目的 探讨人白细胞介素(IL)-37慢病毒表达载体构建和包装.方法 以pcDNA3.1 (+)-IL-37为模板,用特异性引物聚合酶链式反应(PCR)扩增出IL-37全长编码序列;与载体pLVX-IRES-Green1酶切并连接,大量扩增纯化重组质粒及包装质粒;以适当比例共同转染293T细胞生产慢病毒;收集并浓缩慢病毒,检测慢病毒滴度及IL-37的表达.结果 所得病毒的滴度达到5×1012TU/ml;病毒感染A549细胞后,可以在细胞内稳定表达出IL-37 mRNA.结论 本实验成功构建出IL-37慢病毒表达载体,并包装获得了高滴度的病毒上清.所包装病毒上清具有感染力,在真核细胞中可以表达IL-37.

Interleukin (IL)-37 is a novel member of the IL-1 cytokine family. However, as a result of lacking efficient method to generate relatively large quantity of IL-37, little is known of its functions in man. In the present study, the recombinant human IL-37b containing a C-hexahistidine tag was expressed in Escherichia coli (E. coli). The expression level of IL-37b in E. coli was very high after induction with IPTG. Furthermore, IL-37b protein was largely found in the soluble fraction. The expressed protein was readily purified by one-step immobilized metal-ion affinity chromatography using Ni(2+)-nitrilotriacetic acid agarose. The purified IL-37b appeared as a single band on SDS-PAGE and the purity was more than 97%. The yield was 90mg IL-37b from 1l of bacterial culture. Western blotting and N-terminal sequencing confirmed the identity of the purified protein. The purified IL-37b inhibited significantly the release of tumor necrosis factor-α and IL-1β in lipopolysaccharide-activated THP-1 cells. Thus, this method provides an efficient way to obtain an active IL-37 with high yield and high purity.

BACKGROUND:The environmental pollution is one of the factors contributing to the decrease of sperm quality for human beings. The aim of this study was to assess cadmium (Cd), chromium (Cr), and copper (Cu) concentration of man in environmental pollution site, and explore relationships between men exposure to Cd, Cr, and Cu and semen-quality parameters in environmental pollution site. METHODS:Ninety five men were recruited through pollution area and controls in 2011. We measured semen quality using Computer-aided Semen Quality Analysis, and Cd, Cr, and Cu levels in seminal plasma using Graphite Gurnace Atomic Absorption Spectroscopy. Spearman rank correlation analysis was used to evaluate the correlation between Cd, Cr and Cu concentration in seminal plasma and semen quality. RESULTS:The mean of seminal plasma Cd, Cr, and Cu values in pollution area was higher than the controls. Seminal plasma Cr values displayed a significant negative correlation with total motility and normomorph sperm rate. Seminal plasma Cu values also displayed a negative correlation with normomorph sperm rate. CONCLUSIONS:Male reproductive health may be threatened by environmental pollution, and it may be influence local population diathesis.

Objective:To predict the secondary structure and B-cell epitope of human IL-37. Methods: Based on IL-37b amino acid sequence,the secondary structure was predicted by SOPMA; hydrophilicity,flexibility,accessibility index were predicted by software of ProScale,Bcepred,respectively. Combined the results according to these methods,the B cell epitopes of IL-37b were predicted. Results: The second structure of IL-37b contained extended strand( 31. 65%),random coil( 52. 75%),alpha helix(8. 26%),beta turn(7. 34%) and the most possible epitopes of IL-37b were located in or adjacent to amino acid 21-27,34-75,175-192,213-215. Conclusion: These results will be helpful for the estimate of the epitopes and provide a theory basis for developing monoclonal antibodies against human IL-37.

Inteleukin-1F7(IL-1 family 7) is a novel member of the IL-1 family.It has been currently named as IL-37.IL-37 has five different subtypes(IL-37a-e).Recent studies found that IL-37b can be combined with α chain of the IL-18 receptor,but does not affect the physiological functions of the IL-18.Binding of IL-37b and IL-18BP(IL-18 binding protein) can enhance the suppression of IL-18BP on IL-18.Some reports showed that IL-37b has significant anti-inflammatory effects.IL-37b plays an important role in a variety of inflammation-related diseases.

Interleukin(IL)-33 is a member of IL-1 family.It is identified as a functional ligand for ST2 which is an IL-1 receptor-like protein.IL-33 / ST2 signaling is involved in T-cell-mediated immune responses.Increasing evidence indicates that IL-33 has different roles in different diseases.Recently,some studies have demonstrated that IL-33 may be related to the genesis and development of fibrosis diseases.We review current knowledge of the biological characteristics of IL-33 and the role of IL-33 / ST2 signaling pathway in fibrosis diseases.

IL-33 is a recently described member of the IL-1 family that has been reported to have a pathogenic role in several inflammatory diseases. In this study, we evaluated the role of IL-33 in a murine model of multiple sclerosis, experimental autoimmune encephalomyelitis (EAE). We showed that the expression of IL-33 and its receptor, ST2, was markedly elevated in the spinal cord of mice during myelin oligodendrocyte glycoprotein (MOG)(35-55) peptide-induced EAE. Administration of a blocking anti-IL-33 antibody in mice of EAE during the induction phase significantly inhibited the onset and severity of EAE and reduced MOG(35-55)-induced IFN-γ and IL-17 production. In contrast, treatment with recombinant IL-33 worsened the disease course of EAE in association with increased induction of both IFN-γ and IL-17. Furthermore, anti-IL-33 treatment caused a remarkable decrease in expression of IL-17, IFN-γ, T-bet and RORγt, and an upregulation of IL-10 and TGF-β in the spinal cord of EAE mice. These results demonstrate that endogenous IL-33 plays a pivotal role in the pathogenesis of EAE and indicate that blockade of IL-33 has a significant protective effect against EAE.

Interleukin-33(IL-33),a novel member of the IL-1 family,is expressed in several tissues.IL-33 binds to the chromosome as a transcriptional repressor.IL-33 also binds to its receptor to induce the activation of T cells,mast cells,basophiles and eosinophils.The receptor of IL-33 is a heterodimeric complex consisting of membrane-bound ST2 and IL-1 receptor accessory protein(IL-RAcP).The IL-33/ST2 signaling pathway plays critical roles in inflammatory and immune diseases,as well as in the functions of central nervous system(CNS).This review will introduce the progress in the research of IL-33,especially the roles of IL-33 in CNS diseases.

2003年,两个独立的研究小组分别发现了一个新的干扰素家族——Ⅲ型干扰素,又被称为干扰素-λ(IFN-λ)[1-2].IFN-λ家族由IFN-λ1、IFN-λ2和IFN-λ3组成,也被分别命名为IL-29、IL-28A和IL-28B.IFN-λ是一个由相关细胞因子组成的称为IL-10-IFN超家族的一部分,这个超家族还包括IL-10相关细胞因子(IL-10、IL-19、IL-20、IL-22、IL-24、IL-26)、Ⅰ型干扰素(13种IFN-α、IFN-3、IFN-ε、IFN-κ、IFN-ω)和Ⅱ型干扰素(IFN-γ)[3].