Anti-phospholipid (aPL) antibodies that exhibit binding in cardiolipin (CL) ELISA can be purified to >95% purity by sequential phospholipid affinity and ionexchange chromatography. However, these highly purified aPL antibodies do not bind to the CL antigen when assayed by a modified CL ELISA in which the blocking' agent does not contain bovine serum, nor do they bind to phospholipid affinity columns. Binding to the phospholipid antigen will Anti-phospholipid (aPL) antibodies are autoantibodies that can be detected in plasma or serum in solid-phase immunoassays in which negatively charged phospholipids, most commonly cardiolipin (CL), are used as the antigen (1, 2). We have previously described a simple two-step procedure for purifying aPL antibodies from plasma (or serum) by sequential phospholipid affinity and cation-exchange chromatography, yielding specific immunoglobulin of >95% purity (3). These antibodies exhibit typical binding in CL ELISA but do not possess lupus anticoagulant (LA) activity in phospholipid-dependent clotting tests. Recently, we have also shown that plasma can be resolved by ion-exchange chromatography into fractions containing either anticardiolipin (aCL) antibodies or antibodies with LA activity, strongly suggesting that aCL and LA antibodies represent distinct antibody subgroups (4). Although antibodies binding CL in immunoassays also generally bind all anionic phospholipids (2, 4), and hence are best referred to as aPL antibodies, we hereafter refer to this group as aCL antibodies in distinction to LA, which may also be aPL antibodies but appear to be directed against a different antigen (4).We have previously noted that when aCL antibodycontaining fractions derived from ion-exchange chromatography of plasma were applied to phosphatidylserine or CL affinity columns, there was no binding ofthe antibody despite the fact that when plasma containing these antibodies was applied to these columns, aCL antibodies could be purified. This suggested that there was a cofactor also present in plasma or serum that was required for aCL antibodies to bind to the affinity columns. Addition of normal (aCL antibody negative) plasma to the ion-exchange fractions resulted in aCL binding to the columns supporting this hypothesis (4).In this report, we have further investigated this phenomenon. We have found that the plasma cofactor is also required for aCL antibodies to bind CL in a modified immunoassay in which bovine serum (which also contains the cofactor) is excluded. We have been able to purify the cofactor to homogeneity and identify it as /2-glycoprotein I (p32GPI), also known as apolipoprotein H. These results may change our understanding of the biology of aCL antibodies. MATERIALS AND METHODSPlasma and Patients. Citrated platelet-free plasma was prepared by adding freshly drawn blood from venipuncture into tubes containing 1/10th final vol of 0.11 M trisodium citrate, immediate centrifugation at 2500 x g for 15 min, and filtration through a 0.22-pum Millipore Millex filter (4). aCL anti...

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Objective. Systemic lupus erythematosus (SLE) is a chronic inflammatory autoimmune disease that significantly impairs patients' quality of life and can be life threatening. This study aimed to describe the experiences and perspectives of adults living with SLE. Methods. We conducted a systematic review and thematic synthesis of qualitative studies that explored the experiences of adults living with SLE. We searched MEDLINE, Embase, PsycINFO, CINAHL (to November week 1, 2012), Google Scholar, a thesis database, and reference lists of relevant articles. Results. Forty-six studies involving 1,385 participants were included. Five themes were identified: restricted lifestyle (including subthemes of pervasive pain, debilitating fatigue, mental deterioration, disruptive episodic symptoms, and postponing parenthood), disrupted identity (gaining diagnostic closure, prognostic uncertainty, being a burden, hopelessness, heightened self-consciousness, fearing rejection, and guilt and punishment), societal stigma and indifference (illness trivialization, socially ostracized, and averse to differential treatment), gaining resilience (optimism, control and empowerment, being informed and involved, and valuing mutual understanding), and treatment adherence (preserving health, rapport with clinicians, negotiating medication regimens, and financial burden). Conclusion. SLE has a severe and pervasive impact on patients' self-esteem and independence. Their physical and social functioning is limited and they feel anxious about their future. Patients perceive that SLE is trivialized, misunderstood, and stigmatized by their family, friends, and physicians, which intensifies their sense of isolation. Educational, psychosocial, and self-care interventions are needed to promote mental resilience, positive coping strategies, self-advocacy, and capacities for social participation, and thereby to achieve better treatment and health outcomes in patients with SLE. INTRODUCTIONSystemic lupus erythematosus (SLE) is a chronic autoimmune disease with a relapsing and remitting course that can be life threatening. The disease has a peak incidence in females ages 15-40 years, and the prevalence of SLE is higher in nonwhite populations (1-4). Although treatment advances have reduced mortality rates in SLE, patients still experience impaired quality of life (QOL) and long-term morbidity (3,5-7).The debilitating pain, musculoskeletal manifestations, fatigue, and renal and cutaneous problems can limit patients' ability to work and participate in family and social activities (8 -10). The psychological and cognitive impact of SLE can have an adverse effect on physical functioning and disease activity (10 -12). Depression may be associated with greater physical disability, and low self-esteem has been found to be associated with greater cumulative organ damage (13). Moreover, studies have identified discordances between patients and clinicians in their perceptions of disease burden and activity, and consequently this can lead to poor treatment adherence, d...

C-reactive protein (CRP) and serum amyloid A (SAA) increase in the blood of patients with inflammatory conditions and CRP-induced monocyte tissue factor (TF) may contribute to inflammation-associated thrombosis. This study demonstrates that SAA is a potent and rapid inducer of human monocyte TF. SAA induced TF mRNA in PBMC within 30 min and optimal procoagulant activity within 4 h, whereas CRP (25 μg/ml)-induced activity was minimal at this time. Unlike CRP, SAA did not synergize with LPS. Procoagulant activity was inhibited by anti-TF and was dependent on factors VII and X, and TF Ag levels were elevated on CD14+ monocytes. Responses were optimal with lymphocytes, although these were not obligatory. Inhibitor studies indicate activation of NF-κB through the ERK1/2 and p38 MAPK pathways; the cyclo-oxygenase pathway was not involved. SAA-induced TF was partially inhibited by high-density lipoprotein, but not by low-density lipoprotein or by apolipoprotein A-I. SAA is a ligand for the receptor for advanced glycation end products (RAGE), and TF generation was suppressed by ∼50% by a RAGE competitor, soluble RAGE, and by ∼85% by anti-RAGE IgG. However, another RAGE ligand, high mobility group box-1 protein, capable of inducing monocyte chemotactic protein-1 mRNA in 2 h, did not induce TF within 24 h. Cross-linking studies confirmed SAA binding to soluble RAGE. Elevated SAA is a marker of disease activity in patients with rheumatoid arthritis, and PBMC from patients with rheumatoid arthritis were more sensitive to SAA than normals, suggesting a new link between inflammation and thrombosis.

Anticardiolipin antibodies (aCL) purified from patients with autoimmune disease have recently been shown to interact with a phospholipid-binding plasma protein, beta 2-glycoprotein I (beta 2-GPI). The aim of this study was to determine whether aCL purified from patients with infection also interact with beta 2-GPI. aCL purified from 23 patients with malaria, infectious mononucleosis, tuberculosis, hepatitis A or syphilis did not require the presence of beta 2-GPI to bind cardiolipin (CL). In contrast, aCL were purified from 11 out of 12 patients with autoimmune disease that bound CL only in the presence of beta 2-GPI. Thrombotic complications appear to be associated with aCL occurring in autoimmune disease but not with aCL associated with infections. We postulate that this increased risk of thrombosis in the autoimmune group may be due to the presence of aCL that bind CL in association with beta 2-GPI, a plasma protein with anticoagulant activity.

Although mast cells (MCs) often are abundant in the synovial tissues of patients with rheumatoid arthritis, the contribution of MCs to joint inflammation and cartilage loss remains poorly understood. MC-restricted tryptase/heparin complexes have proinflammatory activity, and significant amounts of human tryptase β (hTryptase-β) are present in rheumatoid arthritis synovial fluid. Mouse MC protease-6 (mMCP-6) is the ortholog of hTryptase-β, and this serine protease is abundant in the synovium of arthritic mice. We now report that C57BL/6 (B6) mice lacking their tryptase/heparin complexes have attenuated arthritic responses, with mMCP-6 as the dominant tryptase responsible for augmenting neutrophil infiltration in the K/BxN mouse serum-transfer arthritis model. While inflammation in this experimental arthritis model was not dependent on protease-activated receptor-2, it was dependent on the chemokine receptor CXCR2. In support of the latter data, exposure of synovial fibroblasts to hTryptase-β/heparin or mMCP-6/heparin complexes resulted in expression of the neutrophil chemotactic factors CXCL1/KC, CXCL5/LIX, and CXCL8/IL-8. Our proteomics, histochemistry, and immunohistochemistry data also revealed substantial loss of cartilage-derived aggrecan proteoglycans in the arthritic joints of wild-type B6 mice but not mMCP-6-null B6 mice. These observations demonstrate the functional contribution of MC-restricted tryptase/heparin complexes in the K/BxN mouse arthritis model and connect our mouse findings with rheumatoid arthritis pathophysiology.

Calgranulins comprise three proteins, S100A8 (Calgranulin A), S100A9 (Calgranulin B) and S100A12 (Calgranulin C) that are predominantly expressed by neutrophils, monocytes and activated macrophages. These S100 calcium-binding proteins are important molecular mediators in a range of diseases, including inflammatory arthritis, atherosclerosis and microbial infections. Much of the literature has focused on the pro-inflammatory functions of calgranulins, and this has tended to underplay important regulatory, anti-oxidant and protective properties. S100A8 and S100A9 are particularly complex in their actions because they exert intracellular and extracellular functions, they form a heterocomplex, S100A8-A9 (calprotectin), and have actions that are independent of or dependent on heterocomplex formation. In some circumstances S100A9 appears to regulate, rather than synergize with the actions of S100A8 and vice versa. Moreover, these calgranulins also bind zinc and other divalent cations and are sensitive to post-translational oxidative modifications, properties that also affect some functions. It is important to note that S100A8 has potent anti-oxidant activity, which could be important in host protection. Furthermore, although the genes for S100A8 and S100A9 are induced by activation of the toll-like receptor/interleukin-1 pathway, their expression is enhanced by interleukin-10 and glucocorticoids, thus suggesting a regulatory role in inflammation. On the other hand, S100A12 appears to be predominantly pro-inflammatory, particularly by its ability to activate mast cells. Measurement of S100A12 levels may be a highly sensitive biomarker for inflammatory disease activity. This review summarizes the current understanding of the biology of calgranulins, with a focus on their pleiotropic roles in inflammatory arthritis. Keywords: calgranulin; inflammation; S100A8; S100A9; S100A12; arthritisThe inflammatory arthritides comprise a group of conditions in which the primary pathology involves acute or chronic inflammation of synovium, the specialized tissue lining diarthrodial joint cavities. If untreated, inflamed synovium (synovitis) mediates progressive damage and erosion to the key tissues that enable efficient joint function: articular cartilage, muscle tendons, periarticular ligaments and subchondral bone. These two key processes, synovial inflammation and joint tissue damage contribute to the clinical symptoms of pain, swelling, stiffness, loss of function and impairment of health suffered by patients with acute and chronic inflammatory arthritides. 1 Important features of chronic synovitis include hyperplasia of the synovial lining layer cells, infiltration of abundant numbers of leucocytes and formation of new blood vessels (angiogenesis). Virtually all cells involved in inflammation are expanded or recruited to the inflamed synovium, including resident fibroblast-like and macrophage-like synoviocytes, mast cells, neutrophils which migrate through synovium to the synovial fluid space, monocyte/macrophages, dendritic cell...