross-linking. We discuss these results in
terms of specificity vs lineage models for generation of distinctive B
cell subpopulations. |
Introduction |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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light chain usage (11). In
neonatal spleen, B-1 cells are relatively more abundant
(12). Besides their phenotypic distinctions, B-1 cells are
somewhat larger than B-2 cells and, when tested in vitro, differ from
B-2 cells in their responsiveness to a number of stimuli
(13, 14, 15, 16). Studies indicate that B-1 cells also contribute
disproportionately to the levels of serum IgM
(17, 18, 19). Comparison of expressed Ag receptors of B-1 and B-2 cells (BCR)3 suggests that these cell types use a partly overlapping repertoire of gene segments, but may differ in Ag specificity. Many B-1 cells appear to have germline-encoded self-reactive BCR (20, 21, 22, 23, 24, 25, 26), a property that may be specifically selected (27, 28, 29, 30, 31, 32). Although B-1 cells are subject to tolerance (25, 33, 34, 35), their threshold for tolerance induction may differ from that of B-2 cells, which are highly tolerance sensitive even at low binding affinities (36, 37, 38).
Phosphatidylcholine (PtC)-reactive Ab specificities are highly
represented in the normal B-1 cell population, where they represent
5–10% of the total. They were initially identified as
conferring reactivity to bromelain-treated mouse RBC, and such Abs
predominantly use VH11 or
VH12 genes with restricted CDR3 structure
(30, 32, 39, 40, 41, 42). Interestingly, in B-1 cells
VH11 and VH12 heavy chains
pair predominantly with V9 and
V4 light chains, respectively.
There has been much debate concerning the origins of B-1 and B-2 cells. Several lines of evidence suggest that B-1 cells are derived during a distinctive fetal/neonatal phase of B lymphopoiesis. For example, in contrast with adult bone marrow (BM) pro-B cells, fetal pro-B cells do not express TdT or precursor lymphocyte regulated myosin-like light chain, show delayed expression of class II molecules (I-A and I-E), and, when transferred to adoptive recipients, mature primarily to the B-1 phenotype (43, 44, 45, 46, 47, 48, 49, 50, 51).
On the other hand, adult BM-derived B cells with a B-2 phenotype can be induced to express a B-1 phenotype when appropriately stimulated (52, 53, 54, 55, 56). Furthermore, B-1 cells appear uniquely sensitive to mutations that alter BCR signaling, implicating such signaling in their generation or survival (3, 26, 57, 58, 59). Such studies have led to the proposal of lineage vs specificity models for B-1 cell generation (60, 61), although some of these differences may now be reconciled by postulating that fetal and adult B cell precursors represent developmental lineages that differ in their expressed Ag receptor repertoires and, hence, their specificities (53, 62, 63). This can occur through the rearrangement process, which is biased in fetal cells compared with that in adult BM precursors as a result of the absence of TdT expression (49, 60, 64, 65, 66), nonrandom V gene recombination, and altered heavy chain-selective mechanisms (63).
However, it remains to be determined whether differences in early
stages of B cell development completely account for the fetal-biased
generation of B-1 B cells or, instead, whether differences in BCR
selection/tolerance also play a role. Therefore, in this report we test
whether B cells with a transgenic B-1 BCR experience tolerance in BM
differently from transgenic B-2 BCR B cells. Our work was prompted by
the general observation that most Ig transgenic mice are deficient in
B-1 cells, and that B-1 cells present in such mice usually express
endogenous Ig chains (67, 68, 69). In contrast, in two Ig-Tg
mice expressing B-1 B cell-derived receptors reactive to PtC, B-1 B
cells are abundant and express the transgenic specificity, whereas B-2
cells are less abundant (63, 69). Specifically, we have
followed the fate of B cells generated in the BM of
VH11/V9 Tg mice,
comparing it with that in 3-83 µ
(anti-H-2Kk,b) Ig-transgenic mice (70, 71). In this
VH11V9 transgenic model,
we find no evidence for immune tolerance or receptor editing, so that
self-reactive B cells are produced and populate a peripheral B-1 pool.
These cells show persistence in culture, a function shared with
normally generated B-1 B cells. Furthermore, these cells are not
anergic, producing large amounts of serum Ig and showing a calcium flux
in response to receptor cross-linking.
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Materials and Methods |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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3-83 µ transgenic mice were maintained on the
autoantigen-free, nondeleting (H-2Kd) background
of B10.D2nSn/J (The Jackson Laboratory, Bar Harbor, ME) or on the
autoantigen-containing, deleting background
(H-2Kk) of B10.BR (provided by Pippa Marrack,
National Jewish Medical and Research Center, Denver, CO).
VH11V9 transgenic mice,
originally on the CB17 background, were backcrossed at least six times
on the B10.D2nSn/J background. Mice were housed and bred at the
National Jewish Medical and Research Center Biological Research
facility and were used at 8–12 wk of age.
Cell culture
BM cells were isolated from aseptically removed femurs and grown
in vitro as previously described (72, 73). Briefly, BM
cells were depleted of RBCs and cultured at a concentration of 2
x 106 cells/ml in IMDM (Life Technologies,
Gaithersburg, MD) supplemented with 10% FCS (Gemini Bio-Products,
Calabasas, CA), 5 x 10-5 M 2-ME, 1%
sodium pyruvate, 100 U/ml penicillin, 100 µg/ml streptomycin, and 2
mM L-glutamine at 37°C with 7%
CO2. Primary BM cultures were supplemented with
50–100 U/ml of rIL-7, which preferentially expanded IL-7-responsive B
cell precursors, and after 5–6 days the cultures contained >98%
B220+ cells (72). Where noted, BM
cells were incubated at 4°C for 1 h with rat
anti-mIgM-biotin (b-7-6) precoupled to M-280 streptavidin magnetic
beads (Dynal, Lake Success, NY), and the IgM+
cells were removed magnetically. Purity was assessed by
immunofluorescence, and in all cases the IgM-
population was >99% pure. Twenty-four-hour and 13-day cultures
(secondary cultures) of BM were performed without the addition of IL-7.
Splenic cells were depleted of erythrocytes using buffered ammonium
chloride. B cells were purified by T cell depletion with anti-Thy-1
mAb and rabbit complement (Cedarlane, Westbury, NY), followed by
density separation (
= 1.066) through Percoll
(Amersham-Pharmacia, Piscataway, NJ). Purified B cells were cultured as
described above. Peritoneal cells were removed by aseptic injection of
10 ml of IMDM (Life Technologies) into the peritoneal cavity followed
by withdrawal of the peritoneal exudate. Where noted, 10 µg/ml LPS
(Life Technologies), 10 µg/ml rat anti-mIg (187.1) mAb (a gift
from Dr. J. Cambier, National Jewish Medical and Research Center), 10
µg/ml rat anti-azophenylarsenate (5 Ci) mAb (a gift from Dr. L.
Wysocki, National Jewish Medical and Research Center), or 10 µg/ml
PtC-containing liposomes (24) were added to the
cultures.
Flow cytometry
mAbs used for mouse cell staining included anti-B220,
RA3–6B2; anti-CD23, B3B4; anti-CD43, S7; anti-CD5, 53-7;
anti-IgMa, RS3.1; anti-Id for 3-83, 54.1
(70); and anti-Id for VH11, RidA
(33). Fluorescent reagents were either purchased
(PharMingen, San Diego, CA) or labeled as previously described
(4). Polyclonal FITC-anti-Ig light chain was
purchased from Southern Biotechnology (Birmingham, AL). Biotinylated
Abs were visualized with streptavidin-Tri-Color (Caltag, South San
Francisco, CA) for three-color analysis on a FACSCalibur (Becton
Dickinson, San Jose, CA) or with Texas Red-avidin for four-color
analysis on a dual laser dye laser FACStarPlus
(Becton Dickinson). Forward and side scatter gates were adjusted to
include only nucleated viable cells.
Cell survival
Before all analyses of cell survival, cells were counted on a hemocytometer using trypan blue exclusion, and the frequency of B220+ cells was determined by immunofluorescence. Cultures were initiated at 2 x 106 cells/well in six-well culture plates. After incubation, wells were harvested and counted, and the frequency of B220+ cells was determined by immunofluorescence. Cell survival is reported for B220+ cells as a percentage of their initial numbers.
Carboxyfluorescein diacetate succimidylester (CFSE) labeling
Cells treated with CFSE (Molecular Probes, Eugene, OR) were labeled as previously described (74). Briefly, a stock solution of 5 mM CFSE in DMSO was diluted 1/10 in serum-free IMDM prewarmed to 37°C. B cells were diluted to 50 x 106 cells/ml and incubated with 10 µl of diluted CFSE/ml at 37°C for 10 min. Labeled cells were washed twice with IMDM and placed in culture as described above.
BM chimeras
BM chimeras were prepared as previously described (75), but with the following alterations. Ten- to 12-wk-old donor femurs were removed aseptically, and B cells were isolated as described above. The purified cells were then incubated with M280-streptavidin magnetic beads (Dynal) conjugated to anti-B220-biotin (RA3-6B2) and anti-CD19-biotin (1D3, PharMingen) mAbs for 45 min at 4°C. Bound cells were removed magnetically, and the remaining mixture was assessed for purity (B220-, CD19-) by immunofluorescence. A purity level of >99% B220-, CD19- was usually attained following just one round of isolation, but in some cases the process was repeated to achieve this level of purity. The viable cell concentration following the removal of B220+, CD19+ cells was adjusted to 5 x 107 cells/ml in sterile HBSS (Life Technologies), and 0.1 ml was injected i.v. per irradiated recipient. Recipients were age-matched B10.D2nSn/J mice that received 960 rad whole-body irradiation at least 1 h before BM transfer. Recipient mice were analyzed 4–6 wk after transfer.
Detection of RAG-2 mRNA
mRNA was isolated from 3–5 x 106 freshly isolated or cultured BM cells using the Oligotex Direct mRNA isolation kit according to the manufacturer’s instructions (Qiagen, Valencia, CA). First-strand cDNA synthesis was conducted by random priming of 1 µg of isolated mRNA using a kit (Ready-to-Go, Amersham-Pharmacia) in a volume of 20 µl. PCR amplification of 1 µl of the resulting cDNA was conducted as previously described (72) using primers specific for CD19 and RAG-2. PCR reactions were electrophoresed in 1% agarose, transferred to a Zeta-Probe membrane (Bio-Rad, Richmond, CA), and hybridized overnight with 32P-labeled CD19- and RAG-2-specific probes. Autoradiography was performed as described previously (72).
ELISA
Ig concentrations were measured as previously described (76). Briefly, Immulon 2HB polystyrene plastic microtiter plates (Dynex Technologies, Chantilly, VA) were coated with rat anti-mIgM (b-7-6), washed, and blocked. Sera (diluted in PBS supplemented with 1% BSA) were incubated for 3 h at 25°C, and bound Ig was detected with biotinylated rat anti-mIgM (Ak2) or rat a-mIgMa (RS3.1) and streptavidin-peroxidase (Sigma, St. Louis, MO). Standard curves for the assay were generated using COS line D1 supernatant. Absorbance was measured on a Bio-Rad model 2225 ELISA plate reader (Bio-Rad) and analyzed using the Microplate Manager program.
Analysis of calcium mobilization
Splenic and peritoneal B cells were isolated and purified as previously described and subsequently loaded with indo-1/AM (Molecular Probes) for 45 min at 37°C. Cells were washed twice, resuspended at a concentration of 106 cells/ml in IMDM and 5% FCS, and stimulated with F(ab')2 rabbit anti-mouse Ig (Zymed, San Francisco, CA). Mean fluorescence was evaluated using an Ortho model 50H flow cytometer (Ortho, Westwood, MA) and an appended data acquisition system and MultiTime software (Phoenix Flow Systems, San Diego, CA).
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Results |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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9 transgenic B cells have a B-1,
and 3-83 µ B cells have a B-2 phenotype
The cell surface marker phenotype of B cells isolated from
VH11V9, 3-83 µ
Ig-Tg, and nontransgenic mice, as analyzed by flow cytometry, is shown
in Fig. 1
. The majority of
B220+ B cells in 3-83 and wild-type spleens have
a conventional B-2 phenotype: B220high,
CD23+, CD43-,
CD5-. In contrast, splenic B cells of the
VH11V9 mouse have a
distinctive B-1 phenotype: B220low,
CD23-, CD43+,
CD5+. The peritoneal cavity B cell compartment,
which in the nontransgenic B10.D2 is primarily composed of B-1 cells,
contains 
25-fold fewer B cells in the 3-83 mouse, the majority of
which are 3-83 id- (Table I). In contrast, the
VH11V9 Tg mouse
peritoneal cavity contains 7-fold more B cells than the nontransgenic
littermate, and nearly all are B-1 cells that express the
VH11V9 receptor.
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9 Ig-transgenic B cells exhibit
enhanced in vitro survival
To further test the phenotype of the transgenic B cells, we
measured the ability of isolated
VH11V9, 3-83, or
nontransgenic splenic B cells to survive in vitro, because unlike B-2
cells, B-1 cells survive in vitro for extended periods
(77). After 7 days in culture in standard medium
containing serum but no added mitogens, >50% of the
VH11V9+
B cells were viable (Fig. 2A),
whereas nearly all the 3-83+ or nontransgenic
splenic B cells died. The enhanced in vitro survival of the
VH11V9+
splenic B cells was not associated with proliferation, as CFSE-labeled
VH11V9+
cells maintained their label during 5 days of culture (Fig. 2B). When the mitogen LPS was included in the culture
medium,
VH11V9+
splenic B cells were capable of proliferating, as demonstrated by the
loss of CFSE fluorescence. These data demonstrate that
VH11V9 Tg splenic B
cells acquire not only the characteristic cell surface phenotype of B-1
cells, but also a characteristic functional property, cell culture
survival, while 3-83+ splenic B cells display a
B-2-like phenotype and functional response.
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9 Ig-transgenic precursors in adult
BM generate B-1 B cells after transfer
To test whether the B-1 B cells in the
VH11V9 Tg mouse could be
generated from BM, we transferred
B220-/CD19- BM cells from
10- to 12-wk-old VH11V9
or 3-83 mice into lethally irradiated B10.D2 hosts and then analyzed
the phenotype of reconstituted peripheral B cells 4–6 wk later.
Donor-derived cells were identifiable by transgene Id (Fig. 3A). Recipients of
VH11V9 and 3-83 cells
contained similar numbers of donor-derived splenic B cells, but their
cell surface phenotypes differed, with
VH11V9 BM-reconstituted
mice exhibiting a B-1 phenotype and 3-83 BM recipients exhibiting a B-2
phenotype (data not shown). In mixed
VH11V9/3-83 BM
transfers, B cells expressing either the
VH11V9 BCR or the 3-83
BCR were produced (Fig. 3A) and maintained their respective
B-1-like and B-2-like phenotypes (Fig. 3B), indicating that
VH11V9+
B-1 cells do not suppress the development of 3-83 B-2 cells and
vice versa. Furthermore, splenic B cells from
VH11V9 BM-reconstituted
mice had an extended life span in vitro, whereas B cells of
3-83-reconstituted mice did not (Fig. 3C). Thus, in BM
chimeras, splenic B cell surface and survival phenotypes recapitulated
those of transgenic donor mice.
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9+ BM generates B cells
that have a B-1 phenotype and enhanced survival
To probe the differentiation of 3-83 and
VH11/V9 BM B cells in
more detail, BM precursors were grown in IL-7-containing medium, which
allows the expansion of Ig transgene-positive cells and, when Ag is
present, the investigation of tolerance mechanisms. After 5 days of
culture in the presence of IL-7 (primary cultures) B cell numbers
increased substantially (Fig. 4A), while other cell types
were lost. As in the radiation chimeras, essentially all
VH11V9+
B cells generated in IL-7 cultures were CD5+,
CD23-, and CD43+, whereas
B cells generated from 3-83 BM lacked expression of CD5 and CD43 (Fig. 4B). Unlike in vivo differentiation, the 3-83 B cells
generated in vitro did not express CD23. This difference is probably
due to the inability of IL-7-driven BM cultures to progress beyond the
immature B cell stage, which is normally CD23-.
When these cells were replated in the absence of IL-7 the
VH11V9+
B cells displayed enhanced in vitro survival, similar to that seen with
in situ generated cells, whereas the 3-83+ and
nontransgenic B cells died rapidly, after an initial 2-day lag (Fig. 4C). CFSE analysis again demonstrated that the enhanced
recovery of
VH11V9+
cells was due to prolonged survival, rather than proliferation (data
not shown). This observation strongly suggests that the B-1 phenotype
can be rapidly acquired by newly developing BM B cells in vitro,
provided that they bear a B-1-derived Ag receptor.
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9 Ig-transgenic B cells do
not spontaneously edit in the BM
To further assess the susceptibility of
VH11V9 transgenic B
cells to tolerance in the BM, we investigated the level of receptor
editing in these cells. First, we measured BM RAG-2 mRNA levels using a
semiquantitative RT-PCR assay, normalizing RNA levels using the
pan-specific B cell marker gene CD19. As shown in Fig. 5A, 3-83 and
VH11V9 Ig-transgenic BM
had low levels of RAG-2 mRNA, whereas a high level was detected in
Ag-containing 3-83/H-2Kk mouse BM and control
B10.D2 BM. As a second indicator of receptor editing, we measured the
frequencies of B cells bearing Ig light chains. In agreement with
the RAG-2 mRNA analysis, 3-83 and
VH11V9 Ig-transgenic BM
B cells had a low frequency of Ig + cells,
while 3-83/H-2Kk mouse BM cells had significantly
higher frequencies of Ig + cells (data not
shown). Finally, to determine whether
VH11V9 BM B cells were,
in fact, capable of receptor editing, we treated freshly isolated BM
cells with anti- mAb and again measured RAG-2 mRNA levels using
RT-PCR and Ig light chain expression. In BM samples from
VH11V9, 3-83, and
nontransgenic control mice, cross-linking the BCR with anti- mAb
resulted in a detectable induction of RAG-2 (Fig. 5B) and
elevated Ig light chain expression (data not shown), whereas
treatment with control rat mAb did not. Similar enhanced RAG-2 and Ig
light chain expression were seen following coculture with
PtC-containing liposomes (data not shown).
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9+ splenic
B cells, but not peritoneal B cells
Given the self-reactivity of the
VH11V9+
cells (27, 30), we wondered whether the peripheral cells
with this BCR were functional or tolerized. Two lines of evidence
suggested that they were not anergic. First, the
VH11V9 mice secreted
abundant amounts of this Ab into the serum, indicating that their B
cells had effector function (Fig. 6A). Second, unlike anergic B
cells, spleen cells derived from
VH11V9 mice had robust
Ca2+ mobilization responses upon receptor
ligation with F(ab')2 rabbit anti-mouse IgM
(Fig. 6B). In contrast, peritoneal B cells from these same
mice did not respond, suggesting an environmental influence on their
function. These data indicate that the
VH11V9 BCR on splenic B
cells is functional and that these B cells are not anergic despite the
presence of PtC in the environment.
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Discussion |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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Although the simplest lineage model of B-1 vs B-2 development
(61) would not have predicted the results reported here,
the clear-cut difference in the abilities of fetal and BM pro-B cells
to reconstitute B-1 and B-2 development remain (45, 47).
Similarly, these data, while consistent with the ability of the BCR to
induce a B-1 phenotype (29), do not obscure the fact that
fetal and adult precursors show various distinctions in gene expression
(49) and also differ markedly in their ability to give
rise to these B cell types. In this study we have only compared the
development of BM-derived precursors that carried either B-1-derived or
B-2-derived transgenic BCRs. One way to put the present results into
context is to consider the role of cell lineage in regulating the
preselected BCR repertoire. Several lines of evidence suggest that the
VH11 H chain used in this study is normally
counterselected in BM compared with fetal precursors due to its poor
ability to associate with the surrogate L chain components 5 and
VpreB (63). The particular VH11 and
V9 transgenes used in our study force B-1
receptor expression in cells that normally disfavor its generation,
propagation, and survival, allowing a test of the effect of such a
receptor on further B cell development, but obscuring the effect of
this receptor on earlier developmental stages. This idea is
consistent with the view that B-1 receptors are unusual,
germline-encoded specificities that are beneficial (81)
and therefore are selected over evolution for particular properties
that promote their generation in the fetus, but disfavor their
development in the adult (63).
Do our results demonstrate that any
VH11V9+
B cell generated in the BM of normal (nontransgenic) mice will enter
the peripheral B-1 B cell pool? Possibly, but one must also consider
that differences in the level of expression of Ag receptor can have a
striking effect on B cell development (82). The particular
VH11-µ transgenic mouse line used in our study,
BR1, was selected for its capacity to suppress RAG expression and
therefore block endogenous heavy chain rearrangements. This line has a
distinctively higher level of µ protein expression compared with
normal B cells, probably due to the high transgene copy number
(63). A different VH11-µ
transgenic line, BR5, shows a lower level of heavy chain expression
that may be more physiologic. Therefore, it will be important in future
work to perform similar experiments using the BR5 line to generalize
our results to normal VH11 development and test
whether BCR levels alter selection. Furthermore, considering the
distinctions mentioned above that distinguish fetal liver from BM B
lymphopoiesis, it will also be important to compare cells isolated from
BM with those obtained from fetal liver.
The inability of
VH11V9+
B cells isolated from the peritoneal cavity to mobilize
Ca2+ following BCR ligation suggests an anergic
phenotype for these cells. However,
VH11V9+
splenic B cells are normally responsive to the same stimulus, and the
presence of significant levels of
VH11V9 serum Ig suggests
a nonanergic phenotype. We are currently pursuing the idea that
differences in the microenvironment between spleen and peritoneal
cavity, either Ag availability or cytokine milieu, result in this
differential responsiveness.
A key biological response that appears to be controlled by the quality
of the BCR signal is B cell survival. Studies of immunological
tolerance over the years have clearly shown that in many situations,
reactivity to self-tissues radically alters the survival of B cells in
the peripheral immune system, leading to outright deletion or shortened
lifespan compared with nonautoreactive B cells (76, 83, 84, 85). On the other hand, an insufficiency of BCR-mediated
signaling, such as occurs upon the genetic elimination of key BCR
coreceptors or molecules in the signaling pathway (Btk, CD45, CD19,
Syk), also blocks developmental progression and the acquisition of a
long B cell lifespan (3, 58, 59, 86, 87, 88, 89). Furthermore, in
B-2 cells, continued BCR expression is required for survival even after
recruitment into the long-lived pool (90). The in vivo
situation with B-1 cells is more complex, as these cells appear to
require Ag for their selection or persistence (or both) and can undergo
chronic receptor-driven expansion (62, 69, 91). In the
present study we document the extended in vitro survival of B-1
phenotype cells, relative to B-2 cells. Taking advantage of this
knowledge to probe B cell differentiation, we find that a long-lived
phenotype is rapidly conferred to newly formed, BM B cells expressing a
B-1-derived, but not a B-2-derived, receptor. This differential
survival in vitro correlates with the B-1 phenotype of the splenic B
cells of VH11V9 Ig
transgenic mouse and the B-2 phenotype of 3-83 Tg mouse B cells.
Although it is still unclear how B-1 and B-2 cells of different
specificities survive in vitro, it is tempting to speculate that this
might point to a general difference in the survival of these two B cell
types. This experimental system should be very helpful in the future in
determining more precisely how the BCR controls cell survival.
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Acknowledgments |
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Footnotes |
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2 Address correspondence and reprint requests to Dr. Richard R. Hardy, Institute for Cancer Research, Fox Chase Cancer Center, 7701 Burholme Avenue, Philadelphia, PA 19111.
3 Abbreviations used in this paper: BCR, B cell Ag receptor; PtC, phosphatidylcholine; BM, bone marrow; CFSE, carboxyfluorescein diacetate succimidylester; RAG-2, recombination-activating gene-2.
Received for publication December 3, 1999. Accepted for publication February 24, 2000.
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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2 in B-1 lymphocytes. Int. Immunol. 6:1011.-deficient animals: a potential mechanism of B cell tolerance. J. Exp. Med. 180:1805.This article has been cited by other articles:
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G. F. Widhopf II, D. C. Brinson, T. J. Kipps, and H. Tighe Transgenic Expression of a Human Polyreactive Ig Expressed in Chronic Lymphocytic Leukemia Generates Memory-Type B Cells That Respond to Nonspecific Immune Activation J. Immunol., February 15, 2004; 172(4): 2092 - 2099. [Abstract] [Full Text] [PDF]
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K. Balabanian, J. Couderc, L. Bouchet-Delbos, A. Amara, D. Berrebi, A. Foussat, F. Baleux, A. Portier, I. Durand-Gasselin, R. L. Coffman, et al. Role of the Chemokine Stromal Cell-Derived Factor 1 in Autoantibody Production and Nephritis in Murine Lupus J. Immunol., March 15, 2003; 170(6): 3392 - 3400. [Abstract] [Full Text] [PDF]
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 K. Hayakawa, M. Asano, S. A. Shinton, M. Gui, L.-J. Wen, J. Dashoff, and R. R. Hardy Positive Selection of Anti-Thy-1 Autoreactive B-1 Cells and Natural Serum Autoantibody Production Independent from Bone Marrow B Cell Development J. Exp. Med., January 6, 2003; 197(1): 87 - 99. [Abstract] [Full Text] [PDF]
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 S.-C. Wong, W.-K. Chew, J. E.-L. Tan, A. J. Melendez, F. Francis, and K.-P. Lam Peritoneal CD5+ B-1 Cells Have Signaling Properties Similar to Tolerant B Cells J. Biol. Chem., August 16, 2002; 277(34): 30707 - 30715. [Abstract] [Full Text] [PDF]
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M. J. Chumley, J. M. Dal Porto, and J. C. Cambier The Unique Antigen Receptor Signaling Phenotype of B-1 Cells Is Influenced by Locale but Induced by Antigen J. Immunol., August 15, 2002; 169(4): 1735 - 1743. [Abstract] [Full Text] [PDF]
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S. A. Johnson, S. J. Rozzo, and J. C. Cambier Aging-Dependent Exclusion of Antigen-Inexperienced Cells from the Peripheral B Cell Repertoire J. Immunol., May 15, 2002; 168(10): 5014 - 5023. [Abstract] [Full Text] [PDF]
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T. L. Rothstein Two B-1 or Not To Be One J. Immunol., May 1, 2002; 168(9): 4257 - 4261. [Abstract] [Full Text] [PDF]
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M. I. D. Rossi, K. L. Medina, K. Garrett, G. Kolar, P. C. Comp, L. D. Shultz, J. D. Capra, P. Wilson, A. Schipul, and P. W. Kincade Relatively Normal Human Lymphopoiesis but Rapid Turnover of Newly Formed B Cells in Transplanted Nonobese Diabetic/SCID Mice J. Immunol., September 15, 2001; 167(6): 3033 - 3042. [Abstract] [Full Text] [PDF]
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M. Kraus, L. I. Pao, A. Reichlin, Y. Hu, B. Canono, J. C. Cambier, M. C. Nussenzweig, and K. Rajewsky Interference with Immunoglobulin (Ig){alpha} Immunoreceptor Tyrosine-Based Activation Motif (Itam) Phosphorylation Modulates or Blocks B Cell Development, Depending on the Availability of an Ig{beta} Cytoplasmic Tail J. Exp. Med., August 20, 2001; 194(4): 455 - 470. [Abstract] [Full Text] [PDF]
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S.-H. Park, A. Weiss, K. Benlagha, T. Kyin, L. Teyton, and A. Bendelac The Mouse Cd1d-Restricted Repertoire Is Dominated by a Few Autoreactive T Cell Receptor Families J. Exp. Med., April 16, 2001; 193(8): 893 - 904. [Abstract] [Full Text] [PDF]
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H. H. Wortis and R. Berland Cutting Edge Commentary: Origins of B-1 Cells J. Immunol., February 15, 2001; 166(4): 2163 - 2166. [Abstract] [Full Text] [PDF]
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U. Kim, C. S. Gunther, and R. G. Roeder Genetic Analyses of NFKB1 and OCA-B Function: Defects in B Cells, Serum IgM Level, and Antibody Responses in Nfkb1-/-Oca-b-/- Mice J. Immunol., December 15, 2000; 165(12): 6825 - 6832. [Abstract] [Full Text] [PDF]
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 P. Sanchez, A.-M. Crain-Denoyelle, P. Daras, M.-C. Gendron, and C. Kanellopoulos-Langevin The level of expression of {micro} heavy chain modifies the composition of peripheral B cell subpopulations Int. Immunol., October 1, 2000; 12(10): 1459 - 1466. [Abstract] [Full Text] [PDF]
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B10. D2, 3-83 
