FasR x TGF-β2

Background/Aims: This study aimed to investigate the effects of the VR-10 TSP-1 synthetic polypeptide on cytokines and the proliferation and migration of endothelial cells, as well as exploring a new method for anti-ocular neoangiogenesis. Methods: We measured the proliferation of RF/6A cells by an MTT assay and investigated the migration of RF/6A cells by a Transwell chamber assay. We examined the mRNA transcript levels of TGF-β2, VEGF, PEDF, Bcl-2 and FasL in RF/6A cells by RT-PCR and evaluated the expression of Fas and caspase-3 proteins in RF/6A cells by western blot analysis. Results: 1. TSP-1 (1 µg/ml) and synthetic peptide VR-10 (0.1 µg/ml, 1 µg/ml and 10 µg/ml) inhibited the proliferation of RF/6A cells in a time and dose-dependent way. 2. TSP-1 and synthetic peptide VR-10 could inhibit the migration of RF/6A cells in a Transwell chamber (P < 0.001). It was demonstrated that 10 µg/ml synthetic peptide VR-10 had the strongest effect. 3. The expression of TGF-β2 mRNA in RF/6A cells increased after treatment with 1 µg/ml TSP-1 (P < 0.0001). However, there was no significant difference between the synthetic peptide VR-10 and the control group (P > 0.05). Expression of PEDF mRNA in RF/6A cells was increased after treatment with 1 µg/ml TSP-1 and synthetic peptide VR-10. We demonstrated that 10 µg/ml synthetic peptide VR-10 had the strongest effect (P < 0.001). There were significant differences between groups (P < 0.001). Expression of TGF-β2 mRNA in RF/6A cells increased after treatment with 1 µg/ml TSP-1 (P = 0.000). There was no significant difference between the synthetic peptide VR-10 and the control group (P > 0.05). PEDF mRNA expression in RF/6A cells decreased after 1 µg/ml TSP-1 and synthetic peptide VR-10 therapy, among which 10 µg/ml synthetic peptide VR-10 demonstrated the strongest effect (P < 0.001). There were significant differences between groups (P < 0.001), except for the 1 µg/ml synthetic peptide VR-10 and 1 µg/ml synthetic peptide VR-10 groups (P = 0.615). 4. Compared with the control group, FasL mRNA expression was significantly increased in the 10 µg/ml synthetic peptide VR-10 treatment group; however, Bcl-2 mRNA expression was decreased. 5. Western blotting showed that RF/6A cells in the control group mainly expressed the 32 kD procaspase-3 forms. For the 10 µg/ml synthetic peptide, VR-10 treatment group, it showed decreased expression of procaspase-3 (32 kD) and concomitant increased expression of its shorter pro apoptotic forms (20 kD). Compared with the control group, Fas protein expression significantly increased in the 10 µg/ml synthetic peptide VR-10 treatment group. Conclusions: Synthetic peptide VR-10 had an inhibitory action on the proliferation and migration of RF/6A cells. VR-10 inhibited angiogenesis by its combined actions, which included up-regulating the expression of an anti-angiogenesis gene, namely, pigment epithelium-derived factor (PEDF), down-regulating the expression of the pro-angiogenic vascular endothelial growth factor (VEGF), and mediated endothelial cell apoptosis.

We aimed to develop multiple single-nucleotide polymorphism (SNP)-based risk models associated with the risk of transplant outcomes including graft-versus-host disease (GVHD).

The study evaluated 259 SNPs in 53 genes in 394 pairs of donors and recipients. In a discovery set (n=307 receiving related donor transplantation), overall survival, relapse-free survival (RFS), nonrelapse mortality, and acute or chronic GVHD were evaluated.

Eight recipients' SNPs of IL2, IL6R, FAS, EDN1, TGFB1, and NFKBIA genes and 12 donors' SNPs of NOS1, IL1B, TGFB2, NOD2/CARD15, TNFRII, IL1R1, and FCGR2A genes were identified in univariate analyses. Risk models were generated using significant clinical variables and genetic SNP markers after filtering out through multivariate analyses. Then, we divided patients into four quartiles (25%, Q) according to their risks. The final models stratified patients into low-risk (Q1), moderate-risk (Q2, Q3), and high-risk (Q4) groups in terms of overall survival (P<0.0001), RFS (P<0.0001), nonrelapse mortality (P=0.0043), and acute GVHD (P<0.0001), but not for chronic GVHD (P=0.763). External validation was performed in 87 transplant pairs that received matched unrelated donor transplantation, especially for RFS (P=0.016) and acute GVHD (P=0.027).

Risk models can improve prognostic stratification of patients according to their risk for transplant outcome.

Citation Yayi H, Danqing W, Shuyun L, Jicheng L. Immunologic Abnormality of Intrahepatic Cholestasis of Pregnancy. Am J Reprod Immunol 2010; 63: 267–273Problem  Intrahepatic cholestasis of pregnancy (ICP) is a pregnancy risk because of the possibility of pre‐term delivery and sudden intrauterine fetal death. Its pathogenesis is still under discussion.Method of study  The analysis of the recent findings on the complex immunologic events that occur in ICP were performed.Results  In ICP, an increase of type 1 cytokine (TNF‐α, IFN‐γ) associated with a decrease of type 2 cytokine (IL‐4). The decreased production of the suppressor cytokine TGF‐β2 may increase the type 1 cytokine. Fas appeared to be increased and FasL appeared to be decreased in syncytiotrophoblasts of ICP. The human leukocyte antigen gene (HLA‐G, E) in extravillous trophoblasts of ICP were significantly decreased.Conclusion  Th1/Th2 cytokine balance and HLA play important roles in the tolerance and maintenance of pregnancy. ICP may be resulting from breach of the maternal fetal immune tolerance during pregnancy.