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Trachsel, Eveline; Bootz, Frank; Silacci, Michela; Kaspar, Manuela; Kosmehl, Hartwig; Neri, Dario

doi:10.1186/ar2115

Cited by 70 publications

(33 citation statements)

References 51 publications

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“…We are currently investigating whether the presence of UG is able to enhance the anti-inflammatory properties of similar cytokines. In fact, it has recently been shown that an immunocytokine made up of the scFv L19 and IL10 selectively accumulates at sites of arthritis in collagen-induced arthritis experimental animal models and that it inhibits disease progression (38). The activity of this anti-inflammatory immunocytokine could be further enhanced using UG, which, again, per se possesses antiinflammatory properties and, in addition, yields a stable covalently linked homodimer.…”

Section: Discussionmentioning

confidence: 99%

Use of Uteroglobin for the Engineering of Polyvalent, Polyspecific Fusion Proteins

Ventura

Sassi

Fossati

et al. 2009

Journal of Biological Chemistry

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We report a novel strategy to engineer and express stable and soluble human recombinant polyvalent/polyspecific fusion proteins. The procedure is based on the use of a central skeleton of uteroglobin, a small and very soluble covalently linked homodimeric protein that is very resistant to proteolytic enzymes and to pH variations. Using a human recombinant antibody (scFv) specific for the angiogenesis marker domain B of fibronectin, interleukin 2, and an scFv able to neutralize tumor necrosis factor-␣, we expressed various biologically active uteroglobin fusion proteins. The results demonstrate the possibility to generate monospecific divalent and tetravalent antibodies, immunocytokines, and dual specificity tetravalent antibodies. Furthermore, compared with similar fusion proteins in which uteroglobin was not used, the use of uteroglobin improved properties of solubility and stability. Indeed, in the reported cases it was possible to vacuum dry and reconstitute the proteins without any aggregation or loss in protein and biological activity.The generation of recombinant polyvalent and/or polyspecific fusion proteins for use as components of novel drugs is still hindered by factors that limit their production, storage, and use, chief of which are issues related to instability and/or inadequate solubility. Here we describe a novel approach based on the use of uteroglobin (UG) 3 as a skeleton for the generation of polyvalent/polyspecific recombinant proteins. Human UG is a small (15.8 kDa) globular, nonglycosylated, and homodimeric secreted protein that was discovered independently by two groups in the 1960s in rabbit uterus (1, 2), and it is the first member of a new superfamily of proteins, the so-called Secretoglobins (Scgb) (3). UG is present in the blood at a concentration of about 15 g/ml and is found in urine and in other body fluids. The UG monomer is composed of about 70 amino acids, depending on the species, and is organized in a four ␣-helix secondary structure; the two subunits are joined in an antiparallel fashion by disulfide bridges established between two highly conserved cysteine residues in amino-and carboxyl-terminal positions (4) (see Fig. 1). The exact functions of UG are not yet clear, but the protein has been reported to have antiinflammatory properties due to its ability to inhibit the soluble phospholipase A 2 . Moreover, UG contains a central hydrophobic cavity able to accommodate hydrophobic molecules such as progesterone, retinol, and prostaglandin D 2 . Theoretically, this cavity could be loaded with different types of therapeutic hydrophobic substances and delivered to targets (for exhaustive reviews on UG, see Refs. 5, 6and references therein).The high solubility and stability of UG to pH and temperature variations, its resistance to proteases, and its homodimeric structure prompted us to consider the protein as a candidate linker for the generation of polyvalent and polyspecific recombinant proteins. We demonstrate here that the use of UG as a linker could provide a general method for...

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Section: Discussionmentioning

confidence: 99%

Use of Uteroglobin for the Engineering of Polyvalent, Polyspecific Fusion Proteins

Ventura

Sassi

Fossati

et al. 2009

Journal of Biological Chemistry

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“…Constructs containing promotors regulated by doxycycline or tetracycline were already effective in experimental arthritis [64]. Apart from gene therapy, other strategies for IL-10 targeting were studied as described by Trachsel et al, [68] where an antibody was fused with IL-10 (L19-IL-10) which accumulates at the inflamed joint and displayed a therapeutic activity [68].…”

Section: Il-10 and Rheumatoid Arthritismentioning

confidence: 99%

“…These scaffolds which release IL-10 very slowly, could be seeded with autologous chondrocytes and then implanted. The use of osmotic minipumps which secrete continuously IL-10 or an antibody mediated release of it [68,131] may be combined with gene therapeutic approaches.…”

Section: Conclusion and Future Prospectsmentioning

confidence: 99%

Interleukin-10 and Articular Cartilage: Experimental Therapeutical Approaches in Cartilage Disorders

Schulze‐Tanzil¹,

Zreiqat²,

Sabat³

et al. 2009

CGT

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Interleukin (IL)-10 is a well known anti-inflammatory and immunoregulatory cytokine, mainly released by, and acting on cells of the immune system such as monocytes, macrophages, T cells, NK cells, and B cells. IL-10 is also produced by a few connective tissue cell types including chondrocytes and is involved in processes such as connective tissue extracellular matrix remodelling, although it's exact function in articular cartilage remains unclear. This review article summarizes after a short insight into functions of IL-10 in the immune system most of the published literature on the role of IL-10 in articular cartilage homeostasis and disorders. A critical analysis of the present literature was undertaken leading to a survey of the significance of IL-10 in rheumatoid arthritis (RA), osteoarthritis (OA) and blood induced cartilage damage with particular focus on the direct impact of IL-10 on chondrocyte biology. IL-10 is up-regulated in RA and OA and therapeutic effects of IL-10 in experimental arthritis using several gene therapeutic approaches were reported, mainly through an immune cell mediated immunosuppression mechanism. Recently, a direct anti-inflammatory, -catabolic and -apoptotic potential of IL-10 in cartilage was described, suggesting a chondroprotective effect of IL-10, not only in RA and OA, but also in non-RA and non-systemic cartilage disorders. Chondroprotection by IL-10 may be a promising tool in arthritis therapy, as IL-10 plays a role in joint and cartilage immunoregulation and homeostasis. However, a crucial problem remains to be the optimisation of local and continuous therapeutic effective levels of IL-10 administration in the joint.

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“…In recent clinical trials, the bacterial expression of the IL-10 transgene has shown to be biologically contained and, the treatment safe and effective in Chron's disease patients [46]. Other proposed approaches to target specific tissues are the use of fusion proteins of IL-10 and monoclonal antibodies specific for the targeted cell type, that has been shown to be effective in an experimental model of rheumatoid arthritis [48], and the chemical alteration of IL-10. IL-10 modified with mannose-6 phosphate shows a prevalent homing to the liver, in particular to fibrogenic hepatic stellate cells expressing the mannose 6 phosphate/insulin-like growth factor II receptor, and could be applied to the treatment of cirrhosis [49].…”

Section: Il-10 and Il-10 Agonistsmentioning

confidence: 99%

Caveats Regarding the Use of IL-10 and IL-10 Antagonist as Immunotherapeutic Factors

Colino¹

2007

LDDD

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Untitled

Cited by 70 publications

References 51 publications

Use of Uteroglobin for the Engineering of Polyvalent, Polyspecific Fusion Proteins

Use of Uteroglobin for the Engineering of Polyvalent, Polyspecific Fusion Proteins

Interleukin-10 and Articular Cartilage: Experimental Therapeutical Approaches in Cartilage Disorders

Caveats Regarding the Use of IL-10 and IL-10 Antagonist as Immunotherapeutic Factors

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