вход по аккаунту


Development of spontaneous multisystem autoimmune disease and hypersensitivity to antibody-induced inflammation in Fc╨Ю╤Ц receptor IIatransgenic mice.

код для вставкиСкачать
Vol. 52, No. 10, October 2005, pp 3220–3229
DOI 10.1002/art.21344
© 2005, American College of Rheumatology
Development of Spontaneous Multisystem Autoimmune Disease
and Hypersensitivity to Antibody-Induced Inflammation
in Fc␥ Receptor IIa–Transgenic Mice
Caroline Tan Sardjono,1 Patricia L. Mottram,1 Nicholas C. van de Velde,1 Maree S. Powell,1
David Power,1 Ronald F. Slocombe,2 Ian P. Wicks,3 Ian K. Campbell,3 Steven E. McKenzie,4
Mark Brooks,5 Andrew W. Stevenson,6 and P. Mark Hogarth1
transgenic C57BL/6 (H-2b) mice did not develop CIA
when similarly immunized. Passive transfer of a single
dose of anti-CII antibody induced a more rapid, severe
arthritis in Fc␥RIIa-transgenic mice than in nontransgenic animals. In addition, most immune complex–
induced production of tumor necrosis factor ␣ by activated macrophages occurred via Fc␥RIIa, not the
endogenous mouse FcR. A spontaneous, multisystem
autoimmune disease developed in aging (>20 weeks)
transgenic mice (n ⴝ 25), with a 32% incidence of
arthritis, and by 45 weeks, all mice had developed
glomerulonephritis and pneumonitis, and most had
antihistone antibodies. Elevated IgG2a levels were seen
in mice with CIA and in those with spontaneous disease.
Conclusion. The presence of enhanced passive
and induced autoimmunity, as well as the emergence of
spontaneous autoimmune disease at 20–45 weeks of age,
suggest that Fc␥RIIa is a very important factor in the
pathogenesis of autoimmune inflammation and a possible target for therapeutic intervention.
Objective. The major human Fc receptor,
Fc␥RIIa, is the most widespread activating FcR. Our
aim was to determine the role of Fc␥RIIa in a transgenic
mouse model of immune complex–mediated autoimmunity and to characterize the development of spontaneous
autoimmune disease.
Methods. Arthritis was induced in normal and
Fc␥RIIa-transgenic mice by immunization with type II
collagen (CII) or by transfer of arthritogenic anti-CII
antibodies. Also, mice that spontaneously developed
autoimmune disease were assessed by clinical scoring of
affected limbs, histology and serology, and measurement of autoantibody titers and cytokine production.
Results. Fc␥RIIa-transgenic mice developed
collagen-induced arthritis (CIA) more rapidly than did
archetypal CIA-sensitive DBA/1 (H-2q) mice, while nonSupported by grants from the National Health and Medical
Research Council and PrimaBiomed Ltd., Australia. Dr. Sardjono’s
work was supported by PaperlinX Pty Ltd. Drs. Mottram and Powell’s
work was supported by Nancy Prendergast fellowships from the
Arthritis Foundation, Australia.
Caroline Tan Sardjono, PhD, Patricia L. Mottram, PhD,
Nicholas C. van de Velde, BSc Hons, Maree S. Powell, PhD, David
Power, PhD, P. Mark Hogarth, PhD: Austin Research Institute,
Heidelberg, Victoria, Australia; 2Ronald F. Slocombe, PhD: University of Melbourne, Melbourne, Victoria, Australia; 3Ian P. Wicks,
PhD, Ian K. Campbell, PhD: Walter and Eliza Hall Institute, Parkville,
Victoria, Australia; 4Steven E. McKenzie, PhD: Jefferson Medical
College, Philadelphia, Pennsylvania; 5Mark Brooks, MBBS: Austin
Hospital, Heidelberg, Victoria, Australia; 6Andrew W. Stevenson,
PhD: Commonwealth Scientific Industrial Research Organization,
Clayton South, Victoria, Australia.
Drs. Mottram, Powell, and Hogarth have stock options in
PrimaBiomed. Dr. McKenzie has received consulting fees (less than
$10,000 per year) from GlaxoSmithKline.
Address correspondence and reprint requests to P. Mark
Hogarth, PhD, Helen McPherson-Smith Laboratory, Austin Research
Institute, Studley Road, Heidelberg, Victoria 3084, Australia. E-mail:
[email protected]
Submitted for publication September 24, 2004; accepted in
revised form June 30, 2005.
Antibody-induced inflammation is a major component of several autoimmune diseases (1,2). The role
of cell surface receptors for antibodies, especially IgG
Fc␥ receptors (Fc␥R), was recognized following amelioration of tissue destruction in type III hypersensitivity
reactions after administration of soluble recombinant
human Fc␥RIIa in vivo (3). Subsequent studies with
FcR-deficient mice (4) showed that Fc␥R play significant roles in antibody-induced inflammatory disease
models such as collagen-induced arthritis (CIA) (5),
passive antibody-induced arthritis (6), and intraarticular
antigen-induced arthritis (7). However, rodents lack an
ortholog of Fc␥RIIa, the most abundant and widespread
activating FcR in higher primates. Fc␥RIIa has unique
structural, signaling, and biologic features (8–11). Unlike other FcR, Fc␥RIIa can signal without the homodimeric FcR ␥-chain used by Fc␧RI, Fc␥RI, Fc␥RIII,
and Fc␣RI, since both the ligand binding site and the
immunoreceptor tyrosine-based activation motif
(ITAM) are in the same polypeptide (8). Moreover,
Fc␥RIIa is a dimer, with the ITAM-containing cytoplasmic tails arranged in an FcR ␥-chain–like configuration (9,12).
Studies of Fc␥RIIa, as well as other FcR, transfected into mouse or primate cells show that these FcR
behave identically in both ligand binding and activation/
regulation (13,14). The interaction of Fc␥RII ITAMs
and immunoreceptor tyrosine-based inhibition motifs
(ITIMs) was seen in both transfected mouse and human
cell lines, and ITIM sequences in mice and humans are
highly conserved (15). Fc␥RIIa in transgenic mice is
expressed under its own promoter and has the same
expression pattern in mice and humans (16,17). Thus,
Fc␥RIIa can interact appropriately with intracellular
signaling pathways in mouse cells. Finally, genetic polymorphisms of Fc␥RIIa are associated with human autoimmune disease (1,18). In this study, we analyzed inflammatory responses in transgenic mice expressing
Fc␥RIIa and confirmed a role for this receptor in
passive, induced, and spontaneous autoimmune disease.
Mice. DBA/1 (H-2q), C57BL/6 (H-2b), SJL/J (H-2s),
(SJL ⫻ C57BL/6)F1 (H-2b/s), and Fc␥RIIa-transgenic mice
(H-2b) derived from (SJL ⫻ C57BL/6)F2 embryos (17) were
used. The Fc␥RIIa-transgenic mice were inbred for ⬎20
generations and were homozygous for the transgene under the
control of its own promoter. They carried the high responder
(Arg134) allele of Fc␥RIIa, which binds mouse IgG2a, Ig2b,
and Ig1, as well as human IgG1, IgG2, and IgG3 (11).
Induction of CIA. Mice were injected intradermally at
the base of the tail with 100 ␮l of 2 mg/ml type II collagen (CII)
emulsion in Freund’s complete adjuvant (Difco, Detroit, MI)
that contained 2.5 mg/ml heat-killed Mycobacterium tuberculosis H37Ra (Difco). Mice were immunized a second time 21
days later (19). They were examined daily from days 1–60, and
arthritis in each limb was graded on a scale of 0–3 (0 ⫽ normal,
1 ⫽ mild swelling and redness, 2 ⫽ severe swelling/redness,
and 3 ⫽ severe swelling and redness and joint rigidity). The
maximum possible score (arthritis index) was 12 for each
Passively induced arthritis. Anti-CII monoclonal antibody (mAb) M2139 (2 mg) (20) was injected intraperitoneally
into Fc␥RIIa-transgenic and nontransgenic C57BL/6 mice, and
arthritis progression was monitored daily as described above.
Joint histology. Joints were preserved in 10% formalin/
phosphate buffered saline (PBS), decalcified in 5% HCl, 3.5%
glacial acetic acid, 95% ethanol, and 12.5% (volume/volume)
chloroform, and then were embedded in paraffin. Sections
(4–6 ␮m) were stained with hematoxylin and eosin (H&E).
Enzyme-linked immunosorbent assay (ELISA) for
anti-CII antibody. ELISA plates (96-well; Costar, Cambridge,
MA) were coated with 50 ␮g/ml CII and blocked with 2%
bovine serum albumin in PBS (1 hour at room temperature).
Sera (serially diluted) were added, and antibody was detected
using secondary horseradish peroxidase (HRP)–conjugated
sheep anti-mouse IgG F(ab⬘)2 fragments (Amersham, Little
Chalfont, UK). Development was performed for 10 minutes
with ABTS (Boehringer Mannheim, Rockville, MD), and
absorbance was read at 405 nm. IgG isotypes were detected
with specific antibodies (see below).
Bone marrow macrophage cultures. Bone marrow
macrophages were cultured in Dulbecco’s modified Eagle’s
medium (DMEM) (CSL, Melbourne, Victoria, Australia) containing 2 mM glutamine, 50 mM 2-mercaptoethanol, 100
units/ml penicillin, 100 ␮g/ml streptomycin, 10% (v/v) fetal calf
serum, and 30% (v/v) L cell–conditioned medium at 37°C in
10% CO2 for 5–7 days (21).
Peritoneal exudate macrophage (PEM) cultures. Mice
were injected intraperitoneally with 0.5 ml of 4% (weight/
volume) thioglycolate (CSL) 4 days prior to harvest. PEMs
were cultured in DMEM, prepared as described above, for 1–3
Measurement of cytokine production. PEMs (1 ⫻
106/ml) were incubated with heat-aggregated IgG (HAGG; 50
␮g or 100 ␮g) or phorbol myristate acetate (PMA; 20 ng)
(Sigma, St. Louis, MO) at 37°C for 24 hours. HAGG was
prepared from 8–10 mg/ml of human gamma globulin (Sandoglobulin; Novartis, East Hanover, NJ) heated for 30 minutes
at 63°C, then put on ice and brought to 1% (w/v) with
polyethylene glycol 6000 (Sigma) in PBS and kept on ice for 30
minutes. The precipitated complexes were centrifuged
(10,000g at 4°C for 10 minutes), the supernatant was discarded,
and complexes were dissolved in PBS at 1 mg/ml.
Production of tumor necrosis factor ␣ (TNF␣) and
interleukin-10 (IL-10) by macrophages in the supernatant 2–24
hours after stimulation with HAGG or PMA was detected by
ELISA using cytokine detection kits from eBioscience (San
Diego, CA). The specificity of induced TNF␣ secretion was
determined by preincubation of macrophages with antiFc␥RIIa mAb IV.3 Fab (20 or 50 ␮g) at 4°C for 1 hour.
Detection of Fc␥R by flow cytofluorometry. The following Fc␥R were detected: mouse Fc␥RI using mAb X54-5/
7.1 (mIgG1) (22), mouse Fc␥RIIb using mAb Ly-17.2 (mIgG2;
Cedarlane Laboratories, Hornby, Ontario, Canada), and
hFc␥RIIa using mAb IV.3 (mIgG2b) or mAb 8.7 (mIgG1).
Monitoring of spontaneous autoimmunity. Mice were
examined from 12 weeks of age, and the index of arthritis
severity was assessed as described above. Organs were fixed in
10% formalin/PBS, embedded in paraffin, and histologic sections were stained with H&E. Immune complex deposition in
kidney sections was detected with fluorescein isothiocyanate
(FITC)–conjugated sheep anti-mouse IgG (Silenus Laboratories, Hawthorn, Victoria, Australia).
Transmission electron microscopy. Mouse kidneys
were cut into 1–2-mm cubes and fixed in 2–8% paraformalde-
hyde/2–5% glutaraldehyde in 0.15M cacodylate buffer (pH 7.4)
for ⬎6 hours at 4°C. Tissues were rinsed in cacodylate buffer and
postfixed in 1% osmium tetroxide in 0.15M cacodylate buffer (pH
7.4) for 2 hours at room temperature. Samples were washed in
distilled water, dehydrated in 10% incremental concentrations of
acetone, and then embedded in Procure–Araldite resin (ProSci
Tech, Thuringowa, Queensland, Australia). During the dehydration, tissues were stained with 2% uranyl acetate in 70% acetone.
Ultrathin sections were stained with 5% uranyl acetate in aqueous
solution for 30 minutes at room temperature and then were
stained with Reynolds lead citrate for 10 minutes. Sections were
examined using a Philips 300 electron microscope (Philips, Mahwah, NJ) at 60 kV.
Radiographic analysis. Polychromatic hard radiography was performed (23) on mice, and images were recorded
using an FXE-225.20 microfocus x-ray source (Feinfocus,
Stamford, CT) fitted with a tungsten target, operated at 60
kVp. Projection geometry with a large sample-to-detector
distance resulted in high-resolution images. Exposures were
⬃3 mA, and images were recorded with imaging plates (XRT,
Port Melbourne, Victoria, Australia) scanned with a Fuji
BAS-5000 scanner (Fuji Photo Film, Tokyo, Japan).
Detection of antihistone antibodies. ELISA plates
were coated with 20 ␮g/ml purified histone (a mixture of H1,
H2A, H2B, H3, and H4 from calf thymus) (Roche Laboratories, Basel, Switzerland). Serially diluted samples (50 ␮l) were
added, incubated for 1 hour at room temperature, detected
with HRP-conjugated sheep anti-mouse IgG F(ab⬘)2 fragments
(Amersham), and then developed using ABTS.
Quantitation of IgG subclasses. Total serum IgG
concentrations were determined using ELISA plates coated
with 50 ␮l (3 ␮g/ml) of rat anti-mouse IgG (BD PharMingen,
San Diego, CA). Serially diluted serum samples (50 ␮l) were
added and incubated for 1 hour at room temperature. Antibody was detected with biotin-conjugated, isotype-specific rat
anti-mouse IgG1, anti-mouse IgG2a, and anti-mouse IgG2b
mAb (BD PharMingen) (1 hour at room temperature), then
with streptavidin–HRP (1 hour at room temperature), and
developed with ABTS. Quantitation of IgG was by comparison
with class-specific IgG standards.
Statistical analysis. Results were expressed as the
mean ⫾ SD. Statistical differences were analyzed using the
Mann-Whitney 2-sample rank test, correlation coefficients, or
Student’s t-test. All the statistical analyses were performed
using Microsoft Excel software (Microsoft, Redmond, WA). P
values less than 0.05 were considered statistically significant.
Expression of transgenic and endogenous FcR.
The expression of Fc␥R on bone marrow macrophages
from Fc␥RIIa-transgenic and nontransgenic C57BL/6
mice was analyzed by flow cytometry. Using mAb IV.3
(anti-Fc␥RIIa) (Figure 1A), Fc␥RIIa was detected on
macrophages from Fc␥RIIa-transgenic mice but not on
macrophages from nontransgenic mice. The expression
Figure 1. Flow cytometric analysis of Fc␥ receptor (Fc␥R) expression
in normal and human Fc␥RIIa–transgenic mice. Bone marrow macrophages from Fc␥RIIa-transgenic mice (solid histograms) or nontransgenic C57BL/6 mice (dotted line, open histograms) were stained for A,
Fc␥RIIa with monoclonal antibody (mAb) IV.3, B, Fc␥RI with mAb
X54-5/7.1, and C, Fc␥RIIb with mAb Ly-17.2. Studies were performed
with fluorescein isothiocyanate–conjugated anti-mouse IgG (solid line,
open histogram). FL-1 ⫽ fluorescence channel 1.
of the transgene did not significantly change the expression of endogenous mouse FcR, since similar levels of
Fc␥RI (Figure 1B) and especially inhibitory Fc␥RIIb
(Figure 1C) were observed on Fc␥RIIa-transgenic and
nontransgenic macrophages. Similarly, expression of
Fc␥RIIa on neutrophils did not alter endogenous FcR
expression. Fc␥RIIa was not expressed on B or T cells
(data not shown), as expected (16,17).
Fc␥RIIa confers susceptibility to CIA. In mice,
CIA susceptibility is associated with the major histocompatibility complex (MHC) genotype: H-2q and H-2r mice
are highly susceptible, while H-2b, H-2d, and H-2s mice
are less so (19,24). We compared CIA development in
Fc␥RIIa-transgenic (H-2b) mice with the archetypal
susceptible strain DBA/1 (H-2q) and the less susceptible
C57BL/6 (H-2b) and (SJL ⫻ C57BL/6)F1 (H-2s/b) strains
Figure 2. Analysis of collagen-induced arthritis (CIA) in Fc␥RIIatransgenic mice. A, Development of CIA in Fc␥RIIa-transgenic mice
(n ⫽ 41) compared with nontransgenic DBA/1 mice (n ⫽ 27), C57BL/6
mice (n ⫽ 28), and (SJL ⫻ B6)F1 mice (n ⫽ 8). Pooled data from 5
experiments are shown. B, Effects of treatment of arthritis with
anti-Fc␥RIIa F(ab⬘)2 mAb 8.7 (intraperitoneal injection of 100 ␮g on
days 21, 24, 27, and 30 versus treatment with phosphate buffered saline
[PBS] alone [n ⫽ 8 mice per group]). The mAb 8.7 treatment caused
a significant reduction in disease severity on day 40 (P ⬍ 0.05). Values
in A and B are the mean ⫾ SD. C, A representative section showing
histopathologic features of an Fc␥RIIa-transgenic mouse knee joint 36
days after CIA induction (hematoxylin and eosin stained; original
magnification ⫻ 100). D, Histopathologic features of a knee joint from
a C57BL/6 mouse 36 days after collagen injection (hematoxylin and
eosin stained; original magnification ⫻ 40). See Figure 1 for other
(Figure 2A). The Fc␥RIIa-transgenic mice in this experiment were highly susceptible, with more rapid onset of
arthritis (day 18) compared with other mice (days 22 and
24). Of the Fc␥RIIa-transgenic mice, 15% developed
arthritis after 1 immunization, whereas 2 immunizations
were always required for CIA development in DBA/1
and (SJL ⫻ C57BL/6)F1 mice. No arthritis occurred in
C57BL/6 mice under these conditions. By day 26, ⬎90%
of the Fc␥RIIa-transgenic mice developed arthritis,
compared with ⬍10% of DBA/1 and (SJL ⫻ C57BL/
6)F1 mice (P ⬍ 0.001).
Disease incidence and severity at ⬎30 days were
similar in Fc␥RIIa-transgenic and DBA/1 mice. In
Fc␥RIIa-transgenic mice treated with a limited course of
anti-Fc␥RIIa F(ab⬘)2 antibody (100 ␮g/mouse intraperitoneally on days 21, 24, 27, and 30), CIA was significantly reduced on day 35 (P ⬍ 0.05) compared with
untreated CIA in Fc␥RIIa-transgenic mice. Although
this dose of antibody is unlikely to have blocked all in
vivo activity of Fc␥RIIa, the data show that this FcR
plays a role in disease severity in these mice (Figure 2B).
Histologic assessment of paws and ankle and
knee joints from Fc␥RIIa-transgenic mice on day 36 after
arthritis induction (when the maximal clinical index was
observed) showed severe, destructive arthritis with pannus formation, infiltration of inflammatory cells (polymorphonuclear cells and macrophages) into the synovial
space, and erosion of the cartilage (Figure 2C). No joint
inflammation was seen in C57BL/6 controls (Figure 2D).
Correlation of increased levels of IgG2a with the
arthritis index in transgenic mice. Despite the accelerated development of severe CIA, Fc␥RIIa-transgenic
mice had lower anticollagen antibody titers than susceptible DBA/1 mice, but similar to those of the less
susceptible (SJL ⫻ C57BL/6)F1 mice (Figure 3A). The
anticollagen antibodies were predominantly IgG2, and
analysis of the serum IgG2 subclasses (Figure 3B)
showed a significant increase in the IgG2a:IgG2b ratio
in severely arthritic mice (P ⬍ 0.02) compared with
mildly arthritic or unaffected Fc␥RIIa-transgenic mice.
There was a positive correlation between the arthritis
index and IgG2a levels (r ⫽ 0.57) and no correlation
between arthritis index and IgG2b levels (r ⫽ ⫺0.16).
This suggests a dominant Th1 response in Fc␥RIIatransgenic mice following induction of CIA and is consistent with previous data obtained from DBA/1 mice
(25). Thus, expression of Fc␥RIIa, which binds mouse
IgG2a avidly, may cause effector cells in the Fc␥RIIatransgenic mice to be sensitive to low levels of
autoantibody/immune complex activation, possibly triggering the early release of inflammatory mediators.
Production of TNF␣ by IgG-stimulated macrophages from Fc␥RIIa-transgenic mice. Because TNF␣ is
a major inflammatory mediator in human RA and
mouse CIA (24,26,27), we compared the production of
TNF␣ from immune complex–stimulated Fc␥RIIatransgenic and nontransgenic macrophages. HAGGstimulated Fc␥RIIa macrophages produced significantly
more TNF␣ (6.5 ng/ml) compared with nontransgenic
macrophages (1 ng/ml; P ⫽ 0.001) (Figure 3C). Elevated
TNF␣ production by Fc␥RIIa macrophages was blocked
(although not completely) in a dose-dependent manner
by anti-Fc␥RIIa mAb IV.3 Fab to levels not significantly
different from those produced by nontransgenic macrophages, indicating that immune complex–induced TNF␣
production was due principally to activation by Fc␥RIIa,
rather than through the endogenous mouse activating
Fc␥R. PMA stimulation of nontransgenic and transgenic
macrophages showed that these cells were equally responsive (Figure 3C). No difference in IL-10 production
Figure 3. Sensitivity of human Fc␥ receptor IIa (Fc␥RIIa)–transgenic
mice to antibody-mediated disease. A, Anticollagen antibody titers in
Fc␥RIIa-transgenic (Tg) and nontransgenic (SJL ⫻ C57BL/6)F1 and
DBA/1 mice (n ⫽ 5 per group), as determined on days 0, 21, and 36
postimmunization. OD ⫽ optical density. B, Comparison of changes in
IgG2 subclasses in Fc␥RIIa-transgenic mice without collagen-induced
arthritis (CIA) (index 0), with severe arthritis (index 5–12; high CIA),
and with mild arthritis (index 1–4; low CIA) after the second injection
of collagen (n ⫽ 5 mice per group). In the high CIA group, there was
a significant increase in the IgG2a:IgG2b ratio (P ⫽ 0.016 versus mildly
arthritic mice, by Student’s t-test), a positive correlation between the
arthritis index and IgG2a levels (r ⫽ 0.57), and no correlation between
the arthritis index and IgG2b levels (r ⫽ ⫺0.16). C, Comparison of
tumor necrosis factor ␣ (TNF␣) production in heat-aggregated IgG
(HAGG)–stimulated macrophages. Macrophages from Fc␥RIIatransgenic mice produced significantly higher levels of TNF␣ compared with nontransgenic C57BL/6 (B6) mice (P ⫽ 0.001) following
stimulation with HAGG. There was no significant difference (P ⫽
0.07) between Fc␥RIIa-transgenic and C57BL/6 mouse macrophages
treated with HAGG following incubation with 20- or 50-␮g doses of
monoclonal antibody (mAb) IV.3. Responses to phorbol myristate
acetate (PMA) stimulation were equivalent in both strains of macrophages (P ⬎ 0.05). D, Arthritis index over time in Fc␥RIIa-transgenic
and (SJL ⫻ C57BL/6)F1 mice treated with 2 mg of mAb M2139 given
intraperitoneally on day 0 (n ⫽ 6 per group). Fc␥RIIa-transgenic mice
treated with mAb M2139 were hyperresponsive to anti–type II collagen. Values are the mean and SD.
was observed in HAGG-stimulated macrophages (data
not shown).
Exaggerated antibody hypersensitivity of
Fc␥RIIa-transgenic mice. The rapid CIA response in
Fc␥RIIa-transgenic mice and increased sensitivity to
HAGG implied an exaggerated response to pathologic
antibodies. This possibility was tested using a passive
antibody-transfer arthritis model described by Holmdahl
and colleagues (20,28), wherein mice were normally
given a single dose of a mixture of 2 anticollagen
antibodies (4.5 mg each of M2139 and C1), followed by
50 ␮g of lipopolysaccharide (LPS) 5 days later. These
antibodies have been tested for arthritis induction in a
number of mouse strains, including SJL, SJL ⫻ B6,
BALB/c, and DBA/1 (28). None of these strains responded without LPS. However, in Fc␥RIIa-transgenic
mice (Figure 3D), treatment with a single 2-mg dose of
mAb M2139 alone, without LPS, caused a rapid onset of
arthritis in 100% of mice, but had no effect in nontransgenic controls.
Spontaneous development of autoimmunity in
Fc␥RIIa-transgenic mice. Mice were housed for ⬎1
year, and we observed spontaneous progressive development of a systemic multiorgan autoimmune syndrome.
Development of destructive arthritis. A proportion
of aging Fc␥RIIa-transgenic mice spontaneously developed severe, destructive, symmetric arthritis. In a group
of 25 Fc␥RIIa-transgenic mice monitored for ⬎1 year,
none younger than 20 weeks of age developed arthritis,
7 (28%) developed arthritis between 20–45 weeks of
Figure 4. Analysis of arthritis in mice with spontaneous autoimmune
disease. A, Cumulative percentage incidence in arthritis over time in
human Fc␥ receptor IIa (Fc␥RIIa)–transgenic mice (n ⫽ 25). There
was no difference in arthritis incidence between males and females. B
and C, Photograph and radiographic image of the paw of a 36-weekold Fc␥RIIa-transgenic mouse with severe spontaneous arthritis (B)
and a nonarthritic Fc␥RIIa-transgenic age-matched control mouse
(C). Radiography was performed for each mouse category, i.e.,
nontransgenic, healthy Fc␥RIIa-transgenic, Fc␥RIIa-transgenic with
mild arthritis, and Fc␥RIIa-transgenic with destructive arthritis (n ⫽ 4
per group) (results not shown). D–F, Representative sections showing
the histopathologic features of hematoxylin and eosin–stained mouse
joints. D, Early active destructive arthritis in an ankle from a 28-weekold mouse, with polymorphonuclear-dominant inflammatory cell infiltration. E, A later stage of destructive disease in a knee from a
36-week-old mouse, with predominantly mononuclear cell infiltration
and advanced pannus. F, A normal ankle joint from an older (age 36
weeks) transgenic mouse. ca ⫽ cartilage; p ⫽ pannus; i ⫽ inflammatory cells. (Original magnification ⫻ 100.)
age, and 1 of the 25 developed arthritis thereafter, with
a cumulative incidence of 32% (Figure 4A). Of the 8
mice with arthritis, 5 were severely affected (mean ⫾ SD
index 10 ⫾ 2.38), with profoundly swollen joints and
severe histologic and radiologic changes (Figure 4). The
other 3 Fc␥RIIa-transgenic mice had less severe arthritis. Despite differences in arthritis severity, in both
subsets of Fc␥RIIa-transgenic mice, the number of
affected paws was similar. Age-matched nontransgenic
mice never developed spontaneous disease.
Radiologic examination of the affected limbs. The
Fc␥RIIa-transgenic mice with severe arthritis developed
marked radiologic changes that mirrored the florid
edema and distortion of the foot seen clinically. Severe
ankylosis of the tibiotarsal and tarsophalangeal cartilage
was seen, with loss of joint space and prominent periarticular new bone formation (Figure 4B) compared with
age-matched Fc␥RIIa-transgenic mice that did not develop arthritis (Figure 4C). Limbs from mice with mild
arthritis revealed minimal radiologic changes (results
not shown).
Histologic evaluation of the joints. Joint histology
of Fc␥RIIa-transgenic mice compared with that of nontransgenic, age-matched controls (Figures 4D–F)
showed that mice with spontaneous severe arthritis had
synovial hyperplasia and proliferation, cartilage erosion,
pannus formation, and joint space infiltrate. The later
stage of the disease showed more advanced destruction
of bone and thinning of the cartilage, with the infiltrate
changing from polymorphonuclear (PMN) cells in early
active disease (Figure 4D) to macrophages in more
advanced cases (Figure 4E). There was no evidence of
disease in age-matched C57BL/6 or (SJL ⫻ C57BL/6)F1
controls (Figure 4F).
Serum IgG2a elevation in affected mice. The levels
of total IgG, IgG1, IgG2a, IgG2b, and IgG3 in all mice
were determined by ELISA. Only the total level of
IgG2a antibody was elevated (Figure 5A), and only in
Fc␥RIIa-transgenic mice that developed destructive arthritis (mean ⫾ SD 6.6 ⫾ 2.5 ␮g/ml). IgG2a levels in
mice with mild arthritis (2.0 ⫾ 0.8 ␮g/ml) were similar to
those in Fc␥RIIa-transgenic mice with no disease (2.6 ⫾
1.2 ␮g/ml) and in nontransgenic mice (1.7 ⫾ 1.3 ␮g/ml).
Thus, there was a strong correlation between IgG2a
levels and disease severity, particularly with the formation of pannus (r ⫽ 0.63). The levels of the other IgG
subclasses were not altered (Figure 5).
Other features of systemic autoimmune disease
in Fc␥RIIa-transgenic mice. The Fc␥RIIa-transgenic
mice were screened for further evidence of inflammation and autoimmunity by histologic examination of
Figure 5. Serologic findings and incidence of kidney and lung disease
in mice with spontaneous autoimmune disease. A, Levels of serum
IgG2a in individual human Fc␥ receptor IIa (FcR␥IIa)–transgenic
mice with destructive arthritis (n ⫽ 8) compared with unaffected
FcR␥IIa-transgenic mice (n ⫽ 7), FcR␥IIa-transgenic mice with mild
arthritis (n ⫽ 6), and normal age-matched nontransgenic mice (n ⫽ 6).
Bars indicate the mean ⫾ SD concentration of IgG2a in each group. P
⫽ 0.0012 for IgG2a levels in mice with destructive disease versus
unaffected nontransgenic mice. IgG2a levels correlated with disease
severity, particularly with the formation of pannus (r ⫽ 0.63). Total
IgG was increased in arthritic mice, and this correlated strongly with
IgG2a levels (r ⫽ 0.96). The levels of the other Ig subclasses (IgM,
IgG1, Ig2b, and Ig3) were not significantly different between the 4
groups of mice (P ⬎ 0.05 for all comparisons). B, Incidence of
glomerulonephritis (GN) and pneumonitis (Pn) in Fc␥RIIa-transgenic
mice at ages ⬍20 weeks (n ⫽ 8), 21–40 weeks (n ⫽ 14), and ⬎40 weeks
(n ⫽ 9).
H&E-stained sections of skin, lymph nodes, gut, salivary
glands, kidneys, eyes, brain, lungs, spleen, liver, pancreas, and heart at 14–60 weeks of age and compared
with aged-matched, nontransgenic controls. Glomerulonephritis (GN) and pneumonitis were commonly observed in Fc␥RIIa-transgenic mice, with the disease
incidence increasing with age. By 50 weeks of age, all
mice were affected (Figure 5B). No abnormalities were
found in other organs or in nontransgenic mice of any
Glomerulonephritis. The time of onset and degree
of severity of GN in individuals varied considerably. Few
mice developed GN before 20 weeks of age, and up to
80% had the disease by 40 weeks. However, all mice ⬎40
weeks of age had moderate to severe GN, implying an
age-related progression to severe GN (Figure 5B). Multifocal lymphoplasmacytic infiltrate in the renal interstitium, mainly around major arcuate vessels, was seen at
25–30 weeks (Figure 6A), with mild mesangial matrix
deposition in the glomeruli and some tubular thickening.
In mice ages ⬎40 weeks, more advanced disease was
seen, with enlarged glomeruli, increased mesangial matrix (Figures 6B and C), and condensation of glomerular
tufts. There were proliferative and sclerotic changes in
Bowman’s capsule, indicative of crescent formation, and
mild tubular proliferative changes (Figure 6B), although
tubulointerstitial infiltrates were not present. The dis-
Figure 6. Analysis of spontaneous autoimmune glomerulonephritis,
pneumonitis, and antihistone antibodies in a group of older human
Fc␥ receptor IIa (Fc␥RIIa)–transgenic mice (n ⫽ 31). A, Kidney
section (hematoxylin and eosin [H&E] stained) taken at age 25 weeks,
showing multifocal lymphoplasmacytic infiltration in the renal interstitium, mainly around major arcuate vessels. The glomerulus (arrow)
shows mild mesangial matrix deposition. Tubules were thickened but
otherwise normal (original magnification ⫻ 200). B, An H&E-stained
kidney section taken at 45 weeks, showing enlarged glomeruli (thin
arrows), increased mesangial matrix, and condensation of glomerular
tufts. There are proliferative and sclerotic changes in Bowman’s
capsule (thick arrow) indicative of crescent formation, and mild
tubular proliferative changes (original magnification ⫻ 100). C, Fluorescence staining of immune complexes in the glomerulus. Kidney
sections were stained with anti-mouse IgG, directly conjugated with
fluorescein isothiocyanate, and immune complexes within the glomerulus appeared granular (original magnification ⫻ 200). Diffuse, lowlevel staining with no glomerular concentration of IgG was seen in
age-matched, nontransgenic control mice (results not shown). D,
Transmission electron microscopy of glomeruli, showing immune
complexes deposited (D) on the glomerular basement membrane (BM,
arrow) above the uriniferous space (U) and below the endothelial layer
(En) (original magnification ⫻ 50). E, H&E-stained Fc␥RIIatransgenic mouse lung section taken at 40 weeks, showing infiltration
of inflammatory cells (arrows) and local destruction of lung architecture (original magnification ⫻ 50). F, Enzyme-linked immunosorbent
assay for the presence of antihistone antibodies in Fc␥RIIa-transgenic
and nontransgenic mouse sera at 36 weeks of age. Antihistone
antibodies were present in serum from many of the Fc␥RIIatransgenic mice at all ages, but were not present in serum from the
nontransgenic mice. Horizontal line shows the cutoff for positivity.
ease was self-limiting, since mice ages ⬎40 weeks, with
up to 65% of glomeruli affected, remained healthy, with
normal urinary protein and serum creatinine levels
(results not shown). Age-matched nontransgenic mice
showed no evidence of disease (results not shown).
FITC-conjugated anti-mouse IgG staining of kidney sections showed an accumulation of immune complexes within the glomeruli, which produced a dense,
granular appearance (Figure 6C). In contrast, kidneys
from age-matched nontransgenic control mice showed
diffuse, low-level staining of the tubules and mesangium,
with no concentration of immunoglobulin in the glomeruli (results not shown). Transmission electron microscopy also revealed immune complex deposition (Figure
6D), with features similar to those of lupus nephritis in
humans, including small, electron-dense deposits forming wire-loop lesions in the subendothelial basement
Lung histopathology. Pneumonitis was found in
25% of mice between ages 12 and 40 weeks, increasing
to 100% of older mice (Figure 5B), and was characterized by patches of perivascular inflammation with cellular aggregates of macrophages, lymphocytes, plasma
cells, and numerous PMN cells within alveolar walls
(Figure 6E). In severe cases, up to 50% of the normal
architecture of the lungs was obliterated, but the disease
was self-limiting, and older mice remained healthy.
Lungs of age-matched nontransgenic mice showed no
Analysis of antihistone and antinuclear antibodies
(ANAs). The histologic features suggested that the tissue
damage seen in the spontaneous autoimmune disease in
Fc␥RIIa-transgenic mice was mediated, at least in part,
by autoantibodies. Therefore, we evaluated mice for the
presence of autoantibodies known to be associated with
human autoimmune disease. Initial immunofluorescence studies with sera from most Fc␥RIIa-transgenic
mice and many older nontransgenic C57BL/6 mice
showed homogeneous nuclear staining of Chinese hamster ovary cells (results not shown). This staining pattern
was similar to that of antihistone antibody huPIA3 (29).
Sera were then tested for antihistone antibodies by
ELISA, using a mixture of purified histones. In mice
ages ⬎20 weeks, 13 of 23 had antihistone antibodies
above background levels (Figure 6F). Histone antibody
titers did not correlate with GN or arthritis incidence (r
⬍0.05). No antihistone antibodies above background
levels were seen in age-matched C57BL/6 mice or
(SJL ⫻ C57BL/6)F1 mice at ages ⬎25 weeks. None of
the mice had the other common autoantibodies, such as
anti–double-stranded or anti–single-stranded DNA or
rheumatoid factor (data not shown).
Since it was found that recombinant soluble FcR
inhibited immune complex vasculitis (3), there has been
widespread interest in the role of FcR in antibodyinduced inflammation in autoimmune diseases (5,8).
Although many studies have analyzed the role of mouse
FcR, including Fc␥RI, Fc␥RIIb, and Fc␥RIIIa, which
are common to both mice and humans, humans have a
unique FcR, Fc␥RIIa, which is absent from rodents and
was therefore not analyzed in rodent models of autoimmunity. Our data demonstrate that Fc␥RIIa-transgenic
mice are highly susceptible to passive antibody-induced
inflammation and to active (collagen-induced) and spontaneous autoimmune disease.
In CIA, there is variable susceptibility in mice,
which is linked to MHC type (24). In this study, expression of Fc␥RIIa conferred susceptibility to CIA in
strains of mice with low-susceptibility MHC (H-2b and
H-2b/s). Indeed, CIA in Fc␥RIIa-transgenic mice developed more rapidly than in the archetypal CIAsusceptible DBA/1 strain, with almost 15% of Fc␥RIIatransgenic mice developing CIA after a single dose of
collagen. These findings establish a role for Fc␥RIIa in
enhancing inflammatory responses in CIA, especially
since treatment with anti-Fc␥RIIa mAb reduced disease
severity. The immune mechanisms involved in the development of CIA have been well described, with both T
cells and anticollagen antibodies known to play major
roles (30,31).
The observation that anti-CII antibodies were
lower on day 21 after CIA induction in Fc␥RIIatransgenic mice compared with DBA/1 or nontransgenic
animals is evidence that Fc␥RIIa plays a role in increasing the sensitivity of effector cells to activation by
immune complexes. This possibility was supported by
data from the anti-CII antibody transfer model, wherein
a single dose of anti-CII antibody M2139 induced disease in 100% of the Fc␥RIIa-transgenic mice. This
contrasts with other strains, in which a mixture of 2
antibodies plus LPS was required to induce this level of
disease (20). Although the use of 1 antibody to induce
arthritis in susceptible DBA/1 mice has been reported
(32), the highest incidence was only 50% after 2 doses of
M2139 (total dose 9 mg), compared with the 100%
incidence after a single 2-mg dose in the Fc␥RIIatransgenic mice.
TNF␣ is a major clinically validated inflammatory mediator in human rheumatoid arthritis (RA) (33),
and the majority of immune complex–induced TNF␣
production from transgenic macrophages could be attributed to Fc␥RIIa activation. Endogenous mouse FcR
(Fc␥RIIIa and Fc␥RI) were responsible for the balance
of TNF␣, which was equivalent to that produced from
HAGG-stimulated nontransgenic macrophages. This
also indicates that the endogenous receptors are functionally intact and unaffected by the presence of the
transgenic receptor.
A surprising characteristic of the Fc␥RIIatransgenic mice was the spontaneous development of
disease with features of RA and systemic lupus erythematosus (SLE). Approximately 40% of older transgenic
mice had features of both RA and SLE, similar to the
“rhupus” overlap syndrome described in humans (34).
The remaining 60% of the mice had features of SLE,
e.g., endoproliferative GN, with intraglomerular accumulation of deposits (i.e., wire-loop lesions). Some
Fc␥RIIa-transgenic mice also developed mild, nonerosive inflammatory arthritis similar to that seen in SLE.
While most of the older Fc␥RIIa-transgenic mice
had antihistone antibodies, the most interesting serologic observation was elevated IgG2a levels in mice with
spontaneous severe destructive arthritis. Elevated IgG2a
was seen in other autoimmune models in mice (35,36),
and may reflect the cytokine profiles involved in leukocyte activation (37). The antibodies detected in our study
were not directed against CII and, currently, the autoantigen remains undefined.
The phenotype of the Fc␥RIIa-transgenic mice
resembles that of mice deficient in the inhibitory
Fc␥RIIb. These mice show increased susceptibility to
CIA (36), elevated levels of TNF␣ and IgG2a (38), and,
on a specific MHC background, spontaneously develop
SLE-like symptoms, including ANAs to double-stranded
DNA and DNA/histone complexes (39) and GN (40).
However, there are some fundamental differences compared with Fc␥RIIa-transgenic mice. The Fc␥RIIbdeficient mice never developed spontaneous severe destructive arthritis; they showed exaggerated antibody
responses, with elevated anti-CII antibodies in the CIA
model, whereas Fc␥RIIa-transgenic mice had low antibody levels. Also, Fc␥RIIb-knockout mice have antiDNA antibodies, but Fc␥RIIa-transgenic mice developed only antihistone antibodies. Thus, analysis of
Fc␥RIIb-deficient mice by other investigators suggests
that their phenotype was due largely to dysregulation of
B cell activation and loss of B cell tolerance (36,40)
arising from unbalanced ITAM/ITIM signaling (38).
While we cannot rule out such an imbalance, it must be
relatively subtle, since Fc␥RIIa-transgenic mice had
both activating and inhibitory receptors (Figure 1).
In contrast, Fc␥RIIb-deficient mice and cells
entirely lacked the inhibitory Fc␥RIIb, but retained a
full complement of activating receptors. Furthermore,
data from other studies (41) show that transfection of
activating FcR into cells that already express both activating and inhibitory receptors left the inhibitory
Fc␥RIIb still functional and potent. An alternative explanation for our observations is that Fc␥RIIa lowers the
threshold of immune complex activation or qualitatively
changes the response induced by pathogenic antibodies.
Indeed, evidence from human in vitro studies suggests
such a role for Fc␥RIIa in activated macrophages, where
Fc␥RI signals are partly dependent on Fc␥RIIa (42).
Nonetheless, future studies of mechanisms will be informative in this regard.
The development of destructive arthritis involves
interlinked immunologic and cytokine pathways (43,44).
It is clear that TNF␣ and IL-1␤ are major factors in
active human disease (44), and recent clinical trials have
suggested a role for B cells and possibly for immune
complexes (45,46). Although animal models have been
useful in suggesting possible mechanisms in human
disease, analysis of the role of the major and unique
activating human FcR (Fc␥RIIa) has been lacking. Our
analysis strongly suggests a role for this receptor in RA.
It is particularly interesting that Fc␥RIIa-transgenic
mice spontaneously develop destructive arthritis. This is
rare in mice, having been observed only in older males of
the susceptible DBA/1 strain (47), in strains with alterations in T cell tolerance, selection, and/or activation
(for example, K/BxN [6] and SKG [48] strains), or in
mice with engineered cytokine disturbances, such as
TNF␣-transgenic (26) and IL-1Ra–knockout (49) mice.
These studies show that expression of the unique
activating human FcR, Fc␥RIIa, is associated with spontaneous autoimmune inflammation and exaggerated reactivity to induced, antibody-dependent inflammation.
Fc␥RIIa may lower the threshold for stimulation of
effector cell function, resulting in hypersensitivity to
immune complexes in vivo, and inducing enhanced
cytokine secretion. Thus, Fc␥RIIa plays a critical role in
determining the sensitivity to autoimmunity and, as
such, provides an attractive target for the treatment of
autoimmune disease.
We thank Liliana Tatarczuch for assistance with the
transmission electron microscopy, Bill Pispalliaris for useful
discussion of the antihistone antibody analysis, Dacho Gao for
the production of the radiographs, and Dr. Peck Sze Tan
(ARI) for antibodies.
1. Dijstelbloem HM, Bijl M, Fijnheer R, Scheepers RH, Oost WW,
Jansson MD, et al. Fc␥ receptor polymorphisms in systemic lupus
erythematosus: association with disease and in vivo clearance of
immune complexes. Arthritis Rheum 2000;43:2793–800.
2. Takai T. Roles of Fc receptors in autoimmunity. Nat Rev Immunol
3. Ierino FL, Powell MS, McKenzie IF, Hogarth PM. Recombinant
soluble human Fc␥RII: production, characterization, and inhibition of the Arthus reaction. J Exp Med 1993;178:1617–28.
4. Ravetch JV, Bolland S. IgG Fc receptors [review]. Annu Rev
Immunol 2001;19:275–90.
5. De Stahl TD, Andren M, Martinsson P, Verbeek JS, Kleinau S.
Expression of Fc␥RIII is required for development of collageninduced arthritis. Eur J Immunol 2002;32:2915–22.
6. Ji H, Ohmura K, Mahmood U, Lee DM, Hofhuis FM, Boackle SA,
et al. Arthritis critically dependent on innate immune system
players. Immunity 2002;16:157–68.
7. Van Lent PL, Nabbe K, Blom AB, Holthusyen AE, Sloetjes A, van
de Putte LB, et al. Role of activatory Fc␥RI and Fc␥ III and
inhibitory Fc␥RII in inflammation and cartilage destruction during experimental antigen-induced arthritis. Am J Pathol 2001;159:
8. Hogarth PM. Fc receptors are major mediators of antibody based
inflammation in autoimmunity [review]. Curr Opin Immunol
9. Powell MS, Barton PA, Emmanouilidis D, Wines BD, Neumann
GM, Peitersz GA, et al. Biochemical analysis and crystallisation of
Fc␥RIIa, the low affinity receptor for IgG. Immunol Lett 1999;68:
10. Chuang FY, Sassaroli M, Unkeless JC. Convergence of Fc␥
receptor IIA and Fc␥ receptor IIB signalling pathways in human
neutrophils. J Immunol 2000;164:350–60.
11. Tan Sardjono C, Mottram PL, Hogarth PM. The role of Fc␥RIIa
as an inflammatory mediator in rheumatoid arthritis and systemic
lupus erythematosus [review]. Immunol Cell Biol 2003;81:374–81.
12. Maxwell KF, Powell MS, Hulett MD, Barton PA, McKenzie IF,
Garrett TP, et al. Crystal structure of the human leukocyte Fc
receptor, Fc␥RIIa. Nat Struct Biol 1999;6:437–42.
13. Powell MS, Hogarth PM. The role and use of recombinant
receptors in the investigation and control of antibody-induced
inflammation. In: van de Winkel JG, Hogarth PM, editors. Immunoglobulin receptors and their physiological and pathological roles
in immunity. Dordrecht (The Netherlands): Kluwer Academic
Publishers; 1998.
14. Hulett MD, Hogarth PM. Molecular basis of Fc receptor function
[review]. Adv Immunol 1994;57:1–127.
15. Daeron M. Fc receptor biology [review]. Annu Rev Immunol
16. McKenzie SE. Humanized mouse models of FcR clearance in
immune platelet disorders [review]. Blood Rev 2002;16:3–5.
17. McKenzie SE, Taylor SM, Malladi P, Yuhan H, Cassel DL, Chien
P, et al. The role of the human Fc receptor Fc␥RIIA in the
immune clearance of platelets: a transgenic mouse model. J Immunol 1999;162:4311–8.
18. Manger K, Repp R, Spriewald BM, Rascu A, Geiger A, Wassmuth
R, et al. Fc␥ receptor IIa polymorphism in Caucasian patients with
systemic lupus erythematosus: association with clinical symptoms.
Arthritis Rheum 1998;41:1181–9.
19. Campbell IK, Hamilton JA, Wicks IP. Collagen-induced arthritis
in C57BL/6 (H-2b) mice: new insights into an important disease
model of rheumatoid arthritis. Eur J Immunol 2000;30:1568–75.
20. Svensson L, Nandakumar KS, Johansson A, Jansson L, Holmdahl
R. IL-4-deficient mice develop less acute but more chronic relapsing collagen-induced arthritis. Eur J Immunol 2002;32:2944–53.
21. Barnes N, Gavin AL, Tan PS, Mottram P, Koentgen F, Hogarth
PM. Fc␥RI-deficient mice show multiple alterations to inflammatory and immune responses. Immunity 2002;16:379–89.
22. Tan PS, Gavin AL, Barnes N, Sears DW, Vremec D, Shortman K,
et al. Unique monoclonal antibodies define expression of Fc␥RI
on macrophages and mast cell lines and demonstrate heterogeneity among subcutaneous and other dendritic cells. J Immunol
23. Wilkins S, Gureyev T, Gao D, Pogany A, Stevenson A. Phase-
contrast imaging using polychromatic hard X-rays. Nature 1996;
Courtenay JS, Dallman MJ, Dayan AD, Martin A, Mosedale B.
Immunisation against heterologous type II collagen induces arthritis in mice. Nature 1980;283:666–8.
Matsumoto T, Ametani A, Hachimura S, Iwaya A, Taguchi Y,
Fujita K, et al. Intranasal administration of denatured type II
collagen and its fragments can delay the onset of collagen-induced
arthritis. Clin Immunol Immunopathol 1998;88:70–9.
Butler DM, Malfait AM, Mason LJ, Warden PJ, Kollias G, Maini
RN, et al. DBA/1 mice expressing the human TNF-␣ transgene
develop a severe, erosive arthritis: characterization of the cytokine
cascade and cellular composition. J Immunol 1997;159:2867–76.
Feldmann M. Development of anti-TNF therapy for rheumatoid
arthritis. Nat Rev Immunol 2002;2:364–71.
Nandakumar KS, Svensson L, Holmdahl R. Collagen type IIspecific monoclonal antibody-induced arthritis in mice: description
of the disease and the influence of age, sex, and genes. Am J
Pathol 2003;163:1827–37.
Cosgrove L. Monoclonal antibodies to platelet antigens. Melbourne (Australia): University of Melbourne; 1987.
Myers LK, Rosloniec EF, Cremer MA, Kang AH. Collageninduced arthritis, an animal model of autoimmunity [review]. Life
Sci 1997;61:1861–78.
Andersson EC, Hansen BE, Jacobsen H, Madsen LS, Andersen
CB, Engberg J, et al. Definition of MHC and T cell receptor
contacts in the HLA-DR4 restricted immunodominant epitope in
type II collagen and characterization of collagen-induced arthritis
in HLA-DR4 and human CD4 transgenic mice. Proc Natl Acad Sci
U S A 1998;95:7574–9.
Nandakumar KS, Andren M, Martinsson P, Bajtner E, Hellstrom
S, Holmdahl R, et al. Induction of arthritis by single monoclonal
IgG anti-collagen type II antibodies and enhancement of arthritis
in mice lacking inhibitory Fc␥RIIB. Eur J Immunol 2003;33:
Feldmann M, Maini RN. Anti-TNF␣ therapy of rheumatoid
arthritis: what have we learned? [review]. Annu Rev Immunol
Hahn B. Systemic lupus erythematosus and related syndromes. In:
Ruddy S, Harris E, Sledge B, editors. Kelley’s textbook of rheumatology. 6th ed. Philadelphia: W. B. Saunders; 2001. p. 1089–103.
Quattrocchi E, Dallman MJ, Feldmann M. Adenovirus-mediated
gene transfer of CTLA-4Ig fusion protein in the suppression of
experimental autoimmune arthritis. Arthritis Rheum 2000;43:
Yuasa T, Kubo S, Yoshino T, Ujike A, Matsumura K, Ono M, et
al. Deletion of Fc␥ receptor IIB renders H-2b mice susceptible to
collagen-induced arthritis. J Exp Med 1999;189:187–94.
Watson WC, Townes AS. Genetic susceptibility to murine collagen II autoimmune arthritis: proposed relationship to the IgG2
autoantibody subclass response, complement C5, major histocompatibility complex (MHC) and non-MHC loci. J Exp Med 1985;
Clynes R, Maizes JS, Guinamard R, Ono M, Takai T, Ravetch JV.
Modulation of immune complex-induced inflammation in vivo by
the coordinate expression of activation and inhibitory Fc receptors. J Exp Med 1999;189:179–85.
Bolland S, Yim YS, Tus K, Wakeland EK, Ravetch JV. Genetic
modifiers of systemic lupus erythematosus in Fc␥RIIB⫺/⫺ mice. J
Exp Med 2002;195:1167–74.
Bolland S, Ravetch JV. Spontaneous autoimmune disease in
Fc␥RIIB-deficient mice results from strain-specific epistasis. Immunity 2000;13:277–85.
Daeron M. Structural bases of Fc␥R functions [review]. Int Rev
Immunol 1997;16:1–27.
Allen JM, Seed B. Isolation and expression of functional highaffinity Fc receptor complementary DNAs. Science 1989;243:
Firestein GS. Etiology and pathogenesis of rheumatoid arthritis.
6th ed. Philadelphia: W. B. Saunders; 2001.
Feldmann M, Maini RN, Bondeson J, Taylor P, Foxwell BM,
Brennan FM. Cytokine blockade in rheumatoid arthritis [review].
Adv Exp Med Biol 2001;490:119–27.
Moore J, Ma D, Will R, Cannell P, Handel M, Milliken S. A phase
II study of rituximab in rheumatoid arthritis patients with recurrent disease following haematopoietic stem cell transplantation.
Bone Marrow Transplant 2004;34:241–7.
Edwards JC, Szczepanski L, Szechinski J, Filipowicz-Sosnowska A,
Emery P, Close DR, et al. Efficacy of B-cell-targeted therapy with
rituximab in patients with rheumatoid arthritis. N Engl J Med
Nordling C, Karlsson-Parra A, Jansson L, Holmdahl R, Klareskog
L. Characterization of a spontaneously occurring arthritis in male
DBA/1 mice. Arthritis Rheum 1992;35:717–22.
Hata H, Sakaguchi N, Yoshitomi H, Iwakura Y, Sekikawa K,
Azuma Y, et al. Distinct contribution of IL-6, TNF-␣, IL-1, and
IL-10 to T cell-mediated spontaneous autoimmune arthritis in
mice. J Clin Invest 2004;114:582–8.
Iwakura Y. Roles of IL-1 in the development of rheumatoid
arthritis: consideration from mouse models [review]. Cytokine
Growth Factor Rev 2002;13:341–55.
Без категории
Размер файла
266 Кб
development, multisystem, spontaneous, inflammation, induced, mice, iiatransgenic, autoimmune, disease, antibody, receptov, hypersensitivity
Пожаловаться на содержимое документа