Ehlers-Danlos syndrome VIII with novel C1R variant accompanying white matter changes

Ehlers-Danlos syndrome (EDS) VIII is an autosomal dominant inherited connective tissue disorder characterized by intractable periodontal inflammation, absence of gingiva, pretibial plaques, skin hyperextensibility, joint hypermobility, and tissue fragility with onset in the childhood or adolescence. In a recent report, heterozygous variants of the C1R or C1S related to the classical complement pathway were identified in families with history of EDS VIII. The current report describes a Korean 34-year-old female carrying a novel missense variant of C1R c.925T>G (p.Cys309Gly) and exhibiting early severe periodontitis, skin fragility, and joint hypermobility. The patient also had frontal, parietal, and temporal white matter brain lesions without definite vascular abnormalities on brain magnetic resonance imaging, which have not been surveyed meticulously in EDS VIII. Considering the genetic alteration of classic complement pathways in this condition, it is necessary to carefully observe multisystemic inflammation processes such as changes in brain white matter.

DNA Microarray-Based Gene Expression Profiling in Porcine Keratocytes and Corneal Endothelial Cells and Comparative Analysis Associated with Xeno-related Rejection

Porcine to rat corneal xenotransplantation resulted in severe inflammation and rejection of the corneal stroma, whereas an allograft showed mainly endothelial cell-associated rejection. We, therefore, investigated and compared the gene expression between porcine keratocytes and corneal endothelial cells. RNA was isolated from primary cultured porcine or human keratocytes and porcine corneal endothelial cells. Gene expression was comparatively analyzed after normalization with microarray method using Platinum pig 13 K oligo chip (GenoCheck Co., Ltd., Ansan, Korea). Real-time polymerase chain reaction (PCR) was performed for C1R, CCL2, CXCL6, and HLA-A in porcine keratocytes and corneal endothelial cells. As a result, upregulated expression more than 2 folds was observed in 1,162 genes of porcine keratocytes versus porcine endothelial cells. Among the immune-regulatory genes, SEMA3C, CCL2, CXCL6, F3, HLA-A, CD97, IFI30, C1R, and G1P3 were highly expressed in porcine keratocytes, compared to porcine corneal endothelial cells or human keratocytes. When measured by real-time PCR, the expression of C1R, CCL2, and HLA-A was higher in porcine keratocytes compared to that in porcine corneal endothelial cells. In conclusion, the increased expression of C1R, CCL2, and HLA-A genes in porcine keratocytes might be responsible for the stromal rejection observed in a porcine to rat corneal xenotransplantation.

Obtención de un preparado del componente C1 (q, r, s) del sistema complemento / Obtaining a preparation of C1 component (q, r, s) of the complement system

Se obtuvo un preparado concentrado del componente C1 (q, r, s) del sistema completo en forma inactiva por un método de precipitación, libre de contaminación con los restantes componentes iniciales de la vía clásica, el C2 y el C4. Este preparado se mantuvo estable a 2 °C y en los ensayos funcionales fue capaz de fijarse al complejo EAC4, activar los componentes C2 y C4 para formar la convertasa del C3, y provocar la lisis de estas células. La reacción de hemólisis fue activada por el C2 e inhibida por el Cl inhibidor, lo que indica la especificidad del C1 obtenido.Este pudiera emplearse para lograr un antisuero específico, para la obtención de los subcomponentes C1q, C1r y C1s, o para la estandarización de métodos funcionales de estudiar el C1, el C2 o el C1 inhibidor.

Purificación de C1r a partir de la fracción I de Cohn / Purification of C1r starting from Chron's fraction I

Se describe un método de purificación del componente C1r del sistema complemento a partir de la fracción I de Cohn mediante cromatografías secuenciales en Bio Rex 70, DEAE Sephacel y Sephacryl S 200. La presencia de C1r en las fracciones eluidas en las diferentes corridas cromatográficas se detectó por nefelometría. La pureza del C1r obtenido se determinó por inmunoelectroforesis contra un suero anti proteínas séricas humanas y un antisuero específico. A partir del C1r purificado se produjo en conejos un antisuero de buena calidad, útil para la cuantificación del C1r sérico