Arthritis in DBA/1 Mice Induced with Passively Transferred Type II Collagen Immune Serum

Arthritis was induced in DBA/1 mice by passive transfer of syngeneic anti-type II collagen (CII) serum concentrate. After transfer of serum containing 0.2 or 0.5 mg anti-CII auto-antibodies the first clinical signs of arthritis appeared 48 h after injection. Severe clinical arthritis was detected 96 h after injection. Immunohistochemical analyses of joints 48 h after serum injection revealed synovial foci in intercarpal and metacarpophalangeal joints of macrophage-like cells, expressing C3bi-receptors and major histocompatibility complex class II molecules, and infiltration of few CD4+ lymphocytes. Later (96 h after injection), the inflamed synovia were dominated by C3bi-receptor+ polymorphonuclear cells. In contrast to conventionally induced collagen arthritis (CIA), the inflammatory infiltrates, filling joint spaces and synovial tissue, were extensively dominated by polymorphonuclear cells, whereas macrophage-like cells expressing class II molecules and a few T cells were seen only in the periphery of the developing pannus. The anti-CII serum induced arthritis may be used as a model for studies of humoral mediated mechanisms operating in conventionally induced CIA as well as in rheumatoid arthritis.     

Importance of the intestinal inflammatory reaction in salmonella-mediated intestinal secretion.

The ability of Salmonella typhimurium to invade the intestinal epithelium is essential to the pathogenesis of salmonella-induced intestinal secretion. This invasion is accompanied by an intense acute inflammatory reaction. The present study tests the hypothesis that the acute inflammatory reaction may have a role in the pathogenesis of salmonella-induced secretion. Two groups of rabbits infected with S. typhimurium were studied: normal animals and animals pretreated with nitrogen mustard. Nitrogen mustard depletes the polymorphonuclear leukocyte pool and thereby prevents the formation of an acute inflammatory reaction. In vivo ligated ileal loops were constructed and infected 72 h after nitrogen mustard administration when polymorphonuclear leukocytes were undetectable. Nitrogen mustard treatment markedly inhibited salmonella-induced secretion. Ileal histology in normal animals infected with S. typhimurium revealed an intense acute inflammatory reaction, while in animals pretreated with nitrogen mustard only a rare polymorphonuclear leukocyte was seen. The antisecretory effect of nitrogen mustard was not merely a nonspecific effect since nitrogen mustard treatment did not inhibit cholera toxin-induced secretion and did not alter either ileal morphology nor the activities of various intestinal enzymes in normal animals. Nitrogen mustard also did not alter the virulence of the inoculated S. typhimurium. These data suggest that the mucosal inflammatory reaction induced by salmonella invasion may be important to the pathogenesis of the salmonella secretory process. The mechanism by which the inflammatory reaction stimulates secretion is not known.     

The immune response to pertussis in the 6-day air pouch: a model of chronic synovitis.

We have developed a model of prolonged immunological inflammation in the rat which has a structural resemblance to the synovial changes in rheumatoid arthritis. Pertussis vaccine was injected into 6-day-old subcutaneous air pouches in animals previously sensitized with pertussis vaccine. The resulting inflammatory response persisted up to 30 days. Examination of exudates showed a wave of polymorphonuclear leucocytes over a 13-day period followed by a mononuclear cell predominance up to 30 days. Histologically, an early polymorphonuclear cell infiltration was followed by the formation of a lining layer of large eosinophilic mononuclear cells, together with deep collections of lymphocytes and plasma cells. Concentrations of the acute-phase reactant alpha 1-glycoprotein, in both serum and exudate, peaked at 3 days. This suggests that the local production of interleukin I in this type of tissue reaction is more closely related to the acute inflammatory phase than to more chronic interactions between monocyte derived cells and lymphocytes.     

The immune response to pertussis in the 6-day air pouch: a model of chronic synovitis

We have developed a model of prolonged immunological inflammation in the rat which has a structural resemblance to the synovial changes in rheumatoid arthritis. Pertussis vaccine was injected into 6-day-old subcutaneous air pouches in animals previously sensitized with pertussis vaccine. The resulting inflammatory response persisted up to 30 days. Examination of exudates showed a wave of polymorphonuclear leucocytes over a 13-day period followed by a mononuclear cell predominance up to 30 days. Histologically, an early polymorphonuclear cell infiltration was followed by the formation of a lining layer of large eosinophilic mononuclear cells, together with deep collections of lymphocytes and plasma cells. Concentrations of the acute-phase reactant alpha 1-glycoprotein, in both serum and exudate, peaked at 3 days. This suggests that the local production of interleukin I in this type of tissue reaction is more closely related to the acute inflammatory phase than to more chronic interactions between monocyte derived cells and lymphocytes.     

Studies on fibronectin in inflammatory vs non-inflammatory polymorphonuclear leucocytes of patients with rheumatoid arthritis. I. Immunofluorescent and flow cytometric analysis.

Using indirect immunofluorescence and flow cytometry, we studied the reactivity of an antibody to human fibronectin with human polymorphonuclear leucocytes (PMNL). Our main objective was to compare the intensity of reaction of this antibody with inflammatory vs non-inflammatory PMNL. We used peripheral blood PMNL as a source of non-inflammatory cells and PMNL isolated from the synovial fluid of patients with rheumatoid arthritis as a source of inflammatory cells. Our findings revealed considerably brighter staining of the inflammatory PMNL. Using flow cytometry as a method of measurement, a difference in fluorescence intensity of at least 40 channels (log scale) was observed in all 12 patients studied when comparing peripheral blood with synovial fluid PMNL. In inflammatory PMNL, fibronectin was found both at the intracellular and membrane levels of the cell whereas fibronectin could be detected only intracellularly in non-inflammatory PMNL.     

Needle biopsy in gout and pseudogout (author's transl)]

Introduction: Radiological and morphological findings in advanced arthritis urica and pyrophosphate arthropathy are well known. In contrast, the early changes of synovial membrane in these disturbances of metabolism pose diagnostic problems. With the assistance of various cytological techniques and polarizing microscopical as well as electron microscopical investigation it was examined to what extent needle biopsies can be helpful in the differential diagnosis of gout and pseudogout. Material and Methods: In 8 patients with gout and 11 patients with pseudogout synovial fluid and small tissue specimens could be obtained with the aid of the Parker-Pearson needle. Both fluid and tissue specimens were investigated light and electron microscopically. Cell counts were evaluated in a Rosenthal chamber. The differentiation of the cells in stained smears was done by counting 200-600 cells per case. Crystals were identified by polarizing microscopy in wet preparations of freshly aspirated synovial fluid. Results: Polarizing microscopy of synovial fluid detected intra- as well as extracellular urate and pyrophosphate crystals. The wedge-shaped urate crystals and the larger partly polygonal pyrophosphate crystals showed different polarizing microscopical properties and a negative birefringence. The absolute cell counts in gout were higher than those in pseudogout. The relative cell counts of the different cell types in synovial fluid showed more variation in gout than in pseudogout. Cases with acute gout developed a relative leukocytosis in contrast to a relative lymphocytosis in chronic gout. A relative leukocytosis was constant in all patients with pseudogout. Sclerosed areas with scarce and plump villi as well as sometimes hyperplastic and polymorphous synovial cell layers could be demonstrated histologically in the tissue specimens of the needle biopsies in cases with gout. Urate crystals were less frequent in specimens fixed in formalin. The histological alterations in pseudogout were uniform, 2-4 rows of slightly pleomorphic synovial cells lined the inner surface of the joint capsule, sclerosing alterations were less frequent. Pyrophosphate crystals and calcified particles were seen within the synovial lining cells, the connective tissue and the enodthelial cells of the blood vessels in pseudogout specimens. Intra- as well as extracellular crystals could also be demonstrated with the aid of scanning electron microscopy in sediments of synovial fluid in gout and pseudogout. Transmission electron microscopical investigations of synovial tissue specimens detected proliferated and pleomorphic synovial lining cells in gout in contrast to a more monomorphic appearance of these cells in pseudogout. The crystals were washed out during the preparation techniques for transmission electron microscopy so that needle-like empty spaces resulted within cytoplasm of the phagocytic cells. These clefts were surrounded by phagosomal structures and densified cytoplasmic ground substance; sometimes they were also lined by membranes...     

Expression of sialyl-Lewis X, an E-selectin ligand, in inflammation, immune processes, and lymphoid tissues.

The carbohydrate structure sialyl-Lewis X (SLex) can function as a ligand for E-selectin, formerly known as endothelial leukocyte adhesion molecule-1 (ELAM-1). This study was performed to analyze the expression of SLex by leukocytes and other cell types in the context of inflammatory and immune processes. Human peripheral blood cells were examined by flow cytometry using monoclonal antibody CSLEX1 directed against SLex. Cell surface SLex was found in abundance on nearly all isolated polymorphonuclear leukocytes (PMN) and monocytes, and at low levels on a substantial portion (up to 40%) of natural killer cells. This moiety was expressed also on approximately 10% of peripheral blood T cells. Immunohistochemistry was performed on various human tissues involved in inflammatory or immune processes and on secondary lymphoid tissues. In acute appendicitis, endothelial cells of postcapillary venules expressed E-selectin, and most PMN, both within vessels and extravasated, expressed SLex. A substantial number of monocytes/macrophages in inflamed appendiceal, synovial, and dermal tissues also reacted with antibody CSLEX1; however, only rare tissue macrophages in uninflamed nonlymphoid sites showed expression of SLex. These observations are consistent with the concept that SLex on circulating PMN and monocytes functions as a ligand for endothelial E-selectin in the development of inflammatory reactions. SLex-positive lymphocytes also were seen, notably, T lymphocytes in inflamed skin. An unexpected finding was that the CSLEX1 antibody also reacted with venular endothelium in certain lymphoid tissues and in inflamed appendix, but not with endothelium in normal appendix. Whether the SLex antigen identified on endothelium represents de novo expression or passive adsorption remains to be determined.     

In vivo role of phagocytic synovial lining cells in onset of experimental arthritis.

The in vivo role of phagocytic synovial lining cells (SLC) was studied in acute experimental arthritis in the mouse. SLCs were selectively depleted by injecting liposomes encapsulating the drug dichloromethylene diphosphonate (CL2MDP, Clodronate). Optimal depletion of phagocytic lining cells occurred 7 days after CL2MDP liposome injection. Eliciting an immune complex-mediated arthritis in SLC-depleted knee joints largely prevented inflammation if compared to control arthritic knee joints. Joint swelling and influx of inflammatory cells into the joint cavity was markedly diminished. Cartilage damage, in this model related to influx of inflammatory cells, was significantly decreased. Reduced influx of inflammatory cells (mainly polymorphonuclear neutrophils) was correlated to a decreased production of chemotactic factors as measured in washouts of arthritic joints in a two-compartment Transwell system. Interleukin-1-driven chemotactic factors seem to be involved. Interleukin-1 levels were significantly lowered in SLC-depleted arthritic knee joints as compared to controls. Injection of recombinant murine interleukin-1 in SLC-depleted knee joints caused less influx of inflammatory cells as compared to injection into control knee joints. A specific damage of CL2MDP liposome treatment to synovial blood vessels was excluded as intraarticular injection of human recombinant C5a in lining-depleted knee joints showed similar influx of inflammatory cells if compared to human recombinant C5a injection in control knee joints. This study indicates that in immune complex-mediated arthritis, phagocytic lining cells regulate the onset of the inflammatory response.     

Immunohistochemical analysis of synovial membranes from inflammatory and non-inflammatory arthritides: scarcity of CD5 positive B cells and IL2 receptor bearing T cells.

Rheumatoid Arthritis (RA) synovial membranes were examined by single and dual immunohistological techniques with a number of monoclonal antibodies against lymphocyte and macrophage related antigens. CD4 positive T lymphocytes frequently expressed MHC Class II antigens and were found in sublining collections in close association with activated macrophages as well as B lymphocytes. CD8 positive T cells surrounded these collections as well as being scattered throughout the membrane and also frequently expressed MHC Class II antigens. IL2 receptor (IL2r) expression on T cells and CD5 expression on B cells were rarely seen in these synovial membranes. Similar immunohistological architecture was found in synovial membranes from patients with psoriatic arthritis (PA) and Reiter's Syndrome (RS). Normal synovium contained few T cells, with few cells expressing MHC Class II antigens. Synovium from osteoarthritis (OA) patients also demonstrated similar immunohistological changes to those found in inflammatory arthritides, suggesting that there are only quantitative rather than qualitative differences between the synovial membrane immunohistological architecture from patients with inflammatory and noninflammatory arthritides.     

Gross and histopathological findings in synovial membranes of pigs with experimentally induced Mycoplasma hyosynoviae arthritis

The aim of this investigation was to describe the gross and histopathological findings in synovial membranes of pigs with experimentally induced Mycoplasma hyosynoviae arthritis and in synovial membranes of non-infected pigs of the same age. Experimental intravenous or intranasal inoculation or contact exposure with M. hyosynoviae induced arthritis in 13- to 17-week-old pigs. The acute to subacute arthritis was characterized by increased amounts of serohaemorrhagic, serofibrinous or mahogany coloured synovial fluid combined with edema and hyperaemia, followed by yellow to brownish discoloration and moderate villous proliferation of the synovial membrane. In the chronic phase moderate fibrosis was seen, but no periarticular or articular cartilage involvement. The acute to subacute histopathological characteristics were edema, hyperaemia, variable hyperplasia of synovial lining cells, increased density of subsynovial cell populations, diffuse and perivascular infiltration with lymphocytes, plasma cells and macrophage-like cells, fibrinous material, mild to moderate villous hypertrophy and mild to moderate fibrosis in chronic cases. The morphogenetic changes during the course of the infection may be described as follows: An acute preimmune phase with inflammatory changes and synovial membrane reactions dominates the first week of the infection. By the second and third week, the peak of the immune phase with masses of plasma cells and lymphocytes is seen. By 7 weeks, there is healing with moderate fibrosis. Mild ongoing reactions or recurrence of arthritis, perhaps related to persistence of mycoplasma antigens, may be seen in later phases and local antibody production may be important in this infection.     

The proinflammatory effect of intra-articular injection of soluble human and venom phospholipase A2.

The proinflammatory effects of intra-articular injection of purified phospholipase A2 from snake venom and rheumatoid synovial fluid were studied in rats. Purified soluble phospholipase A2 (PLA2) in concentrations ranging from 1000 to 20,000 units/ml, was injected intra-articularly. Histologic parameters examined were cell and protein content of synovial fluid, subsynovial cellular infiltration, synovial lining cell hyperplasia, bone erosion, and peri-articular soft tissue infiltration. Single intra-articular injections of PLA2 resulted in an acute inflammatory infiltrate of the subsynovium with maximal changes seen 2 to 6 hours after injection. Acute inflammatory changes were dose-dependent. Joints injected repeatedly at 24-hour intervals showed prominent synovial lining cell hyperplasia, maximal at 96 hours. Human synovial and snake venom PLA2s were equipotent at inducing both the acute and chronic articular changes. These changes were not seen in joints injected with inactivated PLA2. It is concluded that soluble PLA2 causes time- and dose-dependent acute inflammatory changes after a single intra-articular injection and synovial lining cell hyperplasia in response to repeated exposure to PLA2. The experimental proliferative synovitis in this model may correlate with features of acutely inflammed joints bathed in synovial fluids containing high levels of PLA2 in patients with rheumatoid arthritis.     

The production of arthritis in the guinea-pig by intra-articular reaction between lymphokines and inflammatory leucocytes.

A single intra-articular injection of lymphokine into the guinea-pig knee joint resulted in a sequence of changes in joint architecture whose histopathological features resembled that of an acute inflammatory reaction progressing to a chronic state. At 24 h there was a mild hyperplasia and hypertrophy of the synovium with intense polymorphonuclear leucocyte infiltration. At 72 h, the synovium was heavily infiltrated with diffuse and focal aggregations of mononuclear cells; erosion of cartilage and bone by synovial pannus was accompanied by a subsynovial fibrosis. By 1 week, leucocytic infiltration of the synovium had decreased markedly although the erosion and fibrosis persisted. However, when lymphokine was injected together with oil-elicited peritoneal exudate cells a more intense arthritis ensued: at 72 h synovial pannus was prominently eroding bone and this was accompanied by the appearance of multinucleate cells resembling osteoclasts in the zone of erosion. These features were shown to resemble closely the histopathology of experimental allergic arthritis in the guinea-pig, in contrast to the lesser severity of synovitis resulting from the adoptive cellular transfer of delayed hypersensitivity into the joint. The results indicate that lymphokines may play a role in the induction of experimental allergic arthritis by recruiting and activating cells involved in chronic inflammation.     

The production of arthritis in the guinea-pig by intra-articular reaction between lymphokines and inflammatory leucocytes

A single intra-articular injection of lymphokine into the guinea-pig knee joint resulted in a sequence of changes in joint architecture whose histopathological features resembled that of an acute inflammatory reaction progressing to a chronic state. At 24 h there was a mild hyperplasia and hypertrophy of the synovium with intense polymorphonuclear leucocyte infiltration. At 72 h, the synovium was heavily infiltrated with diffuse and focal aggregations of mononuclear cells; erosion of cartilage and bone by synovial pannus was accompanied by a subsynovial fibrosis. By 1 week, leucocytic infiltration of the synovium had decreased markedly although the erosion and fibrosis persisted. However, when lymphokine was injected together with oil-elicited peritoneal exudate cells a more intense arthritis ensued: at 72 h synovial pannus was prominently eroding bone and this was accompanied by the appearance of multinucleate cells resembling osteoclasts in the zone of erosion. These features were shown to resemble closely the histopathology of experimental allergic arthritis in the guinea-pig, in contrast to the lesser severity of synovitis resulting from the adoptive cellular transfer of delayed hypersensitivity into the joint. The results indicate that lymphokines may play a role in the induction of experimental allergic arthritis by recruiting and activating cells involved in chronic inflammation.     

Experimental immune inflammation in the synovial membrane: I. The immunological mechanism

Synovitis was produced in immunized guinea-pigs and rats by injection of antigen into the knee joint. The reaction was mainly mononuclear at 48 hours and, in the guinea-pig, progressed to chronic granuloma formation. Antigens principally used were bovine γ-globulin and tuberculin PPD. Immune deviation, giving a diminished delayed hypersensitivity response, also gave diminished synovial inflammation when compared with undeviated control animals. Immune synovitis to tuberculin PPD was successfully transferred with peritoneal cells taken from immunized animals and given intravenously to normal recipients whose knee joints were injected with antigen. Intravenous transfer of immune serum gave rise to a synovial reaction to bovine γ-globulin, injected into recipients' joints. Transfer of both cells and serum gave rise to particularly severe reactions. Transfer of either cells or serum or cells with serum failed to give reactions extending beyond 3–4 days. This suggests that active immunization is a requisite for the chronic inflammatory reaction of synovitis. Although the 48-hour synovial membrane reaction of rats was similar to that seen in the guinea-pig, chronic inflammatory reactions were not found in that species.     

膝关节痛风性关节炎的超声诊断价值

目的 探讨超声在痛风性关节炎诊断中的价值。方法 选取2015年3月~2018年12月我院确诊为痛风性关节炎且膝关节受累患者20例,均进行超声检查及关节镜检查,观察患者超声及关节镜下表现。结果 超声检查:声像图异常表现有滑膜增生、积液、尿酸结晶沉积、痛风石形成、“双轨征”、骨侵蚀,其中可见滑膜增生19例、积液15例、尿酸结晶沉积13例、痛风石形成5例、“双轨征”3例、骨侵蚀1例。关节镜检查:镜下可见大量白色尿酸盐结晶,沉积于关节腔内。结论 超声检查能有效、全面的评估痛风性关节炎患者受累关节情况,能够实时动态对不同病程阶段的关节情况进行监测,其可作为痛风性关节诊断的重要影像学手段。     

Supravital staining of synovial fluid with Testsimplets.

We explored the use of Testsimplet (TS) in synovial fluid (SF) analysis. TS is a glass slide coated with a dry mixture of methylene blue and cresyl violet, which in contact with one drop of SF provides a stained fresh preparation. We applied the TS to the study of 159 SFs of patients with different rheumatic diseases. In those SFs of patients with crystal-associated diseases, the crystal search was performed both on unstained preparations and with TS. TS was as good as the Wright's and Papanicolaou stain in characterizing SF cells, lupus erythematosus cells, and detection of occasional bacteria. TS allowed a better visualization of Reiter's cells, cartilage fragments, synovial villi, fat droplets, and fibrin. Crystals were identified in every TS of those patients with crystal-associated diseases. TS is a rapid and reproducible method of SF supravital staining. Crystals are well preserved for simultaneous examination with compensated polarized light.     

ALVEOLAR DESTRUCTION IN EXPERIMENTAL KLEBSIELLA PNEUMONIA

Sequential histological changes of the lungs were studied in experimental Klebsiella pneumonia, using untreated control mice, cyclophosphamide-treated mice, and carrageenan-treated mice. Cyclophosphamide was used to deplete polymorphonuclear leukocytes and monocytes, and carrageenan was used to deplete mononuclear phagocytes selectively. At 72 hours, varying degree of alveolar necrosis could be seen in untreated control mice. However, the lung lesions of cyclophosphamide- or carrageenan-treated mice were significantly different from those of the control mice. The lung lesions of cyclophosphamide-treated mice indicated that destruction of the alveolar septa was not induced by K. pneumoniae itself but by inflammatory cells, because the alveolar walls were preserved very well in spite of considerable bacterial multiplication in alveolar lumina until infiltration of inflammatory cells occurred. The lung lesions of carrageenan-treated mice showed that alveolar spaces were packed with polymorphonuclear leukocytes, but the alveolar walls were preserved very well as far as the authors could tell after examining the lung lesions by silver impregnation staining. These results suggest that macrophages rather than polymorphonuclear leukocytes and organisms play an important role in alveolar injury in experimental Klebsiella pneumonia.     

Alveolar destruction in experimental Klebsiella pneumonia

Sequential histological changes of the lungs were studied in experimental Klebsiella pneumonia, using untreated control mice, cyclophosphamide-treated mice, and carrageenan-treated mice. Cyclophosphamide was used to deplete polymorphonuclear leukocytes and monocytes, and carrageenan was used to deplete mononuclear phagocytes selectively. At 72 hours, varying degree of alveolar necrosis could be seen in untreated control mice. However, the lung lesions of cyclophosphamide- or carrageenan-treated mice were significantly different from those of the control mice. The lung lesions of cyclophosphamide-treated mice indicated that destruction of the alveolar septa was not induced by K. pneumoniae itself but by inflammatory cells, because the alveolar walls were preserved very well in spite of considerable bacterial multiplication in alveolar lumina until infiltration of inflammatory cells occurred. The lung lesions of carrageenan-treated mice showed that alveolar spaces were packed with polymorphonuclear leukocytes, but the alveolar walls were preserved very well as far as the authors could tell after examining the lung lesions by silver impregnation staining. These results suggest that macrophages rather than polymorphonuclear leukocytes and organisms play an important role in alveolar injury in experimental Klebsiella pneumonia.     

PARTICIPATION OF MONOCYTES IN GLOMERULONEPHRITIS IN ACUTE SERUM SICKNESS OF RABBIT

Acute serum sickness In the rabbit was studied with special reference to the role of monocytes in the Inflammatory process in the glomerulus. It was revealed that macrophages were the major factor in producing glomerular hyperceliularity In acute serum sickness. Proliferation of intrinsic glomerular cells or accumulation of polymorphonuclear leukocytes (PMNs) was minimal. Ultrastructural characteristics of these phagocytic cells were described. Macrophages engulfed various inflammatory products such as fibrin and cell debri in the glomerular capillary. Colloidal carbon administered at the active inflammatory stage was found to be mostly engulfed by macrophages, little by mesangial cells, and was not seen In endothelial or epithelial cells and PMNs. The selective ingestion of the carbon particles by these macrophages made it possible to differentiate them from glomerular cells. This in turn indicated that the macrophages were derived from neither endothelial nor mesangial cells and that they were of blood monocytic origin. It was suggested that monocytic cells participated in glomerular Inflammation but they, on the other hand, contributed to the repair of glomerular Injuries through their active role for phagocytosis.