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RP105-lacking B cells from lupus patients are responsible for the production of immunoglobulins and autoantibodies.

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ARTHRITIS & RHEUMATISM
Vol. 46, No. 12, December 2002, pp 3259–3265
DOI 10.1002/art.10672
© 2002, American College of Rheumatology
RP105-Lacking B Cells From Lupus Patients Are
Responsible for the Production of
Immunoglobulins and Autoantibodies
Yuji Kikuchi,1 Syuichi Koarada,2 Yoshifumi Tada,2 Osamu Ushiyama,2 Fumitaka Morito,2
Noriaki Suzuki,2 Akihide Ohta,2 Kensuke Miyake,2 Masao Kimoto,2
Takahiko Horiuchi,3 and Kohei Nagasawa2
Objective. We previously reported that B cells
lacking the RP105 molecule, which proved to be highly
activated B cells, are increased in the peripheral blood
of patients with systemic lupus erythematosus (SLE). In
the present study, we attempted to determine whether
RP105-negative B cells obtained from SLE patients
would be capable of producing autoantibodies as well as
immunoglobulins.
Methods. RP105-positive and RP105-negative B
cells, sorted by cell sorter, were cultured for 5 days
without stimulation, or were stimulated with Staphylococcus aureus Cowan 1 strain (SAC) or recombinant
interleukin-6 (IL-6). For the assay of autoantibodies,
RP105-positive and RP105-negative B cells were cultured separately for 10 days with anti-CD3 antibody–
stimulated T cells. The production of immunoglobulins
and autoantibodies was determined by enzyme-linked
immunosorbent assay.
Results. We demonstrated that RP105-negative B
cells, but not RP105-positive B cells, obtained from SLE
patients could spontaneously produce IgG and IgM in
vitro until day 5. SAC and IL-6 enhanced production of
IgG and IgM by RP105-negative B cells but failed to
induce such production by RP105-positive B cells. The
latter cells, however, when cocultured with activated T
cells in the presence of IL-10, produced IgG, although
the amount was very small compared with that produced by RP105-negative B cells. Most important, under
these conditions, anti–double-stranded DNA antibodies
were produced only by the RP105-negative B cells
obtained from SLE patients.
Conclusion. These data indicate that RP105negative B cells, constituting a subset of B cells in SLE
patients, are highly activated and may be responsible
for the production of autoantibodies as well as polyclonal immunoglobulins.
Systemic lupus erythematosus (SLE) is an autoimmune disease characterized by dysfunction of T cells
and polyclonal B cell activation, resulting in production
of pathogenic IgG autoantibodies such as anti–doublestranded DNA (anti-dsDNA) antibody. High-affinity
anti-dsDNA IgG in SLE bears characteristics of an
antigen-driven, T cell–dependent immune response.
Such autoantibodies may play a critical role in the
progression of lupus nephritis. In fact, exacerbations of
the disease are preceded by increasing levels of antidsDNA (1–4). It is known that peripheral B cells obtained from patients with SLE contain populations that
spontaneously produce immunoglobulins, including autoantibodies, and also can mature into antibodysecreting cells when cultured in vitro in the absence of
obvious activators of B cell differentiation (5–7).
In vivo, antigen-specific activation and differentiation of B cells occur in germinal centers in the
lymphoid system, where naive B cells undergo activation, proliferation, somatic hypermutation of rearranged
V region genes, immunoglobulin isotype switching, and
Supported in part by grant-in-aid 13670459 from the Ministry
of Education, Culture, Sports, Science and Technology of Japan.
1
Yuji Kikuchi, MD: Saga Medical School, Saga, Japan, and
Kyushu University Graduate School of Medical Sciences, Fukuoka,
Japan; 2Syuichi Koarada, MD, Yoshifumi Tada, MD, Osamu Ushiyama,
MD, Fumitaka Morito, MD, Noriaki Suzuki, MD, Akihide Ohta, Kensuke Miyake, MD, Masao Kimoto, MD, Kohei Nagasawa, MD: Saga
Medical School, Saga, Japan; 3Takahiko Horiuchi, MD: Kyushu University Graduate School of Medical Sciences, Fukuoka, Japan
Address correspondence and reprint requests to Yuji Kikuchi,
MD, Department of Medicine and Biosystemic Science, Kyushu
University, Graduate School of Medical Sciences, Fukuoka 812-8582,
Japan. E-mail: Kikuchin@intmed1.med.kyushu-u.ac.jp.
Submitted for publication March 5, 2002; accepted in revised
form August 29, 2002.
3259
3260
KIKUCHI ET AL
subsequent positive and negative selection by antigen.
Moreover, activated B cells mature into antibodyproducing plasma cells, or, alternatively, become memory B cells within germinal centers (8–11).
RP105, a novel molecule on B cells identified in
1994 (12), transmits an activation signal that leads to B
cell proliferation and protection from apoptosis in mice.
The extracellular domain of RP105 is structurally similar
to that of Toll-like receptors, suggesting that RP105
senses pathogen invasion and activates B cells (13–15).
Mice devoid of RP105 were recently generated, and it
has been shown that RP105 regulates lipopolysaccharide
signaling in B cells (16). In contrast to these findings in
mice, little is known about RP105 in humans: the human
homologue of RP105 has been identified and its monoclonal antibody established (17,18). Histologic studies
showed that RP105 was expressed mainly on mature B
cells in mantle zones, and that germinal center cells were
negative for RP105 (19).
It is known that virtually all mature B cells have
RP105 molecules on their surface, in humans as well as
in mice (19). We previously demonstrated that the
number of RP105-negative B cells is significantly increased in the peripheral blood of patients with SLE,
Sjögren’s syndrome, and dermatomyositis, the pathophysiology of which is characterized by highly activated
B cells (20–22). Particularly in SLE, the increase in
RP105-negative B cells was well correlated with disease
activity (20). We defined the phenotype of RP105negative B cells as CD95-positive, CD86-positive, and
CD38-bright, which is consistent with that of activated B
cells or germinal center B cells. Moreover, RP105negative B cells had intracellular IgG, suggesting that
they could produce IgG (20).
It was of great interest, therefore, to determine
whether RP105-negative B cells obtained from patients
with SLE would be responsible for the production of
autoantibodies. In this study, we demonstrated that
RP105-negative B cells, but not RP105-positive B cells,
produced immunoglobulins without stimulation and
even produced anti-dsDNA antibody, with activated T
cell help.
PATIENTS AND METHODS
Patients. Seven patients who had active SLE and were
attending our hospital were enrolled in this study. All patients
were female and met the American College of Rheumatology
criteria for SLE (23). The activity of SLE was assessed by the
SLE Disease Activity Index (SLEDAI) (24), and we defined
the active disease state as SLEDAI scores of ⱖ14. All patients
in this study had anti-dsDNA antibody in their sera. In patients
1, 2, 4, 5, and 7, SLE had not been diagnosed previously, and
the diagnosis was first made at the time of this analysis.
Although patient 3 had been on a maintenance dosage of
prednisolone (15 mg/day), progressive proteinuria occurred.
Patient 6 had been given a maintenance dosage of predonisolone (5 mg/day), but slight fever and decreased serum
level of complement emerged. Patients ranged in age from 17
to 65 years (mean 36 years).
Cell preparation. Samples of peripheral venous blood
obtained from patients with SLE were collected into tubes
containing heparin. Peripheral blood mononuclear cells
(PBMCs) were separated by centrifugation over FicollHypaque (Pharmacia Biotech, Uppsala, Sweden) gradient.
Immunofluorescence analysis of B cells. PBMCs were
stained with fluorescein isothiocyanate–conjugated antihuman RP105 monoclonal antibody (mAb) (20) and
phycoerythrin-conjugated anti-human CD19 mAb (BD
PharMingen, San Diego, CA), and analyzed using FACScan.
Statistical analyses were performed using CellQuest software
(Becton Dickinson, Mountain View, CA). Cells were then
sorted into 2 fractions, RP105-negative CD19-positive B cells,
and RP105-positive CD19-positive B cells, using the Epics
Elite cell sorter (Coulter, Hialeah, FL).
Preparation of T cells. PBMCs obtained from a
healthy donor were isolated by centrifugation over FicollHypaque (Pharmacia Biotech) gradient. CD19-negative cells
were isolated by negative selection using immunomagnetic
beads (Dynal, New Hyde Park, NY). CD19-negative cells were
allowed to adhere to plastic dishes. Nonadherent cells were
used as T cells. T cells were stimulated with 10 ␮g/ml of
phytohemagglutinin (Difco, Detroit, MI) and 100 units/ml of
recombinant interleukin-2 (IL-2) (Takeda Chemical Industries, Osaka, Japan) every 4 days. These activated T cells were
irradiated at 3,000 rad, and 2 ⫻ 105 cells were cultured in
anti-human CD3 antibody–coated 96-well plates.
Production of total IgG and IgM. RP105-positive B
cells and RP105-negative B cells were separately suspended in
RPMI 1640 containing 10% fetal calf serum and incubated in
a 5% CO2 incubator at 37°C in 96-well plates (Nalgene Nunc,
Milwaukee, WI). Cells (2 ⫻ 104) were cultured for 5 days
without stimulation, or were stimulated with 0.001% Staphylococcus aureus Cowan 1 strain (SAC), or 1 ng/ml recombinant
IL-6 (rIL-6) (Genzyme, Cambridge, MA) with or without
50 ␮g/ml anti–IL-6 antibody (Genzyme). The cultured supernatants were harvested on day 3 and day 5 and added to goat
anti-human immunoglobulin–coated (Biosource, Fleurus, Belgium) 96-well Nunc Maxisorb immunoplates (Nalgene Nunc)
with control human IgG and IgM (Chemicon, Hofheim, Germany) overnight at 4°C. After discarding the supernatants and
washing with 0.05% Tween in phosphate buffered saline
(PBS), the bound human immunoglobulin was detected with
peroxidase-labeled goat anti-human IgG ␥ chain and IgM ␮
chain (BioSource) at a dilution of 1:2,000. The plates were
washed again and developed with o-phenylendiamine. The
reaction was stopped with 3N HCl solution, and the optical
density at 490 nm was read with an ImmunoMini NJ-2300
microplate reader (System Instruments, Tokyo, Japan).
Production of IL-6. The supernatants cultured with no
stimulation were measured using an ultrasensitive IL-6
enzyme-linked immunosorbent assay (ELISA) kit (BioSource).
SIGNIFICANT ROLE OF RP105-NEGATIVE B CELLS IN SLE
Table 1. Demographic data and frequency of RP105-negative B cells
in the peripheral blood of female patients with systemic lupus
erythematosus
Patient
1
2
3
4
5
6
7
Age, years
Treatment at study
entry
% RP105-negative
B cells
17
32
26
36
39
40
65
–
–
Prednisolone, 15 mg/day
–
–
Prednisolone, 5 mg/day
–
20.8
19.9
20.0
19.0
31.0
8.8
16.7
Production of anti-DNA IgG. Monoclonal anti-human
CD3 antibody (UCHL1; Genzyme/Techne, Minneapolis, MN)
was diluted to a concentration of 1 ␮g/ml in PBS. A total of 100
␮l was added to each culture well, and the plates were
incubated for at least 2 hours at room temperature, then
washed with PBS to remove any anti-CD3 antibody that was
not bound to the plates. RP105-positive and RP105-negative B
cells were separately cultured for 10 days at a density of 5–10 ⫻
104 cells/well with 2 ⫻ 105 T cells stimulated with immobilized
anti-CD3 antibody and 10 ng/ml of recombinant IL-10
(Genzyme/Techne), and the supernatants were harvested.
Single-stranded DNA (ssDNA) was obtained by boiling calf
thymus DNA (Worthington, Freehold, NJ) for 5 minutes,
followed by rapid cooling on ice. Double-stranded DNA was
obtained by treating calf thymus DNA with S1 nuclease. The
plates were coated with 10 ␮g/ml of calf thymus DNA. The
plates were washed and blocked with PBS containing 1%
bovine serum albumin. The samples of culture supernatants
were incubated in the plates overnight at 4°C. The bound
human immunoglobulin was detected as described above. In
the serum obtained from 1 patient with SLE, the titer of
anti-dsDNA antibody was 500 IU/ml, and the titer of antissDNA antibody was 500 AU/ml; this serum was used as a
control.
3261
produced neither IgG nor IgM (detection threshold for
IgG 150 ng/well, for IgM 7.5 ng/well). IgG production by
RP105-negative B cells obtained from patients 1–3 was
not increased on day 5 (13.5 ⫾ 6.7 ␮g/ml) compared with
day 3 (13.4 ⫾ 6.2 ␮g/ml). In contrast, IgG production by
RP105-negative B cells obtained from patient 4 increased on day 5 (13.0 ␮g/ml) compared with day 3 (7.50
␮g/ml) (Figure 1). IgM production by RP105-negative B
cells from patients 1, 3, and 4 was not significantly
increased on day 5 (245.0 ⫾ 74.6 ng/ml) compared with
day 3 (213.0 ⫾ 61.8 ng/ml) (Figure 1).
The effect of SAC and rIL-6 on in vitro immunoglobulin synthesis by RP105-positive and RP105negative B cells. Addition of 0.001% SAC enhanced
total IgG production by RP105-negative B cells from
patients 1–3 by an average of 2.7-fold above the mean
value of controls. It did not, however, influence total IgG
production by RP105-negative B cells from patient 4.
Similarly, SAC increased total IgM production by
RP105-negative B cells from patients 1 and 3 by an
average of 1.9-fold above the mean value of controls.
Conversely, SAC could not induce IgG from RP105positive B cells, although it induced a small amount of
IgM production by RP105-positive B cells from patient 3
(Figure 2).
Like SAC, rIL-6 (1 ng/ml) increased total IgG
production by RP105-negative B cells from patients 1–3
by an average of 3.1-fold above the mean value of
controls, but exerted no effect on total IgG production
by RP105-negative B cells from patient 4. Similarly,
RESULTS
RP105-negative B cells in SLE patients. The
frequency of peripheral RP105-negative B cells ranged
from 8.8% to 31.0% (mean 19.5%) (Table 1). RP105negative B cells, which are rare in normal subjects,
appeared in significant numbers in SLE patients, as
previously described (20).
IgG and IgM production by RP105-positive and
RP105-negative B cells obtained from SLE patients.
RP105-positive and RP105-negative B cells obtained
from SLE patients with active disease (patients 1–4)
were cultured separately without stimulation, and IgG
and IgM produced in the supernatant were measured on
days 3 and 5. RP105-negative B cells spontaneously
produced IgG and IgM (13.4 ⫾ 5.8 ␮g/ml and 245 ⫾ 74.6
ng/ml, respectively, on day 5), but RP105-positive B cells
Figure 1. Spontaneous production of polyclonal immunoglobulins by
RP105-negative B cells obtained from 4 patients with systemic lupus
erythematosus. Highly purified RP105-positive and RP105-negative B
cells were cultured separately for 5 days. Supernatants were harvested
after 3 and 5 days of culture, and IgG and IgM secretion was measured
by enzyme-linked immunosorbent assay.
3262
Figure 2. The effect of Staphylococcus aureus Cowan 1 (SAC) on
immunoglobulin production in RP105-positive and RP105-negative B
cells obtained from 4 patients with systemic lupus erythematosus.
RP105-positive and RR105-negative B cells were cultured separately
for 5 days in the presence of 0.001% SAC. The culture supernatants
were assessed for total IgG and IgM by enzyme-linked immunosorbent
assay.
rIL-6 enhanced total IgM production by RP105-negative
B cells from patients 1 and 3 by an average 2.0-fold
above the mean value of controls. In RP105-positive B
cells, however, the addition of rIL-6 did not induce
production of either IgG or IgM (Figure 3).
In an attempt to confirm the effect of IL-6,
anti–IL-6 antibody was added in this system. Anti–IL-6
Figure 3. The effect of interleukin-6 (IL-6) or anti–IL-6 antibody on
immunoglobulin production in RP105-positive and RP105-negative B
cells obtained from 4 patients with systemic lupus erythematosus.
RP105-positive and RP105-negative B cells were cultured separately
for 5 days in the presence of IL-6 (1 ng/ml) or anti–IL-6 antibody (50
ng/ml). The culture supernatants were assessed for total IgG and IgM
by enzyme-linked immunosorbent assay.
KIKUCHI ET AL
Figure 4. Spontaneous interleukin-6 (IL-6) production by RP105positive and RP105-negative B cells obtained from patients with
systemic lupus erythematosus. The culture supernatants that were
harvested as described in Figure 1 were measured for the quantity of
IL-6.
antibody inhibited both IgG and IgM production (an
average of 18% and 30%, respectively) by RP105negative B cells from patients 1–4 (Figure 3).
IL-6 production by RP105-positive and RP105negative B cells. Although SAC and IL-6 exerted no
effect on IgG and IgM production by RP105-negative B
cells from patient 4, anti–IL-6 antibody reduced production of both IgG and IgM. This suggested that endogenous production of IL-6 would contribute to spontaneous production of immunoglobulins. Therefore, we
measured IL-6 production in the supernatants of unstimulated cultures, using an IL-6–specific ELISA. It
was demonstrated that both RP105-positive and RP105negative B cells (patients 2–4) produced IL-6. In particular, the concentration of IL-6 produced by RP105negative B cells obtained from patient 4 was extremely
high (Figure 4). These results suggest that a large
amount of endogenous IL-6 production increased production of immunoglobulins, rendering the extra SAC
and IL-6 stimulation ineffective in patient 4. In this
respect, the results suggest that RP105-negative B cells
produce immunoglobulins, with help from autocrine and
paracrine IL-6.
Anti-DNA antibody production by RP105negative B cells obtained from SLE patients. Among
several autoantibodies, anti-DNA antibodies are particularly important in the pathophysiology of SLE (1–4). It
was of great interest, therefore, to determine whether
SIGNIFICANT ROLE OF RP105-NEGATIVE B CELLS IN SLE
these activated and immunoglobulin-producing RP105negative B cells would also be capable of producing
anti-DNA antibodies. We measured the amount of
anti-ssDNA and anti-dsDNA antibodies in the culture
supernatant of RP105-positive and RP105-negative B
cells obtained from patients 1, 3, 5, 6, and 7. In this
system, however, the amount of these specific antibodies
was too small to be compared, although the antibodies
were detected. It has been shown that anti-CD3–
stimulated T cells induce terminal differentiation of B
cells into nondividing high-rate immunoglobulinsecreting plasma cells (25), and that IL-10, present at an
elevated level in the sera of SLE patients, may contribute to the abnormal production of immunoglobulins and
autoantibodies in SLE (26).
This prompted us to culture RP105-positive and
RP105-negative B cells obtained from SLE patients
together with anti-CD3–activated T cells obtained from
normal subjects, in the presence of IL-10 for 10 days. On
day 10, both subsets of B cells were still viable (⬎50%),
using trypan blue dye exclusion. With the help of
activated T cells and IL-10, even RP105-positive B cells
became capable of producing total IgG, although the
amount was apparently small compared with that produced by RP105-negative B cells (mean ⫾ SD 16.0 ⫾
6.5 ␮g/ml versus 56.2 ⫾ 20.6 ␮g/ml) (Figure 5). It was
also observed that a very small amount of anti-ssDNA
3263
Figure 6. Anti–double-stranded DNA (anti-dsDNA) antibody production by RP105-negative B cells. Culture supernatants collected as
described in Figure 5 were assayed for anti–single-stranded DNA
(anti-ssDNA) antibody and anti-dsDNA antibody. Bars show the
mean ⫾ SD.
antibodies was detected in the supernatant of RP105positive B cells obtained from 2 (patients 3 and 5) of 5
SLE patients. In contrast, however, RP105-negative B
cells from all 5 patients produced a significant amount of
anti-ssDNA antibodies (29.4 ⫾ 18.1 AU/ml) (Figure 6).
Most importantly, RP105-negative B cells obtained from
all SLE patients were found to be capable of producing
anti-dsDNA antibodies (0.133 ⫾ 0.107 IU/ml), whereas
RP105-positive B cells from any of the 5 patients did not
produce the antibodies (Figure 6). B cells from normal
subjects did not produce either anti-ssDNA or antidsDNA antibodies (data not shown).
DISCUSSION
Figure 5. IgG production by RP105-positive and RP105-negative B
cells with T cell help. RP105-positive and RP105-negative B cells
(obtained from patients 1, 3, 5–7) were cultured for 10 days with
activated T cells obtained from the normal subject, together with
interleukin-10. The culture supernatants were assayed for total IgG.
Bars show the mean ⫾ SD.
RP105, detected on B cells, has been shown to be
associated with B cell proliferation and resistance to
apoptosis in mice (12). We previously demonstrated that
most human B cells also have RP105 molecules (20).
Interestingly, however, we found that a significant percentage of B cells obtained from patients with SLE,
Sjögren’s syndrome, and dermatomyositis are devoid of
RP105; all of these diseases are known to be characterized by B cell activation (21,22). In particular, in SLE,
the number of RP105-negative B cells was well correlated with disease activity and the serum level of IgG.
Characterization of RP105-negative B cells revealed that
they were IgD-negative and CD38-bright with the expression of CD95 and CD86, being consistent with
activated B cells or germinal center B cells (20). Moreover, RP105-negative B cells had intracellular IgG,
suggesting that RP105-negative B cells can be induced to
produce class-switched immunoglobulin (IgG) (20).
3264
Harada et al reported that early plasma cells, in which
the intensity of expression of CD38 antigen was between
that of germinal center B cells and that of bone marrow
plasma cells, were significantly increased in the peripheral blood of patients with active SLE, bacterial
septicemia, or liver cirrhosis (27). The phenotype of
RP105-negative B cells appears to be similar to that of
the early plasma cells.
In this study, we demonstrated that only a subset
of RP105-negative B cells, which represented ⬃20% of
B cells (Table 1), produced a significant amount of IgG
and IgM by day 3 without any stimulation. It is known
that peripheral B cells from patients with SLE contain
populations that spontaneously produce immunoglobulins and also mature into antibody-secreting cells when
cultured in vitro in the absence of obvious activators of
B cell differentiation (5–7). The ability of these cell
populations to spontaneously produce immunoglobulins
may be equal to or very close to that of RP105-negative
B cells. Conversely, RP105-positive B cells are quite
different in that they did not produce any immunoglobulins, either spontaneously or even with stimulation by
IL-6 or SAC.
IL-6 is a main cytokine that induces terminal
differentiation of B cells into plasma cells and also
protects early plasma cells from apoptosis (28). It is
known that the serum concentration of IL-6 is elevated
in patients with active SLE (29,30). In this study, we
demonstrated that IL-6 enhanced production of both
IgG and IgM by the RP105-negative B cells obtained
from 3 of 4 SLE patients tested, and that this effect of
IL-6 was abrogated by anti–IL-6 antibody. Interestingly,
RP105-negative B cells from patient 4, which did not
respond to exogenous IL-6, produced, by themselves, a
significant amount of IL-6 (Figure 4), suggesting the
existence of IL-6–mediating autocrine B cell activation.
This possibility seems to be supported by Nagafuchi et
al, who demonstrated that constitutive expression of
IL-6 receptors on B cells in conjunction with spontaneous IL-6 production by B cells induces autocrine B cell
activation, which may lead to B cell hyperactivity and
autoantibody secretion in patients with SLE (31,32). In
this respect, immunoglobulin production by RP105negative B cells could be enhanced by the autocrine/
paracrine IL-6 loop. Our result, that RP105-positive B
cells did not respond to IL-6 to produce immunoglobulins despite the ability to produce a certain amount of
IL-6, suggests that those cells may not be differentiated
enough to mediate an IL-6 signal into the cells.
It is now delineated that in patients with SLE, a B
cell subset lacking RP105 molecules can produce auto-
KIKUCHI ET AL
antibodies such as anti-ssDNA and anti-dsDNA antibodies as well as nonspecific total IgG. Our results
suggest that anti-DNA antibodies are disease-specific,
because RP105-negative B cells obtained from patients
with Sjögren’s syndrome or dermatomyositis failed to
produce these antibodies (data not shown). With respect
to autoantibody formation, however, T cell help seems
to be of great importance, because RP105-negative B
cells alone produced a very small amount of antibodies.
In this system, the T cells should only be activated and
are not necessarily required to be autoreactive. In
contrast, RP105-positive B cells were shown to be activated to produce IgG with T cell help, probably by
CD40–CD40 ligand interaction, but failed to produce
anti-dsDNA antibodies. These findings indicate that
RP105-negative B cells obtained from SLE patients are
already activated, and that some of them, if not all, are
committed to differentiate to produce autoantibodies.
In lupus mice, a B cell subset that expresses CD5
is known to produce autoantibodies and to be associated
with the pathogenesis of the disease (33–35), whereas in
other mouse models the autoantibody production is
independent of CD5-positive B cells (36). In humans,
however, CD5-positive B cells are clearly distinct from
RP105-negative B cells and do not seem to be associated
with the pathogenesis of SLE (20,37–40). So far, despite
extensive efforts, any B cell subset that can produce
autoantibodies in SLE has not been identified. To our
knowledge, this may be the first study that identified the
B cell subset (RP105-negative B cells) capable of producing anti-DNA antibodies in SLE. It remains to be
elucidated how autoantibody-producing RP105-negative
B cells emerge in SLE, because many attempts to induce
RP105-negative B cells from RP105-positive cells have
not succeeded yet (data not shown).
In conclusion, besides being a useful marker for
the disease activity of SLE, as previously described (20),
RP105-negative B cells, an activated B cell subset, may
be directly involved in the pathogenesis of SLE through
production of anti-DNA antibodies.
REFERENCES
1. Koffler D, Agnello V, Thoburn R, Kunkel HG. Systemic lupus
erythematosus: prototype of immune complex nephritis in man. J
Exp Med 1971;134 Suppl:169–79.
2. Winfield JB, Faiferman I, Koffler D. Avidity of anti-DNA antibodies in serum and IgG glomerular eluates from patients with
systemic lupus erythematosus: association of high avidity antinative DNA antibody with glomerulonephritis. J Clin Invest 1977;59:
90–6.
3. Brinkman K, Termaat R, Berden JHM, Smeenk RJT. Anti-DNA
SIGNIFICANT ROLE OF RP105-NEGATIVE B CELLS IN SLE
4.
5.
6.
7.
8.
9.
10.
11.
12.
13.
14.
15.
16.
17.
18.
19.
20.
21.
22.
antibodies and lupus nephritis: the complexity of crossreactivity.
Immunol Today 1990;11:232–4.
Pearson L, Lightfoot RW Jr. Correlation of DNA-anti-DNA
association rates with clinical activity in systemic lupus erythematosus (SLE). J Immunol 1981;126:16–9.
Schwab J, Lukowsky A, Volk HD, Peter HH, Melchers I. Precursor frequencies for DNA-specific B lymphocytes in patients with
systemic lupus erythematosus (SLE). Clin Exp Immunol 1994;96:
450–7.
Spronk PE, Horst G, Van Der Gun BT, Limburg PC, Kallenberg
CG. Anti-dsDNA production coincides with concurrent B and T
cell activation during development of active disease in systemic
lupus erythematosus (SLE). Clin Exp Immunol 1996;104:446–53.
Bourne T, Zukowska-Cooper M, Salaman MR, Seifert MH,
Isenberg DA. Spontaneous immunoglobulin-producing capacity of
cultures from lupus patients and normal donors following depletion of cells expressing CD19 or CD38. Clin Exp Immunol
1998;111:611–6.
Dilosa RM, Maeda K, Masuda A, Szakal AK, Tew JG. Germinal
center B cells and antibody production in the bone marrow.
J Immunol 1991;146:4071–7.
Liu YJ, Johnson GD, Gordon J, MacLennan IC. Germinal centers
in T-cell-dependent antibody responses. Immunol Today 1992;13:
17–21.
MacLennan IC, Gulbranson-Judge A, Toellner KM, CasamayorPalleja M, Chan E, Sze DM, et al. The changing preference of T
and B cells for partners as T-dependent antibody responses
develop. Immunol Rev 1997;156:53–66.
Liu YJ, Arpin C. Germinal center development. Immunol Rev
1997;156:111–26.
Miyake K, Yamashita Y, Hitoshi Y, Takatsu K, Kimoto M. Murine
B cell proliferation and protection from apoptosis with an antibody
against a 105-kd molecule: unresponsiveness of X-linked immunodeficient B cells. J Exp Med 1994;180:1217–24.
Miyake K, Yamashita Y, Ogata H, Sudo T, Kimoto M. RP105, a
novel B cell surface molecule implicated in B cell activation, is a
member of the leucine-rich repeat protein family. J Immunol
1995;154:3333–40.
Yamashita Y, Miyake K, Miura Y, Kaneko Y, Yagita H, Suda T,
et al. Activation mediated by RP105 but not CD40 makes normal
B cells susceptible to anti-IgM-induced apoptosis: a role for Fc
receptor coligation. J Exp Med 1996;184:113–20.
Miyake K, Shimazu R, Kondo J, Niki T, Akashi S, Ogata H, et al.
Mouse MD-1, a molecule that is physically associated with RP105
and positively regulates its expression. J Immunol 1998;161:
1348–53.
Ogata H, Su IH, Miyake K, Nagai Y, Akashi S, Mecklenbrauker I,
et al. The toll-like receptor protein RP105 regulates lipopolysaccharide signaling in B cells. J Exp Med 2000;192:23–9.
Miura Y, Miyake K, Yamashita Y, Shimazu R, Copeland NG,
Gilbert DJ, et al. Molecular cloning of a human RP105 homologue
and chromosomal localization of the mouse and human RP105
genes (Ly64 and LY64). Genomics 1996;38:299–304.
Fugier-Vivier I, de Bouteiller O, Guret C, Fossiez F, Banchereau
J, Mattei MG, et al. Molecular cloning of human RP105. Eur
J Immunol 1997;27:1824–7.
Miura Y, Shimazu R, Miyake K, Akashi S, Ogata H, Yamashita Y,
et al. RP105 is associated with MD-1 and transmits an activation
signal in human B cells. Blood 1998;92:2815–22.
Koarada S, Tada Y, Ushiyama O, Morito F, Suzuki N, Ohta A, et
al. B cells lacking RP105, a novel B cell antigen, in systemic lupus
erythematosus. Arthritis Rheum 1999;42:2593–600.
Kikuchi Y, Koarada S, Tada Y, Ushiyama O, Morito F, Suzuki N,
et al. Difference in B cell activation between dermatomyositis and
polymyositis: analysis of the expression of RP105 on peripheral
blood B cells. Ann Rheum Dis 2001;60:1137–40.
Koarada S, Tada Y, Kikuchi Y, Ushiyama O, Suzuki N, Ohta A, et
3265
23.
24.
25.
26.
27.
28.
29.
30.
31.
32.
33.
34.
35.
36.
37.
38.
39.
40.
al. CD180 (RP105) in rheumatic diseases. Rheumatology (Oxford)
2001;40:1315–6.
Tan EM, Cohen AS, Fries JF, Masi AT, McShane DJ, Rothfield
NF, et al. The 1982 revised criteria for the classification of systemic
lupus erythematosus. Arthritis Rheum 1982;25:1271–7.
Bombardier C, Gladman DD, Urowitz MB, Caron D, Chang CH.
Derivation of the SLEDAI: a disease activity index for lupus
patients. The Committee on Prognosis Studies in SLE. Arthritis
Rheum 1992;35:630–40.
Vernino L, McAnally LM, Ramberg J, Lipsky PE. Generation of
nondividing high rate Ig-secreting plasma cells in cultures of
human B cells stimulated with anti-CD3-activated T cells. J Immunol 1992;148:404–10.
Llorente L, Zou W, Levy Y, Richaud-Patin Y, Wijdenes J,
Alcocer-Varela J, et al. Role of interleukin 10 in the B lymphocyte
hyperactivity and autoantibody production of human systemic
lupus erythematosus. J Exp Med 1995;181:839–44.
Harada Y, Kawano MM, Huang N, Mahmoud M, Lisukov I,
Mihara K, et al. Identification of early plasma cells in peripheral
blood and their clinical significance. Br J Haematol 1996;92:
184–91.
Kawano MM, Mihara K, Huang N, Tsujimoto T, Kuramoto A.
Differentiation of early plasma cells on bone marrow stromal cells
requires interleukin-6 for escaping from apoptosis. Blood 1995;85:
487–94.
Linker-Israeli M, Deans RJ, Wallace DJ, Prehn J, Ozeri-chen T,
Klinenberg JR. Elevated levels of endogenous IL-6 in systemic
lupus erythematosus: a putative role in pathogenesis. J Immunol
1991;147:117–23.
Spronk PE, terBorg EJ, Limburg PC, Kallenberg CG. Plasma
concentration of IL-6 in systemic lupus erythematosus; an indicator of disease activity? Clin Exp Immunol 1992;90:106–10.
Nagafuchi H, Suzuki N, Mizushima Y, Sakane T. Constitutive
expression of IL-6 receptors and their role in the excessive B cell
function in patients with systemic lupus erythematosus. J Immunol
1993;151:6525–34.
Kitani A, Hara M, Hirose T, Harigai M, Suzuki K, Kawakami M,
et al. Autostimulatory effects of IL-6 on excessive B cell differentiation in patients with systemic lupus erythematosus: analysis of
IL-6 production and IL-6R expression. Clin Exp Immunol 1992;
88:75–83.
Casali P, Notkins AL. CD5⫹ B lymphocytes, polyreactive antibodies and the human B-cell repertoire. Immunol Today 1989;10:
364–8.
Kocks C, Rajewsky K. Stable expression and somatic hypermutation of antibody V regions in B-cell developmental pathways.
Annu Rev Immunol 1989;7:537–59.
Hayakawa K, Hardy RR, Parks DR, Herzenberg LA. The “Ly-1
B” cell subpopulation in normal, immunodefective, and autoimmune mice. J Exp Med 1983;157:202–18.
Reap EA, Sobel ES, Cohen PL, Eisenberg RA. Conventional B
cells, not B-1 cells, are responsible for producing autoantibodies in
lpr mice. J Exp Med 1993;177:69–78.
Suzuki N, Sakane T, Engleman EG. Anti-DNA antibody production by CD5⫹ and CD5⫺ B cells of patients with systemic lupus
erythematosus. J Clin Invest 1990;85:238–47.
Hayakawa K, Hardy RR. Normal, autoimmune, and malignant
CD5⫹ B cells: the Ly-1 B lineage? Annu Rev Immunol 1988;6:
197–218.
Hayakawa K, Hardy RR, Honda M, Herzenberg LA, Steinberg
AD, Herzenberg LA. Ly-1 B cells: functionally distinct lymphocytes that secrete IgM autoantibodies. Proc Natl Acad Sci U S A
1984;81:2494–8.
Casali P, Burastero SE, Balow JE, Notkins AL. High-afinity
antibodies to ssDNA are produced by CD-B cells in systemic lupus
erythematosus patients. J Immunol 1989;143:3476–83.
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