Establishment and validation of OX40/FcγR-humanized mice for the study of agonistic anti-OX40 antibody / 上海交通大学学报（医学版）

Objective • To establish a rapid method to evaluate the activity of agonistic antibody using OX40 (tumor necrosis factor receptor superfamily member 4)/FcγR (Fcγ receptor)-humanized mice. Methods • Bone marrow cells from OX40-humanized mice and FcγR-humanized mice were collected and mixed with equal ratio. Then the mixed bone marrow cells were administrated into irradiated wild-type mice through the tail veins. The reconstruction efficiency of the immune system was confirmed by detecting the expression of hOX40 and hFcγR in the immune cells of chimera mice. After the chimera mice were generated successfully, they were used to evaluate the immunostimulatory activity of anti-hOX40 antibodies to CD4+ or CD8+ T cells. The results of flow cytometry were statistically analyzed. The unpaired t-test was used to compare the means between the two groups, and oneway ANOVA was used to compare the means between multiple groups. Results • Flow cytometry analysis showed that wild-type recipient mice were efficiently reconstituted with hFcγR expressing cells and hOX40 expressing cells to generate OX40/FcγR-humanized bone marrow chimera mice. In these mice, B cells and myeloid cells expressed hFcγRs (P<0.05), and T cells expressed hOX40 upon in vitro stimulation (P<0.05). When these mice were used to evaluate the immunostimulatory activity of anti-hOX40 antibody, significant expressions of IFN-γ and hOX40 were observed (P<0.05). Conclusion • OX40/FcγR-humanized bone marrow chimera mice are generated based on hFcγR expressing cells and hOX40 expressing cells, suggesting a rapid method to build a mouse model with both hFcγR and hOX40 expression. These mice are suitable for evaluating the immunostimulatory activity of agonistic human anti-hOX40 antibodies.

Study of tumor necrosis factor receptor superfamily 1B gene polymorphism in relation to the outcomes of HCV infection / 中华肝脏病杂志

Objective@#To investigate the tumor necrosis factor receptor superfamily 1B gene (TNFRSF1B) polymorphism in relation to the outcomes of hepatitis C virus (HCV) infection.@*Methods@#One thousand six hundred and forty-five cases without HCV infection, 545 cases with HCV clearance, and 783 cases with chronic HCV infection were enrolled. TaqMan probe method was used to investigate genotype rs1061622 (T > G) and rs1061624 (G > A). Two single nucleotide polymorphisms (SNPs) sites were genotyped and haplotypes were constructed to evaluate their relation with the outcome of HCV infection.@*Results@#Logistic regression analysis showed that there was no relation to the two SNPs with HCV infection susceptibility and chronicity (P > 0.05). Haplotype analysis showed that carrier TA had an increased susceptibility to HCV infection [adjusted odds ratio (OR) = 1.15, 95% confidence interval (CI): 1.01 to 1.30, P = 0.038)]. Carrier TA and GG haplotypes were conducive to chronic HCV infection (adjusted OR = 1.28, 95% CI: 1.08 to 1.53, P = 0.006; OR = 1.31, 95% CI: 1.03 to 1.66, P = 0.026).@*Conclusion@#The combinational effects of rs1061622 and rs1061624 in TNFRSF1B gene may increase the risk of HCV chronicity and infection.

Association between TNFRSF11A and TNFRSF11B gene polymorphisms and the outcome of hepatitis C virus infection / 中华流行病学杂志

Objective@#To explore the relationship between the tumor necrosis factor receptor superfamily members 11A (TNFRSF11A) and 11B (TNFRSF11B) gene polymorphisms and the outcome of hepatitis C virus (HCV) infection.@*Methods@#In this case-control study, 749 cases of persistent HCV infection, 494 cases of spontaneous clearance and 1 486 control subjects were included from 2008 to 2016. TaqMan-MGB probe method was used to detect the genotype of TNFRSF11A rs1805034 and TNFRSF11B rs2073617. The genotypes distribution of the two single nucleotide polymorphisms (SNP) were analyzed in different populations.@*Results@#Co-dominant model showed that individuals carrying the rs2073617 CC genotype were prone to have chronic HCV infection, compared with individuals carrying the rs2073617 TT genotype (OR=1.517, 95%CI: 1.055-2.181, P=0.024). Recessive model results showed that individuals carrying rs2073617 CC genotype were more likely to develop chronic HCV infection compared with individuals carrying rs2073617 TT or TC genotype (OR=1.435, 95%CI: 1.033-1.996, P=0.032). Additive model showed that the risk for chronic HCV infection increased with the increase of the number of rs2073617 C alleles (OR=1.204, 95%CI: 1.013-1.431, P=0.035).@*Conclusion@#The genetic polymorphism of TNFRSF11B rs2073617 might be related with the chronicity of HCV infection.

Anti-inflammatory mechanism of Scutellarlae Radix based on network pharmacology / 中草药

Objective: To explore the anti-inflammatory mechanism of Scutellarlae Radix (SR) by the network pharmacology. Methods: Firstly, the components in SR were searched through TCMSP database and screened with “Lipinski rule” and “Oral Bioavailability > 30%” rules. The targets of above components selected by PharmMapper web server and Cytoscape 3.4.0 was used to build a network between components and targets (component-target network, CTN). Secondly, “anti-inflammatory” targets was searched from Therapeutic Target Database (TTD) with keyword “anti-inflammatory”, and targets retrieved were used to build a protein-protein interaction (PPI) network based on the analysis by String database. To obtain anti-inflammatory targets of the active components in SR, the PPI network was fused with the CTN. Finally, the DAVID database was used to perform KEGG pathway enrichment analysis in order to explore the anti-inflammatory mechanism of SR. Results: Twenty-eight components in SR were obtained, including flavonoids such as baicalin, baicalein, wogonin, wogonoside, etc, alkaloids such as berberine, and epiberberine, and phenols such as dihydromyricetin, etc. Mitogen-activated protein kinase (MAPK14), tumor necrosis factor receptor superfamily 1A (TNFRSF1A), epidermal growth factor receptor (EGFR), and E-selectin (SELE) were the main targets of SR' anti-inflammatory effect. Salvigenin, epicatechin, and astragalusine mainly acted on MAPK14; Carthamidin acted on TNFRSF1A; Dihydromubutin A, 5,7,4’-trihydroxy-8-methoxyflavone, 5,7,4’-trihydroxy-8-methoxyflavanone, baicalin, and other components mainly acted on EGFR. There were 11 KEGG pathways, mainly related to TNF signaling pathways, MAPK signaling pathway, etc. Conclusion: There are three main anti-inflammatory mechanisms in SR, which can inhibit the production of inflammatory factors, inhibit the binding of inflammatory factors to their respective receptors, and block the initiation of inflammatory reactions.

Effect of Wenhua Juanbi Recipe () on expression of receptor activator of nuclear factor kappa B ligand, osteoprotegerin, and tumor necrosis factor receptor superfamily member 14 in rats with collagen-induced arthritis / 中国结合医学杂志

<p><b>OBJECTIVES</b>To study the effect of Wenhua Juanbi Recipe (, WJR) on expression of receptor activator of nuclear factor kappa B ligand (RANKL), osteoprotegerin (OPG), and tumor necrosis factor receptor superfamily member 14 (TNFRSF14, also known as LIGHT) in rats with collagen-induced arthritis (CIA).</p><p><b>METHODS</b>CIA rats were generated by subcutaneous injection of bovine collagen type-II at the tail base. Sixty CIA rats were randomly assigned (10 animals/group) to: model, methotrexate (MTX)-treated (0.78 mg/kg body weight), and WJR-treated (22.9 g/kg) groups. Healthy normal rats (n=10) were used as the normal control. Treatments or saline were administered once daily by oral gavage. Rats were sacrifificed at day 28 post-treatment and knee synovium and peripheral blood serum were collected. Toe swelling degree and expression of RANKL, OPG, and LIGHT were determined by Western blot and immunohistochemistry.</p><p><b>RESULTS</b>Compared with the normal group, toe swelling degree was signifificantly increased in the model group (P<0.01). After treatment, toe swelling degree decreased signifificantly in the WJR and MTX groups compared with the model group (P<0.01). Compared with the normal group, expression of RANKL and LIGHT were signifificantly increased and OPG signifificantly decreased in peripheral blood and synovium of the model group (P<0.01). Conversely, RANKL and LIGHT expression were signifificantly reduced and OPG increased in the WJR and MTX groups compared with the model group (P<0.01). No statistically significant difference existed between WJR and MTX groups.</p><p><b>CONCLUSION</b>WJR likely acts by reducing RANKL expression and increasing OPG expression, thus inhibiting RANKL/RANK interaction and reducing LIGHT expression, thereby inhibiting osteoclast formation/activation to block bone erosion.</p>

Novel regulatory SNPs in the promoter region of the TNFRSF18 gene in a Gabonese population

Parasites are accountable for driving diversity within immune gene families. We identified and investigated regulatory single nucleotide polymorphisms (SNPs) in the promoter regions of the tumor necrosis factor receptor superfamily member 18 (TNFRSF18) gene by direct sequencing in a group of male Gabonese individuals exposed to a wide array of parasitic diseases such as malaria, filariasis and schistosomiasis. Two new promoter variants were identified in 40 individuals. Both novel variants were heterozygous and were linked to SNP #rs3753344 (C/T), which has been described. One of the SNP variants (ss2080581728) was close to the general transcription factor site, the TATA box. We further validated these new promoter variants for their allelic gene expression using transient transfection assays. One new promoter variant with two base changes (C/T - ss2080581728/rs3753344) displayed an altered expression of the marker gene. Both novel variants remained less active at the non-induced state in comparison to the major allele. The allele frequencies observed in this study were consistent with data for other African populations. The detection and analysis of these human immune gene polymorphisms contribute to a better understanding of the interaction between host-parasite and expression of Treg activity.