Targeted delivery of anti-inflammatory therapy to rheumatoid tissue by fusion proteins containing an IL-4-linked synovial targeting peptide

We provide first-time evidence that the synovial endothelium-targeting peptide (SyETP) CKSTHDRLC successfully delivers conjugated IL-4 to human rheumatoid synovium transplanted into SCID mice. SyETP, previously isolated by in vivo phage display and shown to preferentially localize to synovial xenografts, was linked by recombinant technology to hIL-4 via an MMP-cleavable sequence. Both IL-4 and the MMP-cleavable sequence were shown to be functional. IL-4-SyETP augmented production of IL-1ra by synoviocytes stimulated with IL-1[beta] in a dose-dependent manner. In vivo imaging confirmed increased retention of SyETP-linked-IL-4 in synovial grafts which was enhanced by increasing number of copies (one to three) in the constructs. Strikingly, SyETP delivered bioactive IL-4 in vivo as demonstrated by increased pSTAT6 in synovial grafts. Thus, this study provides proof of concept for peptide-tissue-specific targeted immunotherapy in rheumatoid arthritis. This technology is potentially applicable to other biological therapies providing enhanced potency to inflammatory sites and reducing systemic toxicity

Expression and Activities of Matrix Metalloproteinases under Oscillatory Shear in IL-1-Stimulated Synovial Cells

Matrix metalloproteinases (MMPs) are known to play a critical role in tissue disintegration, and an elevated level of MMPs is observed in synovium and synovial fluid of joints with rheumatoid arthritis. During joint movement, synovial tissue receives various mechanical stimuli, but effects of mechanical challenges on regulation of MMPs in rheumatic synovium are poorly understood. Focusing on cellular responses to oscillatory fluid shear in human synovial cells, we determined the expression of MMP-1 and MMP-13 by polymerase chain reaction and immunoblotting as well as proteolytic activities of total MMPs by a fibril degradation assay and zymography. The results revealed that ~0.5 dyn/cm 2 oscillatory shear at 1 Hz not only reduced an mRNA level and a protein level of MMP-1 and MMP-13, but it also decreased collagenase and gelatinase activities of total MMPs. Furthermore, the induction of the MMP expression and activities by interleukin-1 was suppressed by the oscillatory shear. Interestingly, the oscillatory shear upregulated the mRNA expression of TIMP-1 and TIMP-2. Our results support a potential role of oscillatory shear in regulating expression and activities of MMPs in the presence and the absence of proinflammatory cytokine

Analysis of the temporal expression of chemokines and chemokine receptors during experimental granulomatous inflammation: role and expression of MIP-1Α and MCP-1

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/72640/1/sj.bjp.0704371.pd

The potential role of kallistatin in the development of Abdominal Aortic Aneurysm

Abdominal aortic aneurysm (AAA) is a vascular condition that causes permanent dilation of the abdominal aorta, which can lead to death due to aortic rupture. The only treatment for AAA is surgical repair, and there is no current drug treatment for AAA. Aortic inflammation, vascular smooth muscle cell apoptosis, angiogenesis, oxidative stress and vascular remodeling are implicated in AAA pathogenesis. Kallistatin is a serine proteinase inhibitor, which has been shown to have a variety of functions, potentially relevant in AAA pathogenesis. Kallistatin has been reported to have inhibitory effects on tumor necrosis factor alpha (TNF-α) signaling induced oxidative stress and apoptosis. Kallistatin also inhibits vascular endothelial growth factor (VEGF) and Wnt canonical signaling, which promote inflammation, angiogenesis, and vascular remodeling in various pre-clinical experimental models. This review explores the potential protective role of kallistatin in AAA pathogenesis

Role of C‐X‐C chemokines as regulators of angiogenesis in lung cancer

Lung cancer is the leading cause of malignancy‐related mortality in the U.S. and is predicted to increase over the remainder of this decade. Despite attempts to advance early diagnosis and use combination therapies, the clinical response of this cancer yields an overall 5‐year survival rate of less than 15%. Clearly, new strategies for therapy are indicated. Although carcinogenesis is complex, tumor growth beyond 1–2 mm3 is dependent on angiogenesis. One of the potential mechanisms that allows for tumorigenesis is dysregulation of the balance of angiogenic and angiostatic factors that favors net neovascularization within the primary tumor. Numerous studies have investigated the role of a variety of molecules in the regulation of angiogenesis. Recently, interleukin‐8 (IL‐8), a member of the C‐X‐C chemokine family, has been found to be an angiogenic factor. In contrast, platelet factor 4 (PF4), another C‐X‐C chemokine, has been shown to have angiostatic properties. It is interesting that the major structural difference between IL‐8 and PF4 is the presence of the NH2‐terminal ELR (Glu‐Leu‐Arg) motif that precedes the first cysteine amino acid residue of IL‐8 and is important in ligand/receptor interactions. We hypothesize that angiogenesis associated with tumorigenesis is dependent on members of the C‐X‐C chemokine family acting as either angiogenic or angiostatic factors. This paradigm predicts that the biological balance in the expression of these C‐X‐C chemokines dictates whether the neoplasm grows and develops metastatic potential or regresses. In this review we discuss our recent laboratory findings that support this contention and suggest that further elucidation of the biology of C‐X‐C chemokines in the context of neovascularization of nonsmall cell lung cancer will permit novel targeted therapy aimed specifically at attenuating tumor growth and metastasis. J. Leukoc. Biol. 57: 752–762; 1995.Peer Reviewedhttps://deepblue.lib.umich.edu/bitstream/2027.42/141888/1/jlb0752.pd

eNAMPT Is Localised to Areas of Cartilage Damage in Patients with Hip Osteoarthritis and Promotes Cartilage Catabolism and Inflammation

From MDPI via Jisc Publications RouterHistory: accepted 2021-06-21, pub-electronic 2021-06-23Publication status: PublishedFunder: Versus Arthritis; Grant(s): 21530,21812Funder: Medical Research Council; Grant(s): MR/K00414X/1Obesity increases the risk of hip osteoarthritis (OA). Recent studies have shown that adipokine extracellular nicotinamide phosphoribosyltransferase (eNAMPT or visfatin) induces the production of IL-6 and matrix metalloproteases (MMPs) in chondrocytes, suggesting it may promote articular cartilage degradation. However, neither the functional effects of extracellular visfatin on human articular cartilage tissue, nor its expression in the joint of hip OA patients of varying BMI, have been reported. Hip OA joint tissues were collected from patients undergoing joint replacement surgery. Cartilage explants were stimulated with recombinant human visfatin. Pro-inflammatory cytokines and MMPs were measured by ELISA and Luminex. Localisation of visfatin expression in cartilage tissue was determined by immunohistochemistry. Cartilage matrix degradation was determined by quantifying proteoglycan release. Expression of visfatin was elevated in the synovial tissue of hip OA patients who were obese, and was co-localised with MMP-13 in areas of cartilage damage. Visfatin promoted the degradation of hip OA cartilage proteoglycan and induced the production of pro-inflammatory cytokines (IL-6, MCP-1, CCL20, and CCL4) and MMPs. The elevated expression of visfatin in the obese hip OA joint, and its functional effects on hip cartilage tissue, suggests it plays a central role in the loss of cartilage integrity in obese patients with hip OA

Angiogenesis in interstitial lung diseases

Choroby śródmiąższowe płuc to bardzo duża grupa schorzeń o różnej etiologii i patogenezie. Dominującymi zaburzeniami w tych chorobach są przewlekłe zapalenie i włóknienie płuc. Procesy te, upośledzając dyfuzję gazów w płucach, prowadzą do hipoksji. Zarówno przewlekłe zapalenie, jak i hipoksja są bardzo silnymi bodźcami do nowotworzenia naczyń krwionośnych. Szczególnie ważną rolę odgrywa angiogeneza w chorobach nowotworowych, gdyż przyczynia się do wzrostu guza i tworzenia przerzutów. Zaobserwowano związki chorób śródmiąższowych płuc z nowotworami. Angiogeneza bierze również udział w procesie gojenia ran, umożliwiając napływ komórek do uszkodzonych struktur. Sugeruje się, że włóknienie płuc jest spowodowane zaburzonym procesem gojenia się ran. Wszystkie te obserwacje wskazują na udział angiogenezy w patogenezie śródmiąższowych chorób płuc. Udowodniono, że nowotworzenie naczyń krwionośnych odgrywa istotną rolę w rozwoju samoistnego włóknienia płuc. Coraz więcej danych wskazuje na udział neowaskularyzacji także w innych chorobach śródmiąższowych płuc, zarówno ziarniniakowych, jak i przebiegających z włóknieniem płuc lub z zapaleniem naczyń krwionośnych. Mechanizm tego procesu nie jest jeszcze w pełni wyjaśniony. Potwierdzenie udziału nowotworzenia naczyń krwionośnych w patogenezie chorób śródmiąższowych płuc stworzy możliwości zastosowania w leczeniu tych schorzeń nowych leków hamujących angiogenezę. W wybranych przypadkach w leczeniu tych chorób wykorzystywano już z powodzeniem inhibitory angiogenezy (np. talidomid).Interstitial lung diseases (ILD) constitute a large group of disorders characterized by various etiology and pathogenesis. Inflammation and pulmonary fibrosis are the most important processes in the course of ILD. Disease causes the decrease of the gas diffusion in the lungs and provokes hypoxia. Chronic inflammation and hypoxia are strong stimulus of neovascularization. Neoangiogenesis is a principal response of vessels to inflammation. The critical importance of tumor angiogenesis in the development and metastatic spread of tumors is proved. Relations of ILD with neoplasma have been observed. Neovascularization takes an important role in wound healing allowing the cells to flow into damaged structures. Recently, pulmonary fibrosis has been deemed to result from abnormal wound healing in the lung in response to injury to the alveolar epithelium. Angiogenesis participates in pathogenesis of idiopathic pulmonary fibrosis. More and more data suggest the role of angiogenesis in pathogenesis of other ILDs, such as granulomatosis, fibrosis and vasculitis. The mechanism of angiogenesis in ILD is not clear yet. New data concerning participation of neoangiogenesis in pathogenesis of ILD created target for new drugs. Thalidomide, a strong antiangiogenic drug was used successfully in the some cases of ILD

Estudo do lavado broncoalveolar de pacientes com paracoccidioidomicose pulmonar

Orientador: Maria Heloisa Souza Lima BlottaDissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciências MédicasResumo: Para investigar a resposta imune local versus sistêmica em pacientes com paracoccidioidomicose pulmonar analisamos o fenótipo de diferentes populações celulares e o padrão de produção de citocinas por células do lavado broncoalveolar (LBA) e do sangue periférico. O grupo estudado foi constituído por 19 pacientes com idade entre 36 a 65 anos. O diagnóstico foi confirmado pela demonstração do fungo P. brasiliensis no escarro, LBA e/ou biópsia. A análise do LBA mostrou aumento de linfócitos e neutrófilos em relação ao descrito para indivíduos normais saudáveis. Linfócitos T CD8+ encontravam-se em maior número no LBA do que no sangue periférico. A expressão de moléculas do MHC classe II, ICAM-1 (CD54) e B7-2, foi aproximadamente 3 vezes maior em macrófagos alveolares (MA), do que em monócitos do sangue periférico. Sobrenadantes de culturas de curta duração de MA apresentaram níveis mais elevados de IL-6, TNF-a, IL-12 p40 e MIP-1a, do que sobrenadantes de monócitos do sangue periférico. No LBA destes pacientes também foi possível detectar IL-6, TNF-a e MIP-1a, assim como anticorpos específicos anti-gp43 de P. brasiliensis, principalmente da classe IgG2. Como foi demonstrado que MIP-1a atrai seletivamente linfócitos T CD8+ e esta população celular está aumentada no LBA, nossos achados sugerem que além de macrófagos ativados, células T CD8+ devem ter um importante papel no desenvolvimento da PCM pulmonarAbstract: To investigate the local immune response we analyzed the cellular infiltrate and patterns of cytokine production in bronchoalveolar lavage cells and fluid from patients with pulmonar paracoccidioidomycosis (PCM). The group consisted of 19 patients, 16 male and 3 female, with age ranging from 36 to 65 years. The diagnosis was confirmed by demonstration of fungus in the sputum or bronchoalveolar lavage fluid (BAL), in addition to serological tests. Cytospin preparations from patients with PCM showed an increased number of lymphocytes in BAL, mostly CD8+T cells. Cultured alveolar macrophages produced higher levels of IL-6, TNF-a ??and MIP-1a as compared with PBMC. No differences were detected in relation to IL-8, IL-12p40, IL-10 and TGF-a. BAL fluid from PCM patients contained low but significant levels of IL-6, TNF-a and MIP-1a, and specific antibodies to Paracoccidioides brasiliensis, mainly of the IgG2 isotype. These findings indicate that the local inflammatory reaction in the lungs of patients with pulmonary paracoccidioidomycosis is mediated by the inflammatory cytokines TNF-a ??IL-6 and MIP-1a characterized by a lymphocytic infiltration and local release. MIP-1a may play an important role in the pathogenesis of PCM, mediating the recruitment of lymphocytes and macrophages to the site of infectionMestradoCiencias BiomedicasMestre em Ciências Médica

Cytokine and nitric oxide production in inflammatory arthritis

Rheumatoid arthritis (RA) is a chronic disease characterised by inflammatory infiltration of the synovial membrane, with concomitant destruction of adjacent cartilage and bone. Elucidation of immunoregulatory networks within the synovium offers the potential for therapeutic intervention. Two such pathways were investigated in the present study. Interleukin-15 (IL-15) is a novel pleiotropic cytokine produced by macrophages and fibroblasts, which induces T cell migration and activation and B cell maturation and immunoglobulin production. IL-15 was identified in RA synovial fluids and synovial membrane cultures and, using immunohistochemistry, its expression was localised in the RA synovial membrane to the lining layer and T lymphocyte aggregates. Enhanced proliferation and cytokine production to IL-15 was observed in RA synovial fluid (SF) T cells in comparison to matched peripheral blood (PB) T lymphocytes, which in turn, were more sensitive to IL-15 induced proliferation than PBT cells from normal controls. Following IL-15 mediated activation, PBT cells were capable of inducing TNF production from a macrophage cell line, from syngeneic PB monocytes, and from synovial macrophage/synoviocyte co-cultures, through a cell-contact dependent mechanism, which required no T cell cytokine synthesis. RASFT cells exhibited similar properties, which were IL-15 dependent . IL-15 upregulated CD69 expression on CD45ROT cells and neutralisation studies determined that such CD69 expression, in combination with LFA-1 and ICAM-1, was partly responsible for cell-contact mediated macrophage activation by T cells. Finally, in a murine model, IL-15 injection induced significant local tissue T cell invasion, confirming previous observations of its chemotactic properties. IL-15 expression in RA synovial membrane therefore provides a mechanism whereby polyclonal T cell recruitment and activation can lead to macrophage activation and TNF production, without T cell cytokine synthesis