close

Вход

Забыли?

вход по аккаунту

?

Spontaneous degenerative polyarthritis in male new zealand blackkn mice.

код для вставкиСкачать
171
SPONTANEOUS DEGENERATIVE POLYARTHRITIS IN
MALE NEW ZEALAND BLACK/KN MICE
KUNIE NAKAMURA, SADAO KASHIWAZAKI, KENJI TAKAGISHI, YUKIO TSUKAMOTO,
YASUO MOROHOSHI, TAKESHI NAKANO, and MASAAKI KIMURA
Histopathologic studies and radiographic analysis revealed that male New Zealand bIack/KN (NZB/KN)
mice develop degenerative polyarthritis in the joints of
the forepaw and hindpaw beginning at age 2 months.
Deposits of autoantibodies were observed on proliferating collagen fibers, nuclei of chondrocytes, and epidermal cells. Increases in the frequency of positivity for
rheumatoid factor and anti-type I1 collagen antibodies
and in the level of serum oxidation activity were noted in
these mice. The joint disease in male NZB/KN mice was
transferable to female NZB/KN mice and male BALBlc
mice by intraperitoneal injection of spleen cells from the
male NZB/KN mice. This animal model of arthritis will
be extremely useful for analyzing not only the pathogenesis of rheumatoid arthritis, but also new strategies for
its treatment, since NZB/KN mice, unlike MRLllpr
mice, do not develop severe lupus nephritis or lymphFrom the Molecular Biology Laboratory, Department of
Biochemistry, Rheumatology Group, Department of Internal Medicine, the Department of Orthopedics, and the Department of
Laboratory Animal Science, Kitasato University School of Medicine, Sagamihara, Kanagawa, and the Research Center, Taisho
Pharmaceutical Co, Ltd., Omiya, Saitama, Japan.
Kunie Nakamura, MD, DrMedSci: Molecular Biology Laboratory, Department of Biochemistry; Sadao Kashiwazaki, MD,
DrMedSci: Rheumatology Group, Department of Internal Medicine;
Kenji Takagishi, MD, DrMedSci: Department of Orthopedics;
Yukio Tsukamoto, MD, DrMedSci: Department of Orthopedics;
Yasuo Morohoshi, PhD: Department of Laboratory Animal Science; Takeshi Nakano, PhD: Department of Laboratory Animal
Science; Masaaki Kimura, PhD: Research Center, Taisho Pharmaceutical Co, Ltd.
Address reprint requests to Kunie Nakamura, MD, DrMed
Sci, Molecular Biology Laboratory, Department of Biochemistry,
Kitasato University School of Medicine, 1-15-1 Kitasato, Sagamihara, Kanagawa, 228 Japan.
Submitted for publication October 10, 1989; accepted in
revised form August 15, 1990.
Arthritis and Rheumatism, Vol. 34, No. 2 (February 1991)
adenopathy, and therefore have a longer survival period.
Autoimmune mechanisms are involved in the
development of degenerative changes of bones and
joints in patients with rheumatoid arthritis (RA). The
pathogenesis of polyarthritis has been studied extensively using various animal models. RA-like arthritis
has been induced in rats by collagen (1,2) and by
Freund’s complete adjuvant (3), in mice by proteoglycan (4),and in rabbits by proteins such as fibrin (9,
and by Escherichia coli (6). Although aging C57B1/6
mice spontaneously develop arthritis, these mice are
not suitable for studies of immunomodulated osteoarticular lesions (7) because it is uncertain whether
autoimmune mechanisms are involved in the development of the arthritis that affects these mice. Collageninduced arthritis, however, has been considered to be
a useful model for RA, since anticollagen antibodies
degrade cartilage and result in osteoarticular degeneration.
Recently, it has been found that MRLllpr mice
spontaneously develop an RA-like disease that is
associated with high titers of rheumatoid factor (RF).
However, despite clinical features that are similar to
those of human RA, it is difficult to obtain a large
amount of information about the pathogenesis of human RA from MRLllpr mice because progressive
lymphadenopathy shortens their life span (8-10). Although New Zealand black/Bl (NZB/Bl) mice are
rarely affected by bone and joint disease (ll), the
present study showed that in male NZB/KN mice,
from the age of 4 months, there is a high incidence of
spontaneous degenerative polyarthritis, which is accompanied by difficulty with walking, progressive
NAKAMURA ET AL
172
swelling and ankylosis in the paws, deformities of the
tail, alopecia o v e r t h e hip area, a n d albuminuria. In
this report, we characterize the osteoarticular lesions
in male NZB/KN mice and discuss their pathogenesis
in relation to t h e RA that occurs in humans.
Table 1. Specificity of the enzyme-linked immunosorbent assay
(ELISA) used in the determination of anti-type 11 collagen antibody
levels in murine sera*
Strain
MATERIALS AND METHODS
Animals. Male and female NZB/KN, BALB/c,
C57B1/6N, and C3H/HeN mice were obtained from the
Omura Experimental Animal Institute (Zama, Kanagawa,
Japan) and were fed water and standard mouse diet (Oriental
Yeast Co., Tokyo, Japan) ad libitum. The mice were housed
in air-conditioned facilities at the Experimental Animal
Center of Kitasato University School of Medicine. NZB/
San, the parental strain of NZB/KN, and NZB/KI mice were
obtained from Dr. A. Matsuzawa (Medical Science Institute,
University of Tokyo, Sirogane, Minato-ku, Tokyo, Japan).
Genetic analysis of NZB/KN mice. Four-month-old
male and female NZBIKN, NZB/San, and NZB/K1 mice
were genetically analyzed (by isoelectric focusing) for renal
enzymes, a hepatic enzyme, erythrocyte enzymes, major
urinary protein, serum markers, and immunogenetic markers (analysis conducted at the Experimental Animal Central
Research Institute, Takatsu-ku, Kawasaki City, Japan) (12).
Pathologic examination. Demuscled forepaws and
hindpaws, vertebrae and tails, kidneys, and other major
organs were fixed in a mixture of 3% neutralized formalin
and 1% paraformaldehyde. Decalcified forepaws, hindpaws,
tails, and vertebrae in 0.1M Tris buffer (pH 7.4) containing
10% EDTA (Wako, Osaka, Japan), as well as other soft
organs, were embedded in low-melting point (48-52°C)
paraffin (Wako) after dehydration through a graded series of
ethanol and xylene. The paraffin-embedded specimens were
stored in a refrigerator until thin sections were prepared with
a microtome. The thin sections were deparaffinized, stained
with hematoxylin and eosin, Safranin 0 (for chondrocytes),
or anti-mouse immunoglobulin subclass antibodies (antiIgM, anti-IgG1, anti-IgG3, anti-IgG2a, or anti-IgG2b), labeled with peroxidase (Tago, Burlingame, CA) or goat
anti-mouse C3 polyclonal antibody (Organon Teknika, West
Chester, PA), and visualized in a solution containing 0.1M
Tris buffer, pH 7.6, 0.05% diaminobenzidine tetrachloride
(Sigma, St. Louis, MO), and 0.001% hydrogen peroxide.
Nuclei were counterstained with methyl green.
Radiographic examination. Demuscled, fixed, whole
skeletal specimens were placed on shielded x-ray film and
exposed to low energy x-rays (SX-21; Shimadzu, Kyoto,
Japan). Radiographs were developed and were read under an
anatomic microscope (Olympus, Tokyo, Japan). Radiographs were scored according to the presence or absence of
osteolytic changes, articular abnormalities, deformities of
joint surfaces and cavities, deformities due to bone fusion,
and number of bones degraded in the tail. Mice were
grouped by age and strain, and average scores and standard
deviations were determined for each group.
Determination of anti-type I1 collagen antibody levels
in serum. Mice were killed by decapitation, and blood was
collected into Microtainers (Becton Dickinson, Rutherford,
BALBlc
C57B116N
NZB/KN
NZBlSan
MRLllpr
SJLt
Age
(months)/
sex
14lM
4lM
13lM
61M
41M
Mean f SD antibody level (A430)
Uncoated
wells
0.003
0.003
0.003
0.009
0.008
0.004
f 0.000
f 0.000
-+
0.000
t 0.001
-+ 0.000
t 0.000
Collagen-coated
wells
0.013
0.010
0.449
0.070
0.029
0.471
0.002
0.001
t 0.008
k 0.005
-+ 0.003
t 0.010
-+
2
* Sera were obtained from the strains of mice listed, diluted 1 :25 in
phosphate buffered saline (PBS) (except in the case of SJL mice),
and analyzed by ELISA. Values are the absorbance at 430 nm
wavelength.
t Anti-type I1 collagen antibodies in the SJL strain were produced
by subdermal injection of type I1 collagen in Freund’s complete
adjuvant. Antibodies were purified on a Sepharose 4B affinity
column coated with antigen and were diluted 1 :3,000 in PBS. The
initial protein concentration of the antibody was 100 pglml.
NJ). Sera were separated by centrifugation at 3,000 revolutions per minute for 15 minutes (MR-150; Tomy Seiko,
Tokyo, Japan). The sera were stored at -70°C until titration
by enzyme-linked immunosorbent assay (ELISA) (13).
Briefly, bovine type I1 collagen (100 pI dissolved in phosphate buffered saline [PBS] at a concentration of 25 pglml;
Genzyme, Los Angeles, CA) was added to a 96-well U-type
microtiter plate (Nunc, Roskilde, Denmark) and incubated at
37°C for 2 hours. After the fluid was discarded, the surface of
the plate was covered with 0.1% bovine serum albumin and
0.05% Tween 20 (100 pl in PBS) and incubated at 37°C for 30
minutes. The plates were then incubated with a serum
sample (200 pl diluted 1 : 25 in PBS) at 37°C for 2 hours. After
incubation, the wells were rinsed 5 times with PBS.
The wells were then covered with peroxidase-labeled
rabbit anti-mouse IgG antibody (Cappel, Cochranville, PA)
and incubated at 37°C for 2 hours, and the plates were rinsed
5 times with PBS. The wells were then visualized by a
reaction using 0.05M citrate-phosphate buffer, pH 5.0, containing 0.1% o-phenylenediamine and 0.001%hydrogen peroxide (Wako) at 37°C for 40 minutes. The color of the
reagents was assessed with an immunoreader (Flow, Rockville, MD). A standard curve was obtained from the pretested standard anti-type I1 collagen antibody (Collagen
Technique Training Center-COSMO Biological Co., Tokyo,
Japan), and absolute antibody titers were estimated. The
specificity of the ELISA used in the determination of antitype I1 collagen antibody titers in murine serum samples,
including the affinity-purified anti-type I1 collagen antibodies
elicited in the SJL mouse strain, was confirmed as shown in
Table 1.
Detection of RF. Sheep red blood cells (SRBC; 2 x
lo8) were rinsed in saline 3 times by centrifugation at 750g
and suspended in 0.15M phosphate buffer, pH 7.4. Ten
milliliters of SRBC was mixed with affinity-purified mouse
immunoglobulin (IgM and IgGl ;Cappel), and stirred at room
temperature for 60 minutes after the addition of 2.5% gluta-
173
POLYARTHRITIS IN MALE NZB/KN MICE
raldehyde in PBS. The SRBC were then rinsed 3 times with
PBS and suspended in 10 ml of PBS containing 0.5% normal
mouse serum from 8-week-old C57B1/6N mice. The SRBC
coated with antibody were adjusted to 1% in PBS. Serially
diluted serum samples (25 p1) were mixed with 25 pl of
SRBC coated with immunoglobulin in microtiter plates,
followed by refrigeration overnight. Levels of R F were then
estimated by titration.
Measurement of serum oxidation activity. Serum (0.1
ml) was mixed with 1 ml of o-phenylenediamine in water
(1 ml/ml; Wako) and 1.5 ml of 0.1M citrate-phosphate
buffer, pH 5.0, and was incubated at 37°C for 120 minutes.
The increase in absorbance at 430 nm (A4,,) was measured
with a spectrophotometer (type 228; Hitachi-Nissei Sangyo,
Tokyo, Japan). After the original absorbance of each reaction mixture was reduced, the increase in A,,, was calculated (14).
Injection of spleen cells into female NZBlKN mice and
male BALBlc mice. Spleen cells (1 X 10') were isolated from
4-month-old male or female NZB/KN mice and were injected intraperitoneally into female NZBiKN mice and male
BALBlc mice at 2-month intervals. In addition, an aliquot of
spleen cell suspension in Hanks' balanced salt solution
(HBSS) was incubated at 37°C for 30 minutes with antimouse Thy-1.2 antibody (Cedarlane, Hornby, Ontario, Canada) at a dilution of 1 : 160, with a 1/30 volume of guinea pig
serum as the complement source. Cells were rinsed 3 times
with HBSS, and were then injected into recipient mice.
Recipient mice were observed for symptoms of arthritis in
forepaw and hindpaw joints and for development of tail
deformity. Mice showing clinical signs of arthritis underwent
pathologic examination, which included enzyme-labeled
anti-immunoglobulin staining of tissue specimens.
Table 2. Genetic analysis of NZB mice*
~~~
~
Chromosome,
locus
Genetic marker
NZB/KN mice
NZB/San mice
NZB/K1 mice
1
Idh-1
Pep-3
Akp- 1
2, Hc
3, Car-2
4
Mup-1
Gpd- 1
5 , Pgm-1
6, Ldr-1
7
Gpi-1
Hbb
8
Es-1
Es-2
9
Thy- 1
Mod- 1
Trf
11, Es-3
17
H-2K
H-2D
a
a
a
C
C
C
b
0
a
b
0
a
b
a
b
b
a
a
b
b
bt
b
b
a
a
d
a
d
b
b
b
b
b
b
b
b
b
b
b
b
b
b
C
C
C
d
d
d
d
d
d
a
It
a
a
st
at
RESULTS
* Idh-1, Pep-3, Gpd-1, Pgm-1, Es-1, Es-2, Mod-1, and Es-3 are
marker enzymes in the kidneys. Akp-1 is a marker enzyme in the
liver. Hc and Trf are marker enzymes in serum. Car-2, Ldr-1, and
Gpi-1 are marker enzymes in erythrocytes. Mup-1 is a major urinary
protein marker. Hbb is a hemoglobin beta chain marker. Thy-1,
H-2K, and H-2D are immunogenetic markers. The genetic markers
in NZBiKN mice were the same as those in the NZBlBl standard
strain.
t Indicates a genotypic change in this strain.
Genetic features. The NZB/KN strain is a subline of NZB/San, which had been bred at the Medical
Science Institute, Tokyo University, and donated 12
years ago to the Kitasato University School of Medicine. Genetic monitoring of NZB/KN mice clearly
showed the absence of contamination or genetic variation. Each genetic marker was isologous with that in
the standard NZB strain developed by Bielshofsky in
1956 (Table 2). NZB/San, the parental strain of NZBI
KN, showed the same markers as those in NZB/KN.
Another strain, NZB/Kl, demonstrated minor changes
in marker proteins; these mice had lymphomas but had
no osteoarticular lesions.
Radiographic observations. Abnormal radiographic changes first became evident at the tips of the
phalanges of both forepaws and hindpaws at the age of
2 months. Degeneration of bones and joints progressed
with time, and ankylosis of the hindlimb joints began
at 6 months of age (Figure 1). As shown by the
radiographic scores (Figure 2), progressive degenera-
tive changes of bone were noted in male NZB/KN
mice. Up to the age of 5 months, MRLllpr mice
showed the same radiographic changes seen in male
NZB/KN mice (Figure l), but the progressive growth
of lymphomas in the MRLllpr mice prevented further
observation. In male NZB/San mice and in female
NZB/KN mice, comparable radiographic changes
were not observed, even up to the age of 17 months.
Pathologic findings. At 2 months of age, the
following features were observed on hematoxylin and
eosin-stained sections of footpads from these
NZBlKN mice:hypertrophy and hyperplasia of synovial lining cells, infiltration of lymphocytes and
mononuclear cells into connective tissue beneath the
synovium, osteolytic lesions and fibroblastic invasion
of bone, erosion of the cartilage layer, irregularity of
joint surfaces, and abnormal proliferation of chondrocytes in extraarticular areas (Figure 3). These changes
became more severe by 6 months of age, when osteo-
NAKAMURA ET AL
174
Figure 1. Radiographs of the hind limbs and caudal vertebrae of male and female NZB/KN, NZB/San, and
MRLllpr mice. Abnormalities are indicated by arrows. 1.1, Two-and-a-half-month-old male NZB/KN mouse.
1.2, Four-month-old male NZBiKN mouse. 1.3, Six-month-old male NZBiKN mouse. 1.4, Fourteen-monthold male NZB/KN mouse. 1.5, Four-month-old male MRL/lpr+ mouse. 1.6, Six-month-old male NZB/San
mouse. 1.7, Six-month-old female NZB/KN mouse. 1.8, Fourteen-month-old female NZB/KN mouse.
lysis, osteophytes, and bone fusion, calcification, pannus formation, progressive erosion of the cartilage
layer at the joint surfaces, abnormal extraarticular
proliferation of chondrocytes, further thickening and
degeneration of synovial lining cells, and infiltration of
mononuclear cells and fibroblasts into the degenerative lesions associated with ankylosis of the joints of
the forepaw and hindpaw were observed (Figure 3).
175
POLYARTHRITIS IN MALE NZB/KN MICE
Sex
(m)
C57BL/6N
M
8
Balb/c
M
8
F
4
M
3
M
5
M
8
S t r ai n
NZB/San
50
100
150
NZB/KN
NZB/KN
Figure 2. Radiographic scores (mean 2 SD) in groups of mice of various strains. The osteoarticular changes
assessed are described in Materials and Methods. One point indicates 1 change.
Skin erosions were occasionally evident on the hips
and tail, and were associated with inflammatory infiltration and spongy degeneration of the bones of the tail
(Figure 1). These morphologic changes progressed,
and by the age of 14 months, osteonecrosis, calcification, and reconstitution of osteoarticular conformations could be observed (Figure 3).
Extraarticular proliferation of chondrocytes was
confirmed by Safranin 0 staining, which showed massive proliferation of chondrocytes in areas where
osteophyte formation and bone fusion had progressed
(Figure 4). Serial sections were stained by an immu-
nohistochemical method using anti-S100 protein antibody. Chondrocytes showed positive staining with this
method as well (data not shown).
Immunohistochemical findings. Paraffin-embedded thin sections of footpads were stained with
anti-mouse IgGl labeled with peroxidase. Nuclei of
chondrocytes, surfaces of collagen bundles, striated
skeletal muscles, proliferating collagen fibers in degraded connective tissue, hair root cells, and epidermal cells showed positive staining with this antibody
(Figure 4). The staining became more distinct with
age. Similar results were obtained with anti-mouse C3
176
NAKAMURA ET AL
did not show the positive staining observed in the male
NZB/KN mice.
RF titers. SRBC coated with affinity-purified
mouse IgM and IgG were used to detect RF. RF
positivity was found in each NZB strain, including
female NZB/KN mice without articular disorders
(mean titer 1 : 128). BALB/c mice had very high RF
titers at the age of 17 months (mean titer 1 :64). We
therefore concluded that the RF titer cannot be used as
a definitive marker for the development of autoimmune arthritis.
Figure 3. Histopathologic analysis of footpad sections from male
NZB/KN mice. 3.1, Hypertrophy and hyperplasia of synovial lining
cells from the tarsi of the footpad in a 3-month-old mouse. 3.2,
Invasion of fibroblasts into the phalanges of the footpad in a
2-month-old mouse. 3.3, Abnormal proliferation of chondrocytes in
a joint cavity in a 2-month-old mouse. 3.4, Osteonecrosis of the tip
of a phalanx and proliferation of fibroblasts in a 2-month-old mouse.
3.5, Bone fusion and abnormal extraarticular proliferation of chondrocytes in a 6-month-old mouse. 3.6, Infiltration of mononuclear
cells around muscle bundles in the footpad of a 6-month-old mouse.
3.7, Erosion, destruction of thejoint surface, and deposition of fibrin
in the joint cavity in a 6-month-old mouse. 3.8, Osteonecrosis,
calcification, and destruction of joint conformation in a 14-monthold mouse. (Hematoxylin and eosin stained, original magnification
x 100.)
antibody staining. Tissue from 6-month-old female
NZB/KN, male NZB/San, male NZB/Kl, male
BALB/c, male C3H/NeN, and male C57BL/6N mice
Figure 4. Immunohistochemical and histologic analysis of footpads
of 6-month-old NZB/KN mice, stained with anti-mouse IgGl antibody (4.1-4.5) or with Safranin 0 (4.6). 4.1, Specific staining of
nuclei of chondrocytes in the heel bone and Achilles tendon (original
magnification x 200). 4.2, Striated skeletal muscles and surfaces of
collagen bundles (tendon), showing positive staining (original magnification X 200). 4.3, Collagen fiber proliferation in degraded
connective tissue of the footpad, showing strong staining (original
magnification x 100). 4.4, Epidermal layers of the tip of a phalanx,
showing weak but positive staining (original magnification x 100).
4.5, Hair root cells and epidermal layers, showing positive staining
(original magnification x 200). 4.6, Bone fusion, abnormal extraarticular proliferation of chondrocytes, and proliferation of synovial
lining cells, evidenced by positive staining (original magnification x
100)*
POLYARTHRITIS IN MALE NZB/KN MICE
Table 3. Anti-type I1 collagen IgG antibodies in murine sera
determined by enzyme-linked immunosorbent assay
Strain, age
(months)/sex
No. positive1
no. tested
Antibody
level
(I*ddl)*
014
1I4
7112
013
1I5
017
113
013
015
215
116
013
18.0
41.1 f 21.0
39.0
21.0
15.6 2 19.1
462.0
-
414
014
32.0
212
011
212
I12
215
63.0
63.0 f 0.0
47.0
18.8 f 24.4
112
415
213
41.O
36.7 f 13.3
32.5 f 8.5
0110
-
0110
-
0110
-
NZBIKN
1.5lM
2.0lM
4.0lM
6.0lM
7.51M
14.0lM
16.0lM
3.5lF
4.0lF
6.0lF
14.0lF
17.0lF
MRLllpr
4. OIM
4.0lF
NZBISan
3.5lM
5.0lM
5.5lM
7.51M
9.0lM
NZBiKl
5.01M
5.51M
7.51M
C3HlHeN
4.0lM
C57B116N
4.0lM
BALBlc
14.0lM
~~
?
5.2
-
5
-
9.0
~~~
* Where antibodies were detected in more than 1 animal, values are
the mean k SD.
Levels of anti-type I1 collagen antibody. At the
age of 3 months, male NZB/KN mice produced serum
anticollagen autoantibody at a mean level of -18
pg/dl. The amount of autoantibody increased to a
mean of 78 pgldl by the age of 5 months, but 5 of 12
sera from male NZB/KN mice were negative by
ELISA. Nonarthritogenic mouse strains, such as female NZB/KN, male NZB/San, and NZB/KI, also
produced these antibodies. However, young male
C3H/HeN, young male C57B1/6N, and old male
BALB/c mice did not produce these antibodies (Table
3). It is evident from these results that production of
anti-type I1 collagen autoantibodies cannot serve as a
definitive marker for the development of autoimmune
arthritis in these mice.
Serum oxidation activity. Five-month-old male
NZB/KN mice showed significantly higher levels of
serum oxidation activity than did male NZB/San,
female NZB/KN, male C3H/HeN, or male C57B1/6N
177
mice (data not shown). However, male NZBiKl mice
showed higher levels than male NZB/KN mice, suggesting that this activity is also not a useful marker for
the development of joint disease.
Transfer of arthritis. Four to 6 months after the
injection of spleen cells of male NZB/KN mice into
female NZB/KN mice, the female mice showed ankylosis of the joints of the feet and tail deformities.
Pathologic examination revealed the same histologic
changes that were seen in male NZB/KN mice with
spontaneous arthritis. These changes were observed in
all 10 of the female NZB/KN mice injected with spleen
cells from male NZB/KN mice, and in none of the 10
female NZB/KN mice injected with spleen cells from
other female NZB/KN mice. However, female
NZB/KN mice died of autoimmune diseases other
than RA; thus, they could be used in these experiments only for a period of 8 months.
Experiments were also performed using male
BALB/c mice as the recipients; the H-2 haplotype in
these mice was isologous to that in NZB/KN mice.
Ten of 14 male BALBic recipients showed swelling
and ankylosis of the foot joints and tail deformities 10
months after a regimen of 4 intraperitoneal injections
of spleen cells from male NZB/KN mice. Pathologic
examination demonstrated histologic features similar
to those of 7-month-old male NZB/KN mice. Osteoarticular disease did not develop during the 12-month
observation period in any of 6 recipient mice injected
with spleen cells that had been treated with anti-mouse
Thy-1.2 antibody and complement. These results indicate that T cells play a crucial role in the development
of bone and joint disease in male NZB/KN mice.
DISCUSSION
The NZB strain of mice rarely manifests osteoarticular disease (11), although it has been used as
an animal model for autoimmune diseases such as
systemic lupus erythematosus and autoimmune hemolytic anemia. In this study, we demonstrated that
males of the mutant strain NZB/KN provide a highly
useful animal model for analyzing the pathogenesis of
joint disease, based on the following characteristics: 1)
sex association, 2) development of clinically evident
degenerative lesions of the forepaws and hindpaws, 3 )
longevity (24 months) despite the presence of osteoarticular disease, and 4) development of joint disease the
progression of which can be followed radiographically.
NAKAMURA ET AL
178
Table 4. Comparison of clinical symptoms and pathologic findings in murine arthritis and human
rheumatoid arthritis (RA)*
MRLllpr
mice
Acute recurrent arthritis
Peripheral
Vertebral
Chronic degenerative arthritis
Progressive destructive
polyarthritis
Dermatitis
Genital lesions
Ophthalmologic disease
Cutaneous or subcutaneous
nodules
Rheumatoid nodules
Perivasculitis
Lymphoma
Acute or subacute synovitis
Mononuclear cell infiltration
Pannus invasion into joint
cavities and bones
Tendinitis
Ankylotic changes
Fibrinous
Bone
*
- = almost never occurs;
NZB/KN
male mice
Rat adjuvant
arthritis
Human
RA
-
-
++
+
++
++
+
++
+
++
-
+-
++
+
++
++
++
-
-
-
-
++
-
++
++
++
-
++
++
++
++
++
+-
MRL/n
mice
+++
++
++
++
++
+
++
+
++
+
+
-
++
++
-
+
-
++
++
++
++
+
+
+ = rarely occurs; ++ = commonly occurs.
Table 4 shows some of the symptoms and
pathologic findings in NZB/KN mice compared with
those in other strains of mice and in patients with RA
(15). Features in male NZB/KN mice are similar to
those in humans with RA in the following respects: 1)
swelling of joints, 2) polyarthritis, 3) symmetric and
bilateral joint lesions, 4) decalcification seen in hematoxylin and eosin-stained specimens, 5 ) presence of
RF, 6) hyperplasia and hypertrophy of synovial lining
cells, 7) infiltration by lymphocytes and plasma cells,
8) overproduction of fibrin in joint cavities, and 9)
necrosis in bone and cartilaginous tissue. All of these
features are consistent with the criteria for RA in
humans (16,17). However, despite the presence of
these typical clinical symptoms in male NZB/KN
mice, there was no laboratory parameter that was
found to be a definitive marker for the development of
autoimmune arthritis in these mice. Anti-type I1 collagen antibodies, serum oxidation activity, and RF, for
example, were not specific to NZB/KN mice, but were
also found in other, nonarthritic NZB strains. However, the case is similar in humans, and diagnosis of
RA is thus made on the basis of a set of criteria (16,17).
Therefore, the fact that no definite parameter could be
found for diagnosing joint disease in male NZB/KN
mice does not mean that this strain is unsuitable as a
model for the study of human RA.
Deposits of autoantibodies in various tissues
were observed using immunohistochemical techniques. Autoantibodies seen on chondrocytes, striated
skeletal muscle, and collagen fibers may be considered
as evidence of autoimmune osteoarticular disease,
since they were closely correlated with the development of bone and joint deformity. However, such a
correlation was not found with autoantibodies to hair
root and epidermal cells. Since multiple autoantibodies are found in NZB mice, these autoantibodies may
be involved in the development of complex autoimmune phenomena in this strain, and possible also in
the pathogenesis of osteoarticular disease. Anti-red
blood cell antibodies, anti-DNA antibodies, deposits
of immune complexes on the basement membrane of
glomeruli, and antilymphocyte antibodies could be
shown to have an important role in the development of
osteoarticular disease, but this is unlikely based on
evidence gathered to date. We have conducted other
studies concerning immunologic abnormalities in
NZB/KN mice (18,19), and the results may contribute
to an elucidation of the etiopathogenesis of RA.
ACKNOWLEDGMENTS
The authors thank Keiko Nomoto (Department of
Biochemistry, Kitasato University School of Medicine) for
POLYARTHRITIS IN MALE NZB/KN MICE
assistance in conducting the experiments and K. Nakayama
(Experimental Animal Center, Kitasato University School of
Medicine) for help in preparing the radiographs. We also
thank Dr. A. Matsuzawa (Medical Science Institute, University of Tokyo) for preparing the NZB strains. We gratefully
acknowledge Dr. Y. Hosoda (Department of Pathology,
Keio University School of' Medicine) and Dr. S. Ikehara,
(First Department of Pathology, Kansai Medical College) for
their valuable comments on the pathologic findings.
REFERENCES
1. Trentham DE, Townes AS, Kang AH: Autoimmunity to
type I1 collagen: an experimental model of arthritis. J
Exp Med 146:857-868, 1977
2. Cremer MA, Townes AS, Kang AH: Collagen induced
arthritis in rodents: a review of clinical, histological and
immunological features. Ryumachi 24:45-56, 1984
3. Pearson CM, Wood FD: Studies of arthritis and other
lesions induced in rats by the injection of mycobacterial
adjuvant. VII. Pathologic details of the arthritis and
spondylitis. Am J Pathol 42:73-80, 1963
4. Glant TT, Mikecz K, Arzoumanian A, Poole AR: Proteoglycan-induced arthritis in BALB/c mice: clinical
features and histopathology. Arthritis Rheum 30:201212, 1987
5. Dumonde DC, Glynn LE: The production of arthritis in
rabbits by an immunological reaction to fibrin. Br J Exp
Pathol 43:373-380, 1962
6 . Abruzzo JL, Christian CL: The induction of rheumatoid
factor-like substance in rabbits. J Exp Med 114:791-812,
1961
7. Silberberg M, Silberberg R: Age changes of bones and
joints in various strains of mice. Am J Anat 68:69-95,
1941
8. Murphy ED, Roths JB: Autoimmunity and lymphoproliferation: induction by mutant gene lpr and acceleration
by a male association factor in strain BXSB mice,
Genetic Control of Autoimmune Diseases. Edited by N
Rose, P Pigazzi, N Warner. New York, Elsevier NorthHolland, 1978
9. Hang I,, Theofilopoulos AN, Dixon FJ: A spontaneous
rheumatoid arthritis-like disease in MRL/1 mice. J Exp
Med 155:169&1701, 1982
179
10. Rordorf C, Schneibli HP, Baltz ML, Tennent GA, Pepys
MB: The acute phase response in (NZB x NZW)F, and
MRL/I mice: contrasting patterns resembling those in
human systemic lupus erythematosus and rheumatoid
arthritis, respectively. J Exp Med 156:1268-1273, 1982
11. Wigley RD, Couchman KG, Maule R, Reay BR: Degenerative arthritis in mice: study of age and sex frequency
in various strains with a genetic study of NZB/BI,
NZYiBl, and hybrid mice. Ann Rheum Dis 36:249-253,
1977
12. Kato H: Immunogenetic monitoring system in the
mouse, Immunogenetics of the Mouse: Perspective and
Analytical Technique (abstract in English). Edited by W
Moriwaki, DW Bailey. Japan, Soft Science Publications,
1988
13. Takagishi K, Kaibara N, Hotokebuchi T, Arita C, Morinaga M, Arai K: Effect of cyclosporin on collagen
induced arthritis in mice. Ann Rheum Dis 45:339-344,
1986
14. Nakamura K, Endo H, Kashiwazaki S: Serum oxidation
activities and rheumatoid arthritis. Int J Tissue React
IXt307-316, 1987
15. Pearson CM: Arthritis in animals, Arthritis and Allied
Conditions. Edited by DJ McCarty. Philadelphia, Lea &
Febiger, 1979
16. Ropes MW, Bennett GA, Cobb S, Jacox R, Jessar RA:
1958 revision of diagnostic criteria for rheumatoid arthritis. Bull Rheum Dis 9: 175-176, 1958
17. Blumberg BS, Bunim JJ, Calkins E, Pirani CL, Zvaifler
NJ: ARA nomenclature and classification of arthritis and
rheumatism (tentative). Arthritis Rheum 7:93-97, 1964
18. Nakamura K, Yoshii A, Akahoshi T, Kashiwazaki S,
Kawakami M: Regulation of macrophage phagocytosis
of syngeneic erythrocytes by T-cell subsets from NZB
mice: differential effects of T-cells from young and old
mice. Immunology 46561-573, 1982
19. Nakamura K, Akahoshi T, Yoshii A, Kashiwazaki S:
Generation of anti NZB red blood cell antibody-forming
plasma cells from bone marrow cultures of syngeneic
and allogeneic mice: functional modulation of helper
T-cell subsets in autosensitization. Immunology 48:57&
586, 1983
Документ
Категория
Без категории
Просмотров
3
Размер файла
1 966 Кб
Теги
spontaneous, degeneration, blackkn, mice, malen, polyarthritis, new, zealand
1/--страниц
Пожаловаться на содержимое документа