The semi-quantitative scoring system recommended here is simple to apply and required no specialized equipment. Scoring of the tibial plateaus was highly reproducible and more consistent than that of the femur due to the much thinner femoral cartilage. This scoring system may be a useful tool for both new and experienced scorers to sensitively evaluate models and OA mechanisms, and also provide a common paradigm for comparative evaluation across the many groups performing these analyses.

Objective. Modification of antigens represents a trigger for the generation of autoantibodies. In the pathogenesis of rheumatoid arthritis (RA), citrullination of proteins has been shown to be a critical process, and the determination of antibodies against citrullinated antigens has been a diagnostic milestone. We undertook this study to determine whether antibodies to mutated and citrullinated vimentin (MCV) could serve as a diagnostic and prognostic marker for RA.Methods. We identified novel isoforms of human MCV in the synovial fluid of RA patients. The significance of these disease-related modifications was investigated by the analysis of autoantibody reactivities. In a group of 1,151 RA patients, the diagnostic significance and the prognostic value of an anti-MCV enzyme-linked immunosorbent assay (ELISA) were compared with that of an anti-cyclic citrullinated peptide (anti-CCP) ELISA.Results. In RA, sensitivities of 82% and 72% were calculated for the anti-MCV and anti-CCP assays, respectively. The specificity of both assays was comparable (98% and 96%, respectively). In followup analyses of 16 RA patients with moderate disease activity (mean Disease Activity Score in 28 joints [DAS28] of 2.72) and 26 RA patients with active disease (mean DAS28 of 5.07), disease stratification of RA was possible using the anti-MCV assay (P ‫؍‬ 0.0084). A significant correlation of anti-MCV antibodies with the DAS28 was documented (r ‫؍‬ 0.5334, P ‫؍‬ 0.0003), in 42 RA patients (n ‫؍‬ 427 antibody determinations at different time points).

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We studied the presence of bacterial antigens in rat tissues.We produced a monoclonal antibody (MAb 2E9) directed against intestinat flora-derived peptidoglycan-polpaharide complexes from human and rat feces. With several immunological techniques, the specificity of 2E9 for this bacterial product was demonstrated. Using 2E9 in an immunohistological assay, we were able to show the presence of bacterial products in macrophages in the red pulp of spleens of conventional Lewis rats. However, we found no correlation between the development of the intestinal flora and positive spleen staining with MAb 2E9. The results were confirmed

Insulin-like growth factor-1 (IGF-1) is an essential anabolic growth factor in the regulation of cartilage metabolism and exerts its effects by binding to the IGF-1 Type 1 receptor on the chondrocyte membrane. We have localized and quantified in situ IGF-1 receptor expression in intact articular cartilage of normal mice. The IGF-1 receptor was detected immunohistochemically with antibodies to the IGF-1 receptor and visualized with conventional light microscopy and confocal laser scanning miaoscopy (CLSM). CLSM analysis enabled us to distinguish IGF-1 receptor immunoreactivity on the chondrocyte cell membrane from intracellular staining. We have established two approaches to quantitir in confocal images low levels of fluorescence intensity of the immuno-

Lower numbers of neuropeptide-containing fibers in arthritic joints have been found as compared to control joints. This may be the result of fiber depletion, necrosis of fibers, or proliferation of soft tissues without neural sprouting. To discriminate between these possibilities, we studied the relationships between soft tissue proliferation, changes in vascularity of synovial tissues, and changes in joint innervation during arthritis. Arthritis was induced in the knee joint of mice by a single subpatellar injection of methylated bovine serum albumin after previous immunization. Antibodies to protein gene product 9.5, S-100, and growth-associated protein-43 (GAP-43) were used to study the general innervation pattern. Antibodies to calcitonin gene-related peptide (CGRP), vasointestinal polypeptide (VIP), substance P (SP), and tyrosine hydroxylase (TH) were used to localize sensory (SP, CGRP, VIP) and sympathetic (TH) fibers. Blood vessels of the joint were studied with ink perfusion, GAP-43, and a vascular marker (LF1). Directly after the induction of arthritis, the synovial cavity was enlarged and filled with leukocytes. From day 4 onward, small sprouting blood vessels penetrated the avascular mass of cells in the joint cavity. After 1 week, the vascular sprouting activity and GAP-43 immunoreactivity were maximal, and after 2 weeks, vascular sprouting activity diminished. In the subsequent period, the synovia slowly regained their prearthritic appearance and thickness. The most pronounced changes in the general staining pattern of CGRP, SP, VIP, and TH were found in the periosteum. From 2 days to 4 weeks after the induction of arthritis, the layer of SP, CGRP, and VIP fibers in the femoral periosteum was thicker and more irregular. GAP-43 staining showed many terminal varicosities, which suggested sprouting of nerve fibers. From 2 days to 2 weeks after the induction of arthritis, the SP and CGRP fibers in the periosteum showed gradual depletion. In the thickened subsynovial tissues that were revascularized, no ingrowth of neural elements was found. As the total number of nerve fibers in the synovial tissue did not change, large parts of the synovia directly facing the joint cavity were not innervated at 1 week after the induction of arthritis. These results strongly suggest that periosteal SP and CGRP fibers were depleted during arthritis. Synovial proliferation without concomitant fiber growth is the main cause of the reduced number of immunocytochemically detectable fibers in the mouse arthritic knee joint.

SummaryAlpha-fodrin, an intracellular organ-specific cytoskeleton protein is a recently identified autoantigen associated with Sicca-and Sjögren's syndrome (SS). SS frequently affects patients with Graves' ophthalmopathy (GO). We have therefore cloned and expressed the human recombinant 120-kDa fodrin-fragment. A sequential purification procedure was applied to isolate the recombinant protein. Using sera from patients with SS, the antigenicity of the purified fodrin fragment was demonstrated by immunoblotting. Sera from 144 patients with GO and 1200 blood donors were screened for the presence of anti-a a a a -fodrin IgA and IgG antibodies by a newly developed ELISA using the human a a a a -fodrin fragment as an autoantigen. In contrast to controls (< < < < 1% IgA only, P < < < < 0·001) and to subjects with various autoimmune diseases ( P < < < < 0·001), a a a a -fodrin antibodies were detected in 22% of patients with GO ( n = = = = 32). IgA and IgG antibodies were present in 21 (15%) and 14 (10%) GO subjects, respectively. A total of 45 patients with GO (31%) had at least one fodrin-or SS-antibody. GO patients with SS showed SS-and high titres of a a a a -fodrin-antibodies. In GO patients, fodrin antibodies correlated with TPO-( P < < < < 0·05) and SS-A ( P = = = = 0·002) antibodies. Thus, for the first time, antibodies reactive with fodrin are reported in patients with GO.

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