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Nineteenth rheumatism review. Part-III

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ing was not found while social, dietary and
racial factors appeared (D21). On the other hand, it was concluded from a study of
institutionalized mental patients who had
an increased incidence of osteoporosis that
physical activity may be important in senile
osteoporosis. Again the greater frequency
of the disorder in white women as compared to Negro women or men of either
race was confirmed (E35). An unusual
form of osteoporosis in Johannesburg Bantu males, associated with severe iron overload and scurvy, has been studied further
(L94). Japanese women with hypertension, similarly have an increased prevalence of osteoporosis with age (K9). Comparison of roentgenographic estimates of
bone density of vertebrae from women residing in areas of North Dakota where the
fluoride content of drinking water is either
low (0.15-0.30 ppm) or high (4-6 ppm)
has indicated about one half the frequency
of osteoporosis for the high fluoride waterdrinkers (H48).
In 9 of 13 patients studied with osteoporosis, intestinal lactase deficiency was detected. It was suggested that in such patients osteoporosis may have developed, at
least in part, through voluntary, long-term
dietary restriction of milk ingestion to
avoid symptoms of lactose intolerance
Experimentally, disuse osteoporosis can
be studied in laboratory animals. In the
cat, fluoride ingestion did not influence the
development of the disorder leading to the
suggestion that the reported beneficial
effects of fluoride therapy may be the result
of increased bone formation rather than of
reduced bone resorption (M113). In the
dog, mineral is lost in direct proportion to
collagen and the major portion of lost bone
tissue is replaced by water, fat and noncollagenous protein (K84).
I n a study of 135 women over 50 years
old who were undergoing periodic health
survey or complained of varying degrees of
chronic back pain, a direct correlation between roentgenographically assessed bone
density and bone marrow mast cell counts
has been established. It was theorized that
mast cells, which contain heparin, may
have a role in the pathogenesis of osteoporosis and other demineralizing disorders
of bone (F38). [Of interest are the observations that a variety of anionic polyelectrolytes, including heparin, may induce osteopor.osis in humans and experimental
animals. It is far from clear, however, how
these compounds mediate bone matrix
t u r n e r in view of their demonstrated effects on collagen aggregation, collagenase
activity and intermediary metabolism. Ed]
Utilizing the technic of quantitative microradiography as a measure of resorption and
formation of bone from a group of 26
patients with idiopathic osteoporosis, a significant positive correlation was found between bone resorption and serum phosphate values. No such relationship was
found between serum calcium or alkaline
phosphatase values (K50).
Idiopathic juvenile osteoporosis in children is an apparently unusual form of
osteoporosis without known cause. An additional case report and review emphasized
that the majority of such patients undergo
spontaneous remission, so that active therapy is not indicated. Quantitative microradiography of bone from the iliac crest, prior
to remission, showed normal bone formation and an almost threefold increase in
the level of bone resorption (C77).
A simple radiographic method for the
quantitation of osteoporosis has been
presented and applied to the osteoporosis
of RA as influenced by age, sex and ,therapy with corticosteroids. Cortical thickness
of the radius and spine density were measured. While the method was suitable for
Arthritis and Rheumatism, Vol. 13, No. 5 (September-October 1970)
assessing differences in bone density between groups of individuals, it was unsuitable for examining paired measurements in
individuals radiographed at different times
(S29). In patients with RA treated with
corticosteroids, the incidencx of osteoporosis was increased in both sexes, mainly in
patients over the age of 50; however, spinal
compression fractures were no greater
(S31). During an observation period of 4
years, 33 patients with RA were determined to have a significant correlation between the progress of osteoporosis of the
proximal phalanges, assessed radiographically, and grip strength, destructive
changes and sedimentation rate (V20).
A new bone density index for the diagnosis of senile osteoporosis was described
and applied to a group of control and test
subjects. It is based upon a careful radiographic assessment of calcium content of
the cortex of the fourth metacarpal of the
nondominant hand. An index figure reflecting high density and narrow cortex was
found; this is in agreement with the previously described calcification of the Haversian canal system in senile osteoporosis
(P77).I n an attempt to measure the effects
of therapy (calcium supplements and/or
anabolic compounds) on osteoporosis of
the spine various criteria were reassessed,
namely (a) the photographic density of
vertebral bodies relative to that of adjacent
soft tissues, (b) relative prominence of vertical trabeculae, (c) thickness of the anterior cortex of the vertebrae, (d) endplate
thickness and photographic density, and
(e) vertebral biconcavity. It was concluded
that none of these methods proved reliable
ence of fluoride, but bone formation or
resorption rates were unchanged, judging
by 45Ca turnover studies (L90). On the
other hand, the effects of therapy with
sodium fluoride on 4 patients with osteoporosis were assessed by microradiography
of biopsy specimens of bone tissue; it was
concluded that bone mass was increased
due to stimulated bone formation in the
presence of reduced resorption (532, J33).
[With the less than convincing information which has appeared about fluoride as
a therapy for idiopathic osteoporosis combined with its potential toxicity, much
more investigation is required before it can
be regarded as safe or eficacious for general use. Ed]
A distinctive clinical syndrome is emerging and is supported by additional reports
of adult patients with pain and roentgenologic evidence of transient osteoporosis of
the hip. Recovery has occurred within a
matter of months and therapy is supportive
Legg-Perthes Disease. Idiopathic avascular necrosis of the femoral head, occurring
in childhood, has been termed LeggPerthes disease. The etiology is unknown.
The attack rate is approximately one in
750 male children and one in 400 female
children. The disease has two striking characteristics, clinical onset within a relatively
narrow age range, usually between 4 and 8
years, and predominance in boys (h1129).
It is not now known to what extent factors
associated with avascular necrosis of known
etiology (trauma, infection, vascular dis(D35).
Conflicting reports regarding the effects turbances, etc) are involved. In one series of
of fluoride therapy on calcium retention in 168 cases (M129) the only statistically sigosteoporosis continue to appear (B68). nificant finding was the excess of children
The total exchangeable calcium was shown of low birth weight among those with
to be significantly reduced under the influ- Legg-Perthes disease. The data indicated
Arthritis and Rheumatism, Vol. 13, No. 5 (September-October 1970)
that male infants of birth weight under 51/,
pounds were five times as likely to develop
disease as male infants weighting 8y2
pounds or more at birth. Treatment of this
condition is usually conservative-some degree of immobilization (K28). T h e abduction-brace support with nonweightbearing
appeared to offer the best advantage for
eventual improvement and recovery.
The mechanisms involved in LeggPerthes disease were investigated (KSO).
Contrary to a prior report, it was concluded that an increase in anteversion of
the hip was not an adequate explanation
for the disease. Arthrography of the hip
was not recommended in the routine workup of patients with this condition (K29),
although in the later stages such visualization could demonstrate fragmentation and
reparative stages as well as flattening of the
femoral head and poor congruity between
the head and the socket, with altered acetabular cartilage thickening. T h e value of
this procedure was limited.
In another report, trochanteric osteochondritis (16 cases) was reported and
discussed (T40). The clinical features were
pain and tenderness over the greater trochanter. The clinical course was essentially
benign and the condition was self-limiting
with an excellent prognosis.
Relapsing Polychondritis. Relapsing polychondritis is a disease of which the main
features are episodic inflammation, followed by dissolution of cartilage throughout the body. There is frequently inflammation of the iris and sclera. Approximately 50 cases have been described in the
world’s literature since the first reported
case in 1923. Recent reports describe 3
additional cases (H64, J22, T47). A
51-year-old woman had typical involvement
of the ears, nose and the larynx and seemed
to respond reasonably well to steroid therapy. The first case reported from Denmark
described (522) a 45-year-old woman who
died of respiratory failure after collapse
of the upper respiratory passages. Histochemical staining indicated a loss of acid
mucopolysaccharides from the cartilage
matrix. As in previous studies, the authors of this paper could not conclude
whether this loss of matrix material was the
primary defect or was secondary to antecedent perichondritis. Additional studies
will be required to define the pathogenetic
mechanisms involved in this widespread
dissolutive disease of cartilage.
Knowledge of the chemical composition
and fine structure of amyloid depends on its
quantitative extraction from human tissues
and its purification from contaminants. Recent reports discussed methods, analyses,
and criteria for purity (A47, K65, P74). I n
one method, nonamyloid protein and salts
were first removed, then the amyloid was
homogenized in water to produce a clear,
water-soluble preparation (P75). Purified
preparations showed positive staining with
Congo red and had similar sedimentation
coefficients; however, analyses in different
laboratories revealed variations in carbohydrate content. Indeed, the nature of the
association of carbohydrate with amyloid
remains unresolved. It has generally been
felt that glycosaminoglycans (acid mucopolysaccharides) do not constitute a significant part of purified amyloid, yet a recent
report noted the presence of a uronic acidcontaining component in purified amyloid
fibrils (P37). Amyloid-laden tissues themselves seem to show a general increase in
total glycosaminoglycan content, due primarily to heparitin sulfate (D3). Organs
from mice receiving casein to produce
amyloidosis showed an increased content of
heparitin sulfate (D4). I n amyloid-laden
Mhritia and Rheumatism, Vol. 13, No. 5 (Saptember-October 1970)
human tissues, lipid was also evident, as
revealed by oil red 0 stains, but lipid
content of fibrils fell progressively, with
purification to less than 3%, a quantity
regarded as probable contamination
Casein continues to be perhaps the most
widely used agent to induce amyloidosis in
animals. When combined with complete
Freund’s adjuvant, two spaced intraperitoneal injections sufficed to producle
amyloid in C,H mice (S151). A diet high
in casein also favored amyloid deposits
(W56). Pretreatment with cortisone s u p
pressed amyloid formation in mice injected
with salmonella in oil (L22). E coli endotoxin was shown to be capable of inducing amyloidosis (B34). [Unpublished data
suggest that the amyloid inducing potential
of casein may be due to trace contamination with endotoxin-!ike materials. Ed] Intrabronchial application of 20-methylcholanthrene to rabbits produced renal amyloidosis with nephrosis and amyloid deposits in other organs as well (H69). Fragments of spleen from mice sensitized by
two injections of bacteria in oil, but in
general without amyloid deposits, were
maintained in culture for several weeks
and in some cases showed positive staining with thioflavine or Congo red. This
suggested to the authors intracellular formation of amyloid (L21). [Serial examination of explants over several weeks
should be made to provide more quantitative information to support this conclusion.
Ed] When splenic tissue from mice, sensitized with an amount of casein insufficient
to produce amyloid, was transferred to mice
treated with x-ray or nitrogen mustard,
suboptimal injections of casein into the
recipients caused widespread amyloidosis
(H33, H101). Human amyloid deposits
were said to consist of two morphologic
components that were separated on the
basis of differences in sedimentation in the
ultracentrifuge, and differences in solubility in alkaline buffers. The periodic rod,
composed of stacked pentagonal unit structures or “doughnuts,” showed a noncrystalline x-ray diffraction pattern and was a
globular glycoprotein, deficient in tryptophan and methionine. The fibril was composed of laterally aggregated unit structures of filaments with a “pleated sheet”
structure on x-ray diffraction. Peptide maps
of the two components indicated no detectable fragments in common, hence these
components were thought to be separate
and distinct (El, G27). A numbcr of workers described the fine structural features of
amyloid fibrils in cells of the spleen, liver
and bone marrow, and emphasized the role
of reticuloendothelial cells in amyloid
formation (B57, R5, Ul). In the human
kidney, the mesangial area appeared to be
the primary site associated with the deposition of amyloid fibers (S85). However, in
the guinea pig injected with casein, or in
the hamster infected with Leishmania,
amyloid deposits were extensive in the peritubular interstitium, but minimal i n the
glomerulus (S83, S86). Distinctions were
made between the amyloid deposits in
familial Mediterranean Fever (FMF) or
protracted suppuration, with those in multiple myeloma; in the former, rectal biopsies revealed amyloid filaments packed in
the intima of arterioles (so-called perireticulin amyloidosis); in the latter, filaments
were packed within collagen bundles of the
submucosa (so-called pericollagen amyloidosis) (S135).
There now seems to be some agreement
that the amyloid fibril itself, the major
component of amyloid, is probably nonimmunogenic or very weakly so (S150), while
the periodic rod, a relatively minor component whose relationship to amyloid itself
is not clear, is antigenic. The periodic rod,
M h r b a d Rheumatism, Vol. 13, No. 5 (September-October 1970)
swine (R37). This strain of mycoplasma
appeared to produce polyarthritis but no
serosal inflammation, as occurs with other
mycoplasmal strains. T h e organism a p
peared to be of low pathogenicity and/or
inferior antigenicity which may have accounted, in part, for the relatively mild
polyarthritis that was seen in the infected
animals. I n a companion study (R36), the
mycoplasmal organisms were demonstrated
by the indirect immunofluorescent antibody technic to be present in the synovial
and adjacent tissues, even in the relative
absence of microscopic lesions in the joints
of some animals. It was furthermore noted
that in young pigs the mycoplasma could
be isolated from joint fluid of clinically and
microscopically normal joints. However in
older pigs (45 kg and over), as the organism invaded the synovial tissues, microscopic and gross lesions appeared as well as
evidence of the presence of the organisms
in those tissues. The clinical and pathologic features of two types of porcine polyserositis and polyarthritis were described and
compared in another report (K73). Glasser's disease due to infection with Haemophilus suis often produced a very malignant infection with high fever and a short
course as well as either complete recovery
or death with polyserositis, polyarthritis
and usually purulent meningitis. The secExperimental Models of Arthritis and ond condition, mycoplasmosis of pigs,
Related Connective Tissue Diseases
demonstrated mild symptoms, a low grade
and Some Naturally Occurring
fever, a low morbidity and mortality and a
Counterparts in Animals
prolonged course, oftentimes with retardaINFECTIOUS ARTHRITIS IN ANIMALS
tion of growth. Pathologically there was
T h e majority of publications on this polyserositis and low grade polyarthritis
topic, which appeared in the literature but rarely meningitis. T h e appropriate orduring the past 2 or 3 years, were related to ganisms could be isolated from the joints
the role of mycoplasma in the production and certain other tissues and body fluids in
of an infectious polyarthritis in various these two conditions. It was again emspecies of animals. A strain of mycoplasma, phasized that mycoplasma, M gallisepticum
M hyoarthrinosa, has been shown to be and M synoviae, may produce an intensive,
capable of producing a polyarthritis in prolonged and disabling synovitis and ten-
or as it has been called P (plasma)-component of amyloid, is antigenically identical with an a globulin in the blood of
normal individuals (C45, C46, C47).
Changes in serum protein patterns occurred
in human (M145) and experimental amyloidosis (B82, K54, S,7) particularly in
the 7-globulins, and the latter may be
associated with amyloid in the tissues.
General reviews appeared of amyloidosis
that emphasized the complex clinical
manifestations (B33, C87). [These fine reviews provide an excellent introduction to
a dificult field. Ed] A clinical analysis of
42 patients with amyloidosis indicated that
rectal biopsy was the procedure of choice
for establishing the diagnosis (B120). The
postmortem incidence of amyloidosis in
R A (C89) is about 15% and about 4% in
Still's disease (S125), and amyloidosis
seemed to occur on an average about 8
years after joint disease began. Among 316
patients with FMF, 27% had amyloidosis,
and almost all who died succumbed to
renal disease and uremia (G3). Amyloid
deposits in the gastrointestinal tract may be
associated with impaired secretory functions and malabsorption (Bl, B121, F13).
Multiple myeloma, with deposits of
amyloid in the synovial membrane of
joints, may simulate RA (A41).
Mhritis a d Rheumatism, Vol. 13, No. 5 (Septemkr-Octobar 1970)
syndrome, added interest has been stimudinitis in chickens (022).
Another agent, Erysipelothrix insidiosa, lated i n the type of infection and the mode
is also responsible for crippling and de- of pathogenic action of this agent. [See section
forming polyarthritis in swine (S99, S101). on Reiter’t Syndrome.)
T h e polyarthritis
passes successively
A suppurative arthritis caused by a strain
through acute, subacute and chronic stages of streptococcus (S equisimilies) apparentand clinically, the chronic stage was charac- ly caused a serious economic loss i n the
terized by periods of exacerbation and re- swine industry. Infection of swine promission. An ankylosing spondylitis was duced a suppurative polyarthritis with
clearly demonstrated and resulted from the high synovial fluid leukocyte count and
formation of bony plates bridging interver- eventually fairly severe destruction of the
tebral discs. Grossly, after several years of synovium and of the articular cartilage.
chronic infection, there was enlargement of Pathogenesis of breakdown of the articular
the carpal, tarsal and interphalangeal surfaces was studied in this report (R31).
joints and contraction of tendons. Histolog- It was seen that the streptococcal arthritis
ically there was advanced pannus forma- produced a loss of metachromasia of the
tion replacing the articular cartilages, syno- articular cartilage, which was associated
vitis and marked proliferation of the syno- with metaplasia of the chondrocytes and
vial villi. Serologic studies reveal hyper- fibroblasts and by hyperplasia of the cartigammaglobulinemia and hypoalbuminemia. lage cells surrounding vascular canals. T h e
T h e various forms of infectious arthritis hypertrophy and hyperplasia of the cartithat occur i n cattle were classified with lage probably indicated a n attempt, on the
respect to duration, and depending on part of the animal, to strengthen the surtheir pathogenesis, were listed as primary, face against the altered joint environment.
secondary or tertiary (V8). Various pyo- However, instead of strengthening the argenic or nonpyogenic organisms were iso- ticular cartilage, the altered cellular state
lated from synovial effusions of infected appeared to result in altered matrix, which
joints and the characteristics of the synovi- allowed fissuring, erosion and serious dea1 fluid and the synovium were outlined in terioration of the articular cartilages.
considerable detail.
An outbreak of polyarthritis, caused by a ADJUVANT-INDUCED POLYARTHRITIS
virus (Psittacosis-Lymphogranuloma-Vener- Adjuvant disease, on experimentally ineum-Trachoma Group) (PLT) was de- ducible polyarthritis in various strains of
scribed i n recently weaned lambs under 1 rats, is regularly initiated by an intraderyear of age (P57). T h e clinical signs were ma1 injection of complete Freund’s adcharacterized by conjunctivitis, fever, and juvant, or with tubercle bacilli in mineral
lameness due to arthritis. There was a oil alone. A number of studies on this
satisfactory response to treatment with an- experimental model continue to appoar in
tibiotics, especially intramuscular injec- the literature. These studies followed three
tions of oxytetracycline and penicillin. T h e general directions: (a) studies of the
same PTL viral organism was also investi- mechanism or mechanisms of induction:
gated in detail for its disease-producing (b) observations on various protective or
properties in sheep (N27).Since the recent non-protective components and drugs; and
observations that a PLT agent has been (c) evaluation of the inflammatory reisolated from some patients with Reiter’s sponses. T h e adjuvant-induced model has
Arthritis and Rheumatism, Vol. 13, No. 5 (September-October 1970)
been used to screen potential antiinflammatory agents. A study was undertaken to determine the mechanisms whereby adjuvant
disease is suppressed in rats pretreated with
an injection of tubercle bacilli in saline
(G13). I n the earlier literature it had been
noted that newborn animals, treated either
with tubercle bacilli in saline or with complete adjuvant, did not develop adjuvant
disease during the neonatal period (before
5 weeks of age). Moreover they were protected from subsequent induction of arthritis, presumably throughout the remainder
of life. This observation suggested that
immunologic tolerance may be involved.
Animals injected with tubercle bacilli in
saline, interperitoneally, twice weekly for 3
weeks did not develop any disease, although PPD skin tests became positive.
The animals were resistant to the develop
ment of an adjuvant disease induced by the
usual intradermal injection. Various
studies of this phenomenon were undertaken. It was shown that the pretreatment
with tubercle bacilli in saline did not have
any effect on nonspecific inflammatory
lesions produced by histamine and 5-hydroxytryptamine. Moreover, pretreatment
did not affect the ability to induce a significant antibody titer with unrelated antigens
such as bovine 7-globulin, or the induction
of experimental allergic encephalomyelitis.
Moreover, the effect of altering reticuloendothelial (RE) function, either by depressing or altering RE function with methyl
palmitate or enhancing RE function with
zymosan, had no specific effect upon the
subsequent development of adjuvant disease. The transfer of serum to normal
controls from pretreated rats who had developed adjuvant disease did not protect
against adjuvant disease as had been reported before.
It has been previously shown that adjuvant arthritis can be transferred to
imbred healthy donors with large numbers
of sensitized lymphocytes from animals previously injected with adjuvant. However,
in normal animals, presensitized by injections of tubercle bacilli in saline, the recipients of sensitized lymphocytes from pretreated donors did not develop adjuvant
disease, or did so in a very minimal and
scanty form. This inhibitory effect of pretreatment was found to be restricted to the
development of arthritic lesions, since skin
testing with PPD continued to remain positive. The conclusions from this important
study were that the inhibition could be due
to a phenomenon analogous to tolerance:
(a) a decreased capacity of lymphoid cells
to respond to the specific antigen responsible for adjuvant disease, (b) a competing
type of immune response to tubercle bacilli
antigens, or (c) a competing response of
the same type but to different antigens OE
the organism.
I n an additional study (G12), the
phenomenon of antigen competition was
further investigated. It had previously been
shown that when unrelated antigens such
as bovine 7-globulin or bovine serum albumin were incorporated with adjuvant
and then injected into a normal animal,
adjuvant disease was almost uniformly suppressed. On the other hand, this investigation demonstrated that when animals to be
tested were specifically rendered tolerant to
bovine 7-globulin, such animals subsequently injected with a combination of
adjuvant and bovine 7-globulin uniformly
developed polyarthritis.
These data
provide additional evidence for the hypothesis that the inhibition of adjuvant
disease by unrelated protein antigens is due
to the competition of antigens.
Another group undertook the study of
the chemical subfractionation of the tubercle bacilli with an aim toward attempting
to understand what might be the single or
Mhrilb and Rheumatism, Vol. 13, No. 5 (Saptemkr-lktober 1970)
multiple arthritogenic factors in this organism (J28). It was demonstrated, as had
been shown previously, that the Wax D
fraction of tubercle bacilli was highly arthrogenic. Fractions obtained after ultracentrifugation in ether were acetylated
and then chromatographed on silicic acid.
It was found, as had been reported previously by others, that acetylation of the Wax
D apparently blocked the arthritisinducing ability of that compound. Moreover, the acetylated subfractions did not
induce arthritis. Another interesting observation was that rats injected with acetylated Wax D, although not developing arthritis, were subsequently protected against
the arthritis-inducing ability of unaltered
(unacetylated) Wax D.
A detailed study was also made on the
influence of lymphatic tissue on the development of adjuvant-induced arthritis in
rats (R76). In this experiment, adjuvant
was injected intradermally into one hind
foot. It was demonstrated that prior (5
days), simultaneous with, or 5 days postinjection removal of the regional lymph
nodes draining that hind limb almost completely inhibited the polyarthritis. On the
other hand, when lymph nodes were removed from the contralateral side, there
was no inhibitory effect, nor did splenectomy or thymectomy have any properties of
diminishing the time of appearance or the
severity of arthritis. [These studies appear
to confirm the previous speculations that
the adjuvant disease is due to a delayed
hypersensitivity reaction which develops
parallel to the development of immunologically competent cells. It is possible that
these cells could be acting, in the pathogenesis of adjuvant-induced polyarthritis, against their own connective tissue antigens, or, as an alternative, against fragments
of the tubercle bacilli that are disseminated into the peripheral tissues. Ed]
These authors also make mention of the
interesting observation that thymectomy
performed in newborn rats prevented them
from developing polyarthritis in adult life.
On the other hand, thymectomy performed
on rats 5 weeks of age had no inhibitory
effect on adjuvant-induced polyarthritis.
Some of the inflammatory mechanisms
and other derangements that occur after
adjuvant injection into the rat were
studied (B102, G60, W42). Of the various
inflammatory parameters that were followed, a previously described test called
“plasma inflammation units” was the most
reliable. In addition, the sedimentation
rate, serum glycoproteins and copper, and
the size of reactive lymph nodes were all
increased markedly in the adjuvantinjected animals. Most of these tests could
be reversed quite promptly (including r a p
id diminution in size of lymph nodes) after
treatment with dexamethasone.
A specific
glycoprotein appeared during the course of or immediately after
adjuvant injection (B102). Specificity of
serum concentration of this protein component was clearly defined by the immunologic procedures utilized in this study. In
another study (W42), it was concluded
that, although serum changes paralleled to
a certain extent the degree of pathologic
activity, most reactions that were studied in
this series were found to be nonspecific
and, in general, reflected a common systemic response to inflammation. In another
study, the anemia of adjuvant-induced arthritis in rats was likened to the chronic
anemia that develops in man in RA and
other chronic illnesses (L89). Therefore,
this experimental animal model was considered to be a valuable one in which to
study the anemia of chronic disease states.
I n a pathologic study (M65), a previously unnoticed or unstudied joint, namely,
the temporomandibular joint of the rat,
Arthritis and Rheumatism, Vol. 13, No. 5 (September-October 1970)
was found to be involved in adjuvantinduced arthritis. T h e sequential histologic
studies from the first 24 hours after injection and up to 300 days were reported in
some detail.
The effects of various therapeutic agents
were studied, in more or less random fashion, for effects upon adjuvant-induced arthritis. A new cytotoxic compound of the
anthracycline group, rubidomycine, seemed
to prevent or to markedly inhibit arthritis.
Rubidomycine was given from Day 4 to
Day 9 after adjuvant injection and its effectiveness appeared to be clearly demonstrated. For comparison, tetracycline had no
inhibitory effects (Ql).
Statolon, a derivative of the mold Penicillium stoloniferum, whose activity is believed to be due to a virus-like particle that
infects the mold, was found to also inhibit
the development of adjuvant-arthritis (18).
Since Statolon is an effective inducer of
interferon, it has been suggested by the
authors that a virus is involved in the
pathogenesis of adjuvant-arthritis, and that
such a virus has some relationship to lymphoid cells. The authors also demonstrated
that the PPD reaction, usually a prototype
of cellular hypersensitivity, was not inhibited in the animals given adjuvant and
Statolon. [More recently the same authors
have demonstrated that Pyran copolymer, a
totally synthetic polyanion, which is also
capable of inducing interferon formation,
will likewise suppress adjuvant-induced arthritis (K19). Ed]
Apparently adjuvant arthritis is also partially inhibited by the administration of
the hormone, estrone. This was apparently
not mediated through either the adrenal or
the ovary, since surgical removal of either
or both of these glands did not block the
inhibitory effect of estrone (T8.5). Gold
salts in the form of myochrysine, when
given daily for 14 days prior to adjuvant
injection and 18 days thereafter, had no
inhibitory effect on the time of appearance
or the severity of adjuvant disease (525).
When cysteine was given simultaneously
with or subsequent to the administration of
adjuvant, it had an inhibitory effect upon
the appearance of arthritis (Dl). The authors speculated that the effect of cysteine
is much the same as that of D, Z-penicillamine, which can apparently cause a decrease
in the immunologic response against a simultaneously administered antigen. They
also speculated that the process of protein
synthesis in the developing lymphoid cells
may be inhibited. In another study, by
these same authors (R77), the effect of
certain neurohormones (including adrenaline) and atropine were studied for their
effect on cellular responses occurring in
lymph nodes in animals given adjuvant
injections. They described a diminution oE
the features of polyarthritis and some
change in the ratio of lymphoblastic and
plasmocytoid cells. Other so-called neurohormones, such as gynergen and acetylcholine, did not have a similar effect.
T h e authors tentatively concluded that the
effect of these hormones probably operates
through the hypothalamohypophyseal- adrenocortical axis.
Aleutian disease afflicts multiple strains
of mink. I t seems to be unusually severe in
animals homozygous for the Aleutian gene
(aa), hence the origin of the name. The
disease is characterized by extensive proliferation of plasma cells, hypergammaglobulinemia, a progressively developing renal
disease including severe glomerulonephritis, infiltrative plasma cell hepatitis and
hyaline or fibrinoid vascular lesions. It is
quite clear that the disease syndrome is
transmissible by cell-free filtrates and is
due to the presence of a virus-like infec-
M h i t i s and Rheumatism, Vol. 13, No. 5 (September-Oetober 1970)
tion. I t was noted in the early studies of
this disease that profound hypergammaglobulinemia was present in some infected animals. A myeloma-like monoclonal
y G-immunoglobulin component was noted
to appear in the sera of 10-15% of diseased mink that survived and harbored
the infection for periods longer than 12
months (P71). The presence of these
serum protein abnormalities paralleled
quite closely the striking histologic lesions
of a progressive glomerulonephritis in
mink (W75, W76). These renal and other
vascular lesions had great similarity to
those found in human periarteritis or
Specific antisera to mink y-globulins,
when reacted with an infectious mink
serum pool, markedly reduced the infectivity of such sera suggesting that the infectious agent of Aleutian mink disease circulates in a complex with host 7-globulins
The renal glomerular lesions were
studied in detail (H58,H59). By light
microscopy the earliest and most constant
lesion was glomerular thickening and the
accumulation of eosinophilic material in
the mesangial areas. Increased glomerular
size and cellularity, and the presence of
inflammatory cells were also seen. The
most frequent and early ultrastructural
changes were deposition of electron-dense
granular material in the subendothelial
mesangium as well as a mesangial cellular
and matrical increase. Basement membrane
irregularity and subepithelial and intramembranous dense deposits were also
present. The glomerular lesions in Aleutian disease would appear to represent an
example of the proliferative glomerulopathy that can be initiated by a filtrable viral
The exact nature of the macromolecular
deposits is not quite clear. However, recent
work (H58) utilizing the fluorescent antibody technic showed that early in the
course of Aleutian disease there is deposition of 7-globulin and complement Ule)
principally along the basement membrane.
Fibrin and albumin were not detected until late in the disease, if at all, and probably
represented secondary changes. I n the diseased glomeruli, a definite nodular pattern
of fluorescence was noted. I t seemed likely
that the fluorescent material corresponded
to the deposits visualized with the electron
I t was noted (H59) that mink homozygous dominant (AA) or heterozygous (Aa)
for the Aleutian gene have a slower clinical
course with slower development of glomerular lesions than homozygous recessive
mink (aa). Whether the rate of development of the glomerular lesions in AA or Aa
mink relates to variable formation of macromolecular substances, or to variable
ability of the cells to degrade the deposited
macromolecular material, or both, is not
known. If there is formation of less macromolecular material in AA or Aa mink, it
may be due to a lower virus titer in these
genotypes. I t was reported that virus could
be detected at any time until death from aa
mink, whereas virus was detected from Aa
mink only from the second to the twelfth
weeks postinoculation (E17). On the other
hand, the possibility that aa mink do not
degrade macromolecular material deposited in the kidney as efficiently as AA or
Aa mink may relate to the presence of
abnormal lysosomes in aa mink. All aa
mink have abnormal membrane-bound
structures in many cells of the body, some
of which were thought to be lysosomes, the
hallmark of the Chediak-Higashi syndrome
(P3). I t now appears that aa mink and
man and cattle with Chediak-Higashi syndrome are unable to handle bacteria (and
presumably viruses) as efficiently as AA or
Arthritis and Rheumatism, Vol. 13, No. 5 (September-October 1970)
Aa mink.
Seven of 273 mink examined (83 with
clinical Aleutian disease) demonstrated increased levels of serum a,-globulin. The
kidneys in these mink were severely damaged with proteinaceous deposits in the
glomeruli and marked lymphocytic and
plasmacytic reaction in the interstitial and
periglomerular areas. Periodic acid-Schiff
reactions and the results of Congo red
staining indicated that the proteinaceous
material was amyloid (K54). I n all of these
animals, the protein contributing to the
increased az-globulin level made up from
3 0 4 0 % of the total serum protein and
was characterized as a macroglobulin (19s)
containing variable amounts of carbohydrate, generally in the range of 13%.
[Marked increase in ,,-macroglobulins is
associated with many clinical and experimental iiephrotic states. Ed]
The spontaneously occurring disease
which develops in the NZB (New Zealand
black) strain of mice, or in some of their
hybrids, has been subjected to intensive
investigation in recent years. Several
features of this disease, hemolytic anemia,
antinuclear antibodies and glomerulonephritis, closely resemble those of human
Mice of the NZB strain appear normal at
birth, but between 4 and 9 months of age,
the majority develop a hemolytic anemia,
analogous to human autoimmune hemolytic anemia of the warm antibody type (B24,
B25, H78). T h e first abnormality detected
is a positive direct antiglobulin test eventually positive in practically 1 0 0 ~ oof the
mice. The hemoglobin and hematocrit fall,
reticulocytes increase, and splenomegaly
develops. I n addition, plasma cells become
prominent in the spleen and lymph nodes,
and also in the lungs, kidneys and thymus.
Arthritis and Rheumatism, Vol. 13,
The presence of germinal centers in the
thymus has been noted, but their relationship to intrinsic thymic tissue is in some
doubt (D25). In the kidneys, glomerular
lesions, with tubular casts, develop in more
than 50% of the females and in about 25y0
of the males. Also about 20y0 of the mice
develop antinuclear autoantibodies (H78).
The results of crossing the NZB strain
with other inbred mouse strains show that
the autoimmune character of the NZB
mouse is expressed in its F, hybrids, although manifested in different ways. For
instance, it has been found that hemolytic
anemia developed in 1 0 0 ~ oof F1 hybrids
from NZB x NZC matings, independently
of the sex of the NZB parent.
Much of the current investigation has
been divided into four major categories:
(a) Study of the genetic features
(b) Additional evaluations of the immunologic characteristics of the disease
(c) Further investigation of the structural abnormalities, including ultrastructural features
(d) Significance of the finding by several
groups of virus-like particles in various
organ tissues and the relationship, if any,
of that finding to the spontaneous develop
ment of leukemia in some of the mice
If, as seems likely, an infectious component is involved in the disease process
in mice, it seems quite probable that genetic factors also play a role in allowing the
NZB mouse and its hybrids to express or
develop various features of the disease. In
one study (B125), the genetic features and
natural history of several New Zealand
mouse strains and their hybrids were carefully studied. The natural histories of the
NZB, NZY, NZW and NZB/NZW animals.
were described. Features of autoimmune
disease were found in the B, Y, W and
B/W strains. Renal disease was transmitted
independently of antinuclear factor and
No. 5 (September-October 1970)
Coombs’ positivity and appeared to be determined by multiple genes. The genetic
contribution from the NZW mate appeared
to favor development of ANF in addition
to Coombs’ positivity. Segregation of these
genetic factors in the F, and backcross
produces a proportion of mice in whom
ANF and Coombs’ positivity do not develop. I n this study, it was stated that it
was not possible to transmit autoimmune
disease from NZB mice to Swiss albino ICR
animals with cell-free filtrates of tissuehomogenates. [In another study reported
in the last Review, purified cellfree filtrates
of splenic tissue obtained from old NZB
mice were reported to be able to induce
mild renal and hemolytic disease (including a positive Coombs’ test) in the Swiss
variety of mice (M92). Ed]
The immunologic and genetic aspects of
this interesting mouse model have been
investigated in considerable detail. Thus,
in a study of autoimmune disease in the F1
hybrid (NZB x CFW) (B24), the animals
were shown to have a low and delayed
incidence of positive direct Coombs’ tests,
whereas the incidence of serum antinuclear
factor in this cross was markedly increased
by comparison with the parental NZB and
CFW strains. There was no particular correlation in the Fl hybrids between the
appearance of renal disease and the autoantibody state or the degree of thymic
proliferation. I n this cross-strain there was
a very high incidence of thymoma superimposed on a previously nonmalignant proliferative lesion of the thymus.
In one study utilizing NZB mice, as well
as a controlled strain of mice (E48), both
groups were injected with sheep red blood
cells to test for immunologic competence.
When the response was expressed as
plaque-forming cells (PFC) per million viable cells, the response curve of the spleens
of the baby NZB mice was very similar to
the response in the spleens of the adults.
Moreover, the response in the lymph nodes
of the babies was as high and more sustained than that of the adults. In the
control strain of baby mice at the same age,
the response in the spleen and the lymph
nodes was reduced and delayed compared
with adults. Neither strain gave a significant response in the thymus.
In another study (B124), it was found
that delayed hypersensitivity to hen egg
albumin, BSA and PPD developed normally in NZB/NZW mice, when their response
was compared to responses in a control
C,H strain of mice. Moreover, relative
tolerance to the bovine 7-globulin was produced in adult NZB/NZW and NZB mice.
Also, in this hybrid strain, hydralazine and
mesantoin did not accelerate or aggravate
autoimmune disease. When parabiosis was
induced between old NZB/NZW and
young animals of this hybrid strain, or to
NZW or ICR mice, none of the parabionts
developed renal disease. Nor did i t seem
to accelerate the onset of autoimmune disease in the young NZB/NZW animals despite the fact that in some of the parabionts, antinuclear factor could be demonstrated for from 4 to 8 weeks after
the parabiotic union had been interrupted.
[It seems possible, in view of more recent
findings, that these animals were not observed for a suficiently long period of time
to state unequivocally that various manifestations of autoimmune disease would
eventually not develop. Ed]
The young NZB mice and the (NZB X
CBA) F1 hybrid mice gave an enhanced
immune response to a soluble protein antigen (alum-precipitated BSA) fw45). Evidence was also found that it was more
difficult to render some of the NZB animals
tolerant by pretreatment with centrifuged
material (which presumably contained aggregate-free BSA). These results are consist-
Arthritis and Rheumatism, Vol. 13, No. 5 (September-October 1970)
ent with other previously derived evidence
that these mice are immunologically hyperactive.
T h e NZB/NZW Fl hybrid strain of mice
developed renal lesions which resemble
closely lupus nephritis. A detailed study of
the developing renal lesions from age 1
month was made in this hybrid strain
(M82). By the age of 1 month the mice
showed glomerular deposits of electrondense material, which, with age, became
more extensive on the intracapillary and
epithelial aspects of the capillary basal
membrane. These features were visible first
by electron microscopy and could b? seon
before the proliferative and membranous
changes were visible by light microscopy.
Moreover, electron microscopy detected
such changes before significant immunoglobulin accumulations could be detected
by immunofluorescence. T h e nephritis a p
peared to be more rapidly progressive in
the female and led to obliteration of efferent capillary loops by intracapillary cells
and electron dense material. Later, there
was widespread exudation and the accumulation of all plasma proteins.
Immunofluorescence of the renal lesion
(M81) revealed the presence of many plasma proteins, particularly immunoglobulins
in the early stages of the glomeruli. Later
on, albumin and fibrinogen were present in
the glomerular lesions. T h e exudates had
the ability to bind guinea pig complement,
which suggests that they include antigenantibody complexes (L9). T h e development of glomerulonephritis in NZB/NZW
mice was closely related to the formation of
antinuclear, particularly DNA antibodies.
The developing inflammatory glomerular
lesions were characterized by the deposition of IgG, PIC and DNA and possibly
other nuclear antigens. Presumably, all of
these molecules existed as complexes in a
granular pattern along the capillary walls
and in the mesangia. Enhancement of the
antinuclear antibody response by active immunization of young NZB/NZW mice with
DNA-methylated BSA hastened the development and increased the severity of the
glomerulonephritis. Similarly, injections of
soluble DNA into these mice, who also
harbored circulating DNA antibodies,
caused rapid progresssion of the renal disease.
Probably the most significant observations in regard to the pathogenesis of autoimmune disease in mice have been the
observations, made almost simultaneously
from several widely separated laboratories,
of the presence of virus particles, as observed by electron microscopy, and identifiable as the Type C murine leukemia virus
(E2, M93, P85, Y6,Y7). I n one study (Y7),
the kidneys of 22 of 24 mice (of which 14
had lymphoid leukemia) showed slight or
extensive glomerular changes typical of autoimmune disease. Electron microscopy of
the kidney tissues revealed generalized increase in the thickness of the basement
membrane as the most characteristic lesion.
A large number of mature and immature
Type C murine leukemia virus particles
were observed in the kidneys of 21 out of
24 of the NZB mice. Budding of virus
particles was observed at the plasma membrane of the visceral and parietal epithelium of the glomeruli, from the epithelium
of the proximal and distal convulated tubules, and in smooth muscle cells and renal
arterial walls.
Virus particles were observed in great
numbers in kidneys with changes characteristic of autoimmune disease, regardless of
the extent of infiltration by leukemic cells.
T h e number of virus particles could be
correlated with the severity of the kidney
lesions in animals with spontaneous autoimmune disease in the kidneys. Tissues of
apparently normal kidneys of I-month-old
NZB strain mice revealed only a few Type
Arthritis and Rheumatism, Vol. 13, No. 5 (September-October 1970)
C virus particles. On the other hand, an
increasing number of virus particles were
found in the kidneys of 3-month-old mice
of the same strain showing very slight glomerular changes.
An electron microscopic study of sections
of lymph nodes, spleen, liver, bone marrow, thymus gland and lungs of the same
leukemic mice demonstrated immature and
mature Type C murine virus particles.
These studies also revealed (Y6) Type C
particles in the parenchymatous cells of the
liver and kidneys.
Similar particles were found in many
tissues (spleen, thymus, lymph nodes, bone
marrow and pancreas) in both normal and
thymectomized mice (P86). Mice of the
NZB strain which had been introduced
into a germ-free environment by cesarean
section also demonstrated Type C virus
particles in many of the tissues as described
above (E2, P85).
I n germ-free mice, Coombs’ positivity
developed between the sixth and eighth
month, and these reactions increased in
intensity with age. The development of
Coombs’ positivity in the germ-free animals
was delayed slightly when compared with
conventional mice of the same strain. Two
out of three 8- to 11-month-old mice and 1
of four 6-month-old mice were antinuclearantibody positive.
Although other strains of mice reared in
germ-free climate were reported to have
very low serum-immunoglobulin levels, the
germ-free NZB mice had immunoelectrophoretic patterns nearly identical with patterns of conventional mice (E2).
Swiss mice were given neonatal intraperitoneal inoculations of cell-free filtrates
prepared from the spleen of old NZB mice
that had large quantities of Type C murine-type particles in the splenic cells. Lymphoid cell and plasma cell hyperplasia as
well as hypergammaglobulinemia de630
veloped in some of the inoculated Swiss
mice. Moreover a mild hemolytic disease,
positive indirect Coombs’ test and renal
disease with proteinuria and glomerular
lesions were observed. Control observations
in which NZB lymphoma filtrates were
injected intraperitoneally into adult Swiss
mice gave negative results. I n the neonatally-injected Swiss mice, Type C virus-like
particles were shown by electron microscopy to be present in the distinctive locations,
notably in the basal foldings of the convoluted tubules and other typical sites. Three
of 28 neonatal Swiss mice inoculated with
NZB cell-free filtrate developed malignant
lymphoma and a leukemic blood picture at
7 months. The incidence of lymphoma was
about 10 times that of spontaneous lymphoma in 100 control Swiss mice (h193).
The presence of a virus-like agent could
possibly explain the observation (M51)
that an autoimmune hemolytic anemia was
produced in mice of 4 different strains that
were thymectomized neonatally and then
implanted (intraperitoneally) with Millipore diffusion chambers containing thymic tissue from newborn mice of the NZB
strain. Of a total of 60 surviving animals,
50 developed a positive Coombs’ test and
other evidences of autoimmune hemolytic
anemia. On the other hand, it could not be
completely ruled out that thymus tissue
from the NZB animals, which continued to
grow in the Millipore chambers, may possibly have elaborated a humoral factor
which could contribute to the development
of the hemolytic phenomenon. [Zn view of
the recent observations on virus particles,
the latter possibility seems unlikely. Ed]
A number of therapeutic trials were undertaken in NZB autoimmune disease.
These studies may have relevance to human SLE (R74). Cyclophosphamide,
when given continuously from 120, 160 or
200 days of age, decreased the incidence of
Arthritis and Rheumatism, Vol. 13, No. 5 (September-October 1970)
severe renal disease from 100 to 6.77’ in
female NZB/NZW mice. Cessation of
therapy at one year of age was not associated with an increase in the incidence of
renal disease. Mice given long-term treatment lived twice as long as untreated mice.
The cyclophosphamide dose in these animals was 1.8 mg/mouse/week. I n mice given short courses of cyclophosphamide in a
dose of 5 mg/mouse/week for 4 weeks,
there was also an increase in longevity,
although the effect was not as marked as
with long-term treatment.
On the other hand, when 6-mercaptopurine therapy was given in NZB mice, shortly
before the expected onset of autoimmune
hemolytic anemia, the treatment did not
delay or modify subsequent development of
Coombs’ antibodies (C35).
When azathioprine was administered to
NZB/NZW mice, the appearance of
nephritis was suppressed. However, malignant lymphoid neoplasma, particularly
affecting the thymus, occurred in 7 out of
12 of the mice so treated (C36). Similar
neoplasms developed in 6 out of 8 young
NZB mice also treated with azathioprine.
No such tumors occurred in a similar number of control animals during the period of
observation. Such a situation could possibly
be explained, as can radiation-induced leukemia, by the fact that the cytotoxic drug
can interfere with the natural defenses
against tumor development of viruses. [If
this result is confirmed by additional
studies, it has important implications in
the use of cytotoxic drugs in SLE and
related autoimmune states. Ed] (See section
on Cytotoxic Therapy.)
T h e appearance of nephritis in
NZB/NZW hybrid mice was interrupted
and the severity and activity of the renal
lesions reduced by treatment with the corticosteroid drug, betamethasone phosphate
(C37). Since the renal lesions in NZB/
NZW mice often contained large quantities of fibrin, even in the early stages,
animals were treated with the anticoagulant, warfarin. The clotting time was prolonged fourfold, but this therapy did not
appear to influence the progress of the
renal disease (M80). In another study
(S186), when antithymocyte globulin was
administered to NZB/NZW hybrid mice,
the treatment had a deleterious effect on
survival but no substantial effect on the
renal histopathologic changes. It appeared
to these authors that the decreased survival
rate in the treated animals was related to
an exacerbation of an underlying viral
A systematic serologic study was made in
17 purebred strains of mice and 2 random
outbred strains of mice that had been
maintained for a long period of time under
pathogen-free, but not germ-free, conditions (B23). Direct Coombs’ tests were negative in all of the strains examined with
the exception of the NZB strain. On the
other hand, it was clearly shown that the
NZB strain of mice is not the only one to
possess antinuclear factors, and several
strains were described where the incidence
of ANF was greater than in the NZB
mouse. Of the 19 strains of mice that were
investigated, 3 pure line strains had a high
incidence of the factor; these were the A,G,
DBA/2 and C57 BR. Incidence of ANF in
these strains, respectively, were 27.8, 26.5
and 51.1%, compared to 25% positive tests
in the NZB strain. It was also shown that
the incidence of ANF was somewhat lower
in those mouse strains that were
maintained in conventional housing quarters.
It was thus demonstrated that both genetic factors and environmental factors play
roles in determining the presence or absence of ANF in the serum, and possibly in
the development of lesions.
Arthritis and Rheumatism, Vol. 13, No. 5 (September-October 1970)
A rheumatoid-like arthritis was said to
have been produced in swine that were
given whole blood transfusions from swine
with E insidiosa arthritis (S100). These
animals received 20 ml of whole blood
‘twice a week for one year. They developed
carpal and tarsal joint synovitis with pannus formation. On the other hand, animals
that were given serum from the same arthritic swine at the same dosage level and
frequency for one year did not develop any
alteration in the synovial fluid or synovial
membrane. All bacteriologic cultures made
from the blood and serum of both arthritic
and normal donor swine, and from all
recipient swine were said to be negative. It
was therefore concluded that there was a
toxic and antigenic substance in the blood
of the arthritic donor swine, since the normal swine given the whole blood transfusions developed arthritis. Also the toxic
antigenic factor appeared to be related to
the cellular rather than serum components
of the blood.
An arthritis was induced in rats by injection of the exudate formed in granuloma
pouches (F6). The exudate was injected
under the skin of the rat’s foot and swelling
of the limb followed within a matter of
minutes. This swelling was apparently
ascribed to the presence of a histamine or
H-substance, and it was preventable, at
least in part, by the prior intraperitoneal
injection of the antihistamine, phenergan.
Based upon studies reported previously,
rabbits were repeatedly injected with
formalin-killed E coli until eventually the
serum of these rabbits contained moderately high titers of a rheumatoid factor-like
substance (RFLS). To test the hypothesis
that joint inflammation in RA resulted
from leukocyte response to the interaction
within the joint of 7-globulin and rheuma632
toid factor, autologous y-globulin was injected into the knees of rabbits containing
RFLS in their serum (78). A local nonsustained synovitis was produced regardless of
whether RFLS was present or absent in
the serum.
A symposium on equine bone and joint
disease was published (S205).
It was reported previously that a chronic
arthritis could be induced by repeated injections of streptolysins (SLS) into the
joints of rabbits. T h e SLS was said to be
nonantigenic and to have as its main role
of action the damage of lysosomal membranes with subsequent joint damage
caused by release of hydrolytic enzymes.
Moreover, in that initial report (W49),
SLS was said to be the only factor within
the streptococcal organism to have such an
effect on the synovial membrane.
I n a more recent report (G22), streptococcal extracellular products (SEP), which
did not contain SLS, were produced by
sonication of Group A streptococci. When
SEP was injected into the joints of rabbits a
severe synovitis developed promptly. If, on
the other hand, SEP was heated to 100°C
for 30 minutes prior to injection into the
joint, an inflammatory reaction did not
develop. It therefore appeared that a heatlabile product, present in the SEP, was
capable of producing intense sterile arthritis when injected locally.
Repeated intra-articular injections of
SLS into the knee joints of rabbits at
weekly intervals provoked a chronic arthritis (C98), much like that described previously. The authors were not able to find
complemen t-fixing antibodies to subcellular particles, or inhibition of the release of
enzymes by sera of rabbits that had been
repeatedly injected.
Moreover, in experiments in vitro these
Arthritis and Rheumatism, Vol. 13,
No. 5 (September-October 1970)
authors were not able to demonstrate the C-polysaccharide can act as durable toxic
release of enzymes from lysosomes by SLS, materials, which, at least, play a role in this
although it was shown that appreciable model (S53).
amounts of B-glucuronidase were released
In another report (W50), a number of
from a heavy granular fraction that con- polyene antibiotics resembled SLS in their
tained lysosomes and also mitochondria. membrane-disrupting capabilities. Such anThe authors of this study found very little tibiotics were previously noted to be capaevidence that the joint damage was caused ble of killing fungi by virtue of their capacby release of hydrolytic enzymes. Cartilage, ity to disrupt the membranes of susceptible
when it was destroyed, appeared to be cells. The antibiotics included filipin,
removed by the encroachment of pannus, etruscomycine, pimaricine and amphoteriand although this may have been caused in cin B. There was evidence that sterols
part by enzymes at the pannus-cartilage within cell membranes were the unique
junction, the remaining cartilage matrix component of cell membranes with which
did not seem to lose its metachromasia. the polyenes reacted. Although the polyMoreover, these authors found that fibrin enes were hemolytic and capable of disruptpersisted in large amounts in these joints. ing lysosomes, the function of mitochonHad a release of acid hydrolases from poly- .dria was unaffected. When polyene antimorphs and synovial cells been an impor- biotics were injected repeatedly into the
tant factor, it is reasonable to have expect- knee joints of rabbits, acute and chronic ared that rapid removal of fibrin as well as thritis resulted and their order of activity
loss of metachromasia in the cartilage was identical to that noted in their effect
on isolated lysosomes and on artificial
would have occurred.
A single injection of isolated fragments membranes. The features of the lesions proof Group A streptococcal cell walls into the duced resembled those resulting from rejoints of rabbits stimulated an initial acute peated injections of SLS. But changes in
reaction which was followed by a pro- articular cartilage predominated: loss of
longed inflammatory response. The chronic matrix substance, fibrillation of the cartiprocess was characterized by hyperplasia of lage surface, formation of vertical clefts
the synovial cells, diffuse infiltration of the and fissures in the substance of cartilage and
villi by macrophages and focal coIlections proliferation of the chondrocytes to form
of lymphocytes in the stroma of the villi. chondrocyte clusters. The pathologic
These histologic changes were quite similar change, therefore, was somewhat closer to
to those seen in the early stages of RA. human degenerative joint disease than RA.
The most potent of the plyenes was
Antibodies, specific for the mucopeptide
and groupspecific C polysaccharide anti- filipin, and the least potent was amphoterigens of Group A streptococcal cell walls, cin B. Cortisone, when given intra.were labeled with either fluorescein or with articularly shortly before filipin, diminished the subsequent acute changes pro1261. These were used to demonstrate the
presence of the respective antigens in the duced by the polyene. T h e results of these
synovial tissues. I t was remarkable to ob- experiments were compatible with the hyserve that these antigens persisted within pothesis that agents that disrupt natural
the macrophages, after a single intra- and artifical membranes are capable of
articular injection, for at least 5 weeks and inducing inflammation via the release of
indicated that the mucopeptide and the lysosomal enzymes.
Arthritis and Rheumatism, Vol. 13,
No. 5 (September-October 1970)
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