Donor-Specific Anti-HLA Antibodies in Organ Transplantation: Transition from Serum DSA to Intra-Graft DSA

In the field of organ transplantation, donor-specific anti-HLA antibodies (DSA) have gained more popularity, as antibody-mediated rejection (AMR) has been recognized as an important factor to determine allograft survival. Thus, it is reasonable to believe that appropriate control of DSA is directly linked to well-managed immunosuppression, resulting in free from AMR. First, in order to prevent and manage AMR, it is of vital importance to be familiar with updated knowledge regarding crossmatch test and DSA detection methods, including intra-graft DSA. Second, it is also crucial to understand the standard criteria to diagnose AMR. Although pathological diagnosis and serum DSA (s-DSA) detection play the central role, the recent trend seems to be detection of intra-graft DSA (g-DSA). Third, regarding organ transplantation between sensitized pairs, the acceptable outcomes are obtained owing to recent preoperative desensitization protocols: depletion/modification of B cells, apheresis for antibodies, and inhibition of reaction between DSA and HLA. Finally, we would like to discuss the treatment of AMR. Further advances in diagnosis methods and emergences of effective treatments would be expected for acceptable control of AMR. In this chapter, we will review from the basics to recent topics in order to understand DSA and AMR

Japón: servicio universal, alta definición y movilidad

“Diversidad cultural y audiovisual: buenas prácticas e indicadores” (ref. CSO2011-26241), pertenece al Plan Nacional I+D+i del Ministerio de Economía y Competitividad de España. “Convergencia digital: el futuro de las tecnologías y los contenidos de la información y la comunicación”, está financiado por la Comisión de Apoyo al Personal de Educación Superior (CAPES), de Brasil, y cuenta con la participación de investigadores de las universidades Carlos III de Madrid, do Vale do Rio dos Sinos y Federal de Sergipe

Cytoskeletal inhibitors, anti-adhesion molecule antibodies, and lectins inhibit hepatocyte spheroid formation.

We investigated the role of cytoskeletons, adhesion molecules, membrane-glycosylations, and proteoglycans in forming the shape of adult rat hepatocyte spheroids. Isolated hepatocytes were cultured on dishes coated with chondroitin sulfate phosphatidyl ethanolamine (CS-PE). Spheroid-forming ability was observed after adding cytoskeletal inhibitors (cytochalasin D, colchicine, okadaic acid, mycalolide B), anti-adhesion molecule antibodies (anti-E-cadherin, anti-connexin 32, anti-zo-1), a glycosphingolipid synthetic inhibitor (N-butyldeoxynojirimycin), a proteoglycan synthetic inhibitor (p-nitrophenyl-beta-D-xylopyranoside), and several lectins. Localization of actin was studied using confocal microscopy after rhodamine-phalloidin staining. Adding cytoskeletal inhibitors on the initial day resulted in weakly clustered cell aggregates rather than smoothly formed spheroids. These effects disappeared at lower reagent concentrations. When reagents were added on day 3, after the formation of spheroids, only mycalolide B was associated with an irregular spheroid surface; the others had no effect. Adding the anti-E-cadherin, anti-connexin 32 on the initial day showed inhibition of spheroid formation, but anti-zo-1 and proteoglycan synthetic inhibitor had no effects. Among the several lectins, only Wheat Germ Agglutinin (WGA), Ricinus communis Agglutinin I (RCA-I), and Concanavalin A (ConA) showed inhibition. These results suggest that cytoskeletal conformation and some adhesion molecules are necessary to form spheroids. Based on the interactions between lectins and hepatocytes in the present study, hepatocytes appear to contain an N-linked complex or N-linked hybrid glycosylated chains

Characterization of anti-herpes simplex virus type 1 activity of an alkaloid FK 3000 from Stephania cepharantha

A morphinane alkaloid FK 3000 (6,7-di-O-acetylsinococuline) from the root tubers of Stephania cepharantha showed antiviral activity against acyclovir (ACV)- and phosphonoacetic acid (PAA)-resistant herpes simplex virus type 1 (HSV-1), influenza virus, measles virus, and poliovirus. The anti-HSV action of FK 3000 was assessed in comparison with that of PAA that inhibits the activity of HSV DNA polymerase and HSV DNA synthesis. FK 3000 inhibited the growth of thymidine kinase-deficient and ACV and PAA-resistant HSV-1 strains, as well as wild type HSV strains in Vero cells. This compound, as well as PAA, interfered with the synthesis of late viral proteins but not early viral proteins. The analysis of HSV DNA synthesis by slot blot hybridization showed that FK 3000 inhibited the viral DNA synthesis in a dose-dependent manner. However, the viral RNA was partially synthesized in the presence of FK 3000 (even at a dose that HSV DNA synthesis was inhibited) and PAA, indicating that FK 3000, as well as PAA, allowed early viral RNA synthesis but not viral DNA synthesis. Since partially purified HSV DNA polymerase activity was not inhibited by FK 3000, this compound was suggested to inhibit HSV DNA synthesis by a mechanism different from that of PAA. Stephania cepharantha(タマザキツヅラフジ)から得たモルフィン骨格を有するアルカロイドFK3000はacyclovirやphosphonoacetic acid(PAA)抵抗性を有するHSV-1, influenza virs, measles virus, poho virsに対しても抗ウイルス作用を有していた。この抗HSV作用をHSV DNA polymeraseを阻害することによりHSV DNA合成を阻害することが知られているPAAとの比較から検討した。HSVに感染したVero細胞においてFK3000は,PAAと同様に後期ウイルス蛋白の合成を阻害したが初期ウイルス蛋白には影響しなかった。Slot blot hybridization法でHSV DNA合成を調べると,FK3000は濃度依存的にウイルスDNA合成を阻害することが判明した。しかし,ウイルスRNA合成はHSV DNA合成が阻害される濃度でもFK3000およびPAAによって部分的にのみ阻害された。このことはPAAと同様にFK3000はウイルスDNA合成は阻害するが初期ウイルスRNAの合成は許容することを示している。FK3000は粗精製したHSV-DNA polymerase活性を阻害しないことから,PAAと異なった機構でHSV DNA合成を阻害していることが示唆された

Endothelium-dependent vasodilator effect of tannin extract from Cinnamonomi Cortex on isolated rat aorta

Cinnamonomi Cortex (the bark of Cinnamomum cassia BLUME) is a crude drug that is widely used in spices and medical products. Although improvement of blood flow by this plant component has long been known, there have been no reports concerning the mechanism involved. We studied the vasodilator actions of this drug especially focusing on the role of endothelium in the isolated vascular bed. Tannin from Cinnamonomi Cortex (TCC) relaxed prostaglandin F_-precontracted ring preparations of rat aorta with intact endothelium. TCC did not cause relaxation of specimens without endothelium, and TCC-induced relaxation was inhibited by pretreatment with 10^M N^G-nitro-l-arginine methyl ester. Dimer, trimer, tetramer, and pentamer components of TCC also produced endothelium-dependent vasodilatation. Stronger relaxation was caused by higher molecular weight tannins, and endothelium-dependent vasodilation even appeared at low concentrations. In conclusion, we found that TCC exhibits an endothelium-dependent vasodilatation in the isolated rat aorta mainly via endothelium derived NO. NO mediated endothelium-dependent relaxation seems to be more potent for TCC with higher molecular weight than that with lower molecular weight. 桂皮の血流改善作用については古くから知られており,これに関連した報告はあるものの,その詳細な検討はなされていない。今回我々はマグヌス法を用いて,ラット胸部大動脈輪状標本における桂皮含有クンニンの血管作動性について検討した。桂皮含有タンニンは,プロスタグランディンF_(PGF_)の血管収縮に対し,内皮保存血管において濃度依存性に血管弛緩作用が認められた。しかし,内皮除去血管及ぴN^G-nitro-l-argininemethyl ester(L-NAME)前処置内皮保存血管においては,血管弛緩作用はほぼ消失した。以上より,桂皮含有タンニンの血管弛緩作用は内皮依存性であることが明らかとなった。桂皮含有タンニンをさらに二量体から五量体までのタンニン画分に分取し検討したところ,二量体以上の重合したタンニンにおいて血管弛緩作用が認められた。また,重合度が増すに従い血管弛緩作用はより低い濃度で発揮され,作用も増強されることが明らかとなった