Psoriasis is a type I-deviated disease characterized by the presence of interferon (IFN)-␥ and multiple IFN-related inflammatory genes in lesions. Because interleukin (IL)-23 is now recognized to play a role in the recruitment of inflammatory cells in a T helper cell (Th)1-mediated disease, we examined psoriasis skin lesions for production of this newly described cytokine. IL-23 is composed of two subunits: a unique p19 subunit and a p40 subunit shared with IL-12. We found a reliable increase in p19 mRNA by quantitative reverse transcription polymerase chain reaction in lesional skin compared with nonlesional skin (22.3-fold increase; P ϭ 0.001). The p40 subunit, shared by IL-12 and IL-23, increased by 11.6-fold compared with nonlesional skin (P ϭ 0.003), but the IL-12 p35 subunit was not increased in lesional skin. IL-23 was expressed mainly by dermal cells and increased p40 immunoreactivity was visualized in large dermal cells in the lesions. Cell isolation experiments from psoriatic tissue showed strong expression of p19 mRNA in cells expressing monocyte (CD14 ϩ CD11c ϩ CD83 Ϫ ) and mature dendritic cell (DC) markers (CD14 Ϫ CD11c ϩ CD83 ϩ ), whereas in culture, the mRNAs for p40 and p19 were strongly up-regulated in stimulated monocytes and monocyte-derived DCs, persisting in the latter for much longer periods than IL-12. Our data suggest that IL-23 is playing a more dominant role than IL-12 in psoriasis, a Th1 type of human inflammatory disease.

ABSTRACTcDNA clones encoding human factor V have been isolated from an oligo(dT)-primed human fetal liver cDNA library prepared with vector Charon 21A. The cDNA sequence of factor V from three overlapping clones includes a 6672-base-pair (bp) coding region, a 90-bp 5' untranslated region, and a 163-bp 3' untranslated region within which is a poly(A) tail. The deduced amino acid sequence consists of 2224 amino acids inclusive of a 28-amino acid leader peptide. Direct comparison with human factor VIII reveals considerable homology between proteins in amino acid sequence and domain structure: a triplicated A domain and duplicated C domain show -40% identity with the corresponding domains in factor VIII. As in factor VIII, the A domains of factor V share -40% amino acid-sequence homology with the three highly conserved domains in ceruloplasmin. The B domain of factor V contains 35 tandem and m9 additional semiconserved repeats of nine amino acids of the form Asp-Leu-Ser-Gln-Thr-Thr/Asn-LeuSer-Pro and 2 additional semiconserved repeats of 17 amino acids. Factor V contains 37 potential N-linked glycosylation sites, 25 of which are in the B domain, and a total of 19 cysteine residues.Factor V is a large and asymmetric glycoprotein that circulates in plasma and is an essential component of the blood coagulation cascade (1, 2). During coagulation, the procofactor factor V is converted to the active cofactor, factor Va, via limited proteolysis by a-thrombin (3-5). Factor Va is a cofactor for the seine protease factor Xa, and together, factors Va and Xa assemble on a cellular or phospholipid surface with divalent metal ions to form the prothrombinase complex (1, 6-11). This complex enhances factor Xa activity -350,000 fold. The prothrombinase complex is analogous to another complex that proteolytically cleaves zymogen factor X to active enzyme factor Xa-this other "ten-ase" complex is composed of a serine protease (factor IXa), a cofactor (factor VIIMa), phospholipid, and calcium (13, 14).In addition to the similarities between serine proteases (factors Xa and IXa) and in overall enzyme complex architecture, the cofactors (factor Va and factor VIIIa) are very similar proteins structurally and functionally (13-16). Heavy and light chains of bovine factor Va and porcine factor VIIIa possess amino acid-sequence homology at the amino-terminal portion of the chains-regions of homology that are also homologous to regions in the triplicated domain structure of ceruloplasmin, the primary transport protein for copper in plasma. Available data therefore suggest that factor V, factor VIII, and ceruloplasmin are members of a family of structurally related proteins (15).The molecular cloning and sequencing of human factor VIII and human ceruloplasmin gives evidence for a common domain structure and has enabled detailed comparison of their structures (16,(17)(18)(19)(20). Recently Kane and Davie (21) published a partial cDNA sequence for human factor V that coded for "40% of the molecule. This cDNA coded for the light chain and a small ...

Factor VIII (antihemophilic factor) is a high molecular weight plasma glycoprotein that participates in the blood clotting cascade. The recent cloning and sequence analysis of the cDNA encoding human factor VIII revealed an obvious domain structure for the protein, which can be represented as Al-A2-B-A3-Cl-C2. We now report the DNA sequence analysis of porcine exons encoding the entire B domain and part of the A2 and A3 domains. We found an unusually high degree of porcine-human amino acid sequence divergence in the B region compared with the limited sequence available for other regions of the porcine factor VIII molecule. In addition to sequence divergence, there are numerous gaps in the porcine B domain totalling over 200 amino acids. Recombinant DNA techniques were used to effect the removal of large segments of DNA encoding the B domain from the full-length human factor VIII cDNA. These constructs directed the synthesis of biologically active factor VIII when introduced into mammalian cells despite the deletion of up to 38% ofthe factor VIII molecule.Hemophilia A is a bleeding disorder resulting from a deficiency or abnormality of the blood clotting protein, factor VIII (for review, see ref. 1). Factor VIII functions in the blood clotting cascade as the cofactor for factor IXa proteolytic activation offactor X. The blood clotting pathway in which factor VIII participates eventually results in the proteolytic cleavage of fibrinogen to form insoluble fibrin polymers. In vivo, fibrin deposition in conjunction with platelet aggregation act to curtail blood loss from a damaged vessel.The cofactor activity of factor VIII acts to increase the Vm,, of the factor IXa-dependent activation of factor X by at least 4 orders of magnitude. Factor VIII does not function proteolytically in this reaction but can itself be proteolytically activated by other coagulation enzymes such as factor X or thrombin. Neither the mechanism of factor VIII activation nor the nature of its cofactor activity is well understood.Recently the entire human factor VIII gene, spanning over 185 kilobases (kb) of the X chromosome, and full-length cDNA have been cloned (2-5). The DNA sequence of the cDNA and NH2-terminal amino acid sequence analysis of the plasma protein shows factor VIII to be synthesized as a single-chain precursor of 2351 amino acids from which a 19-amino acid "signal sequence" is cleaved during translation. A computer-aided search for factor VIII intramolecular homologies revealed three distinct structural domains, including a triplicated region (A domain) of -330 amino acids, a duplicated region (C domain) of '=150 amino acids, and a unique region (B domain) of -980 amino acids. These domains are arranged in the order A1-A2-B-A3-C1-C2 (see In this report, we present DNA sequence analysis that demonstrates a very high degree of divergence between porcine and human B domains of the factor VIII gene. In addition, removal of the majority of DNA encoding the B domain from a full-length human factor VIII cDNA is not detrimental...

Introduction The receptor for advanced glycation end products (RAGE), a multi-ligand member of the immunoglobulin superfamily, contributes to acute and chronic disease processes, including sepsis.

IL-22 is made by a unique set of innate and adaptive immune cells, including the recently identified noncytolytic NK, lymphoid tissue-inducer, Th17, and Th22 cells. The direct effects of IL-22 are restricted to nonhematopoietic cells, its receptor expressed on the surface of only epithelial cells and some fibroblasts in various organs, including parenchymal tissue of the gut, lung, skin, and liver. Despite this cellular restriction on IL-22 activity, we demonstrate that IL-22 induces effects on systemic biochemical, cellular, and physiological parameters. By utilizing adenoviral-mediated delivery of IL-22 and systemic administration of IL-22 protein, we observed that IL-22 modulates factors involved in coagulation, including fibrinogen levels and platelet numbers, and cellular constituents of blood, such as neutrophil and RBC counts. Furthermore, we observed that IL-22 induces thymic atrophy, body weight loss, and renal proximal tubule metabolic activity. These cellular and physiological parameters are indicative of a systemic inflammatory state. We observed that IL-22 induces biochemical changes in the liver including induction of fibrinogen, CXCL1, and serum amyloid A that likely contribute to the reported cellular and physiological effects of IL-22. Based on these findings, we propose that downstream of its expression and impact in local tissue inflammation, circulating IL-22 can further induce changes in systemic physiology that is indicative of an acute-phase response.