Pinçon-Raymond M, Cardot P, Thenet S. Epidermal growth factor receptor is involved in enterocyte anoikis through the dismantling of E-cadherin-mediated junctions.
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A plasmid vector carrying the immunoprotective amino-terminal fragment of Taenia solium paramyosin (VW2-1) was designed for genetic vaccination studies. Mice that were genetically immunized with VW2-1 and challenged by intraperitoneal inoculation of Taenia crassiceps cysticerci showed 43 to 48% reductions in the parasite burden, values which were similar to values obtained previously when the recombinant protein was used.Human and porcine cysticercosis caused by Taenia solium is still prevalent in several countries of Latin America, Africa, and Asia (11). A strong effort is currently being directed toward the development of an effective vaccine against porcine cysticercosis. A number of strategies have been proposed, including the use of parasite crude extracts (9, 17), recombinant proteins (2, 3, 15), synthetic peptides (4), phage display (8), and genetic immunization (10, 18). Genetic vaccines are particularly appealing for applications in developing countries as they can be inexpensive to produce and store (5). In the present study, we evaluated the use of genetic immunization with the amino-terminal fragment of T. solium paramyosin (VW2-1), which was previously shown to be protective (15), as an alternative strategy for vaccination in the murine model of Taenia crassiceps cysticercosis.A plasmid construct encoding a synthetic sequence for VW2-1 was designed for genetic immunization. The codon usage of the wild-type coding sequence for VW2-1 was adapted for mammalian cells (synVW2-1) by using procedures for gene synthesis (1). Briefly, a set of overlapping 60-mer oligonucleotides were assembled by PCR to construct synVW2-1, which was inserted into the pCMV plasmid vector (14). The construct was confirmed by DNA sequencing before large-scale purification with a QIAGEN Endofree Plasmid Giga kit (QIA-GEN). A comparison of the wild-type and synthetic sequences of VW2-1 showed substantial changes in first (11.2%), second (4.85%), and third (52.6%) positions of codons. An extensive comparison of the outcomes of the immune responses elicited in mice genetically immunized with wild-type VW2-1 and with synVW2-1 consistently showed higher antibody and cellular immune responses with the synthetic gene (data not shown). The synthetic coding sequence is available from us upon request.Vaccination assays were carried out by using 8-to 10-weekold female BALB/c mice that were bilaterally inoculated in the tibialis anterior muscle with pCMV-synVW2-1 or pCMV blank vector (100 g of DNA per mouse in 100 l of 0.9% saline). A third group contained naïve mice. After 2 and 4 weeks, mice were boosted with identical doses of plasmid DNA.In order to evaluate if the antibodies raised by genetic immunization were cross-reactive with paramyosins of T. solium and T. crassiceps, Western blot assays were carried out as described previously (16). Briefly, nitrocellulose membranes were blotted with recombinant full-length TPmy (recTPmy) and recombinant VW2-1 (recVW2-1) or with crude protein extracts from T. solium and T. crassiceps cysts (6). M...
We assessed the role of plasminogen activator inhibitor-1 (PAI-1) and matrix metalloproteinase 9 (MMP9) in wound healing process and in the bone marrow mononuclear cells (BMMNC)-related effects on physiological and pathological wound healing. A full thickness excision wound was created by removal of the skin on the midback of irradiated and nonirradiated animals. Angiogenesis and re-epithelialization were markedly increased in PAI-12/2 mice compared to wild-type (WT) animals. We revealed high MMP activity in tissue of PAI-12/2 animals. Of interest, the wound healing process was reduced in PAI-12/2:MMP92/2 animals compared to PAI-12/2 mice, suggesting a key role of MMP9 in beneficial effect of PAI-1 deficiency on wound closure. To unravel the role of PAI-1 in BMMNC relative effects, mice were treated with or without local injection of BMMNC isolated from WT, PAI-12/2, and PAI-12/2: MMP92/2 animals for 14 days (10 6 cells, n 5 6 per group). In WT nonirradiated mice, transplantation of BMMNC isolated from PAI-12/2 animals enhanced wound formation when compared with WT BMMNC. BMMNC differentiation into cells with endothelial phenotype was enhanced by PAI-1 deficiency. These effects were abrogated in PAI-12/2:MMP92/2 and MMP92/2 BMMNC. In addition, using chimeric mice, we demonstrated that PAI-1 deficiency environment increased the BMMNC-GFP recruitment to the wound site, whereas this effect was abrogated when using PAI-12/2:MMP92/2 BMMNC. PAI-1 deficiency, at least through MMP9 upregulation, enhanced wound healing and BMMNC therapeutic potential in irradiated and nonirradiated animals.
Skin wound repair requires the development of different kinds of biomaterials that must be capable of restoring the damaged tissue. Type I collagen and chitosan have been widely used to develop scaffolds for skin engineering because of their cell-related signaling properties such as proliferation, migration, and survival. Collagen is the major component of the skin extracellular matrix (ECM), while chitosan mimics the structure of the native polysaccharides and glycosaminoglycans in the ECM. Chitosan and its derivatives are also widely used as drug delivery vehicles since they are biodegradable and noncytotoxic. Regulation of the inflammatory response is crucial for wound healing and tissue regeneration processes; and, consequently, the development of biomaterials such as hydrogels with anti-inflammatory properties is very important and permissive for the growth of cells. In the last years, it has been shown that mesenchymal stem cells have clinical importance in the treatment of different pathologies, for example, skin injuries. In this paper, we describe the anti-inflammatory activity of collagen type 1/chitosan/dexamethasone hydrogel, which is permissive for the culture of human adipose-derived mesenchymal stem cells (hADMSC). Our results show that hADMSC cultured in the hydrogel are viable, proliferate, and secrete the anti-inflammatory cytokine interleukin-10 (IL-10) but not the inflammatory cytokine Tumor Necrosis Factor-alpha (TNF-α).
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