Expression of lymphotactin mRNA in experimental crescentic glomerulonephritis

Lymphotactin (LTN) is the sole member of C chemokine, the third subfamily of chemokines. LTN has been shown to be a chemoattractant specific for CD8⁺ cells and/or natural killer (NK) cells, and to be produced by CD8⁺ T cells, NK cells, and mast cells. However, there have been no reports describing its expression in clinical or experimental models of diseases so far. Since glomerular infiltration of CD8⁺ cells is prominent in an animal model of crescentic glomerulonephritis induced in WKY rats by an injection of anti-glomerular basement membrane antibody, we investigated the gene expression of LTN in this model. LTN mRNA was not detected in normal glomeruli but was detected at 0.5 h after the antibody injection, which detection preceded the infiltration of CD8⁺ cells. The expression of LTN mRNA peaked on day 3 and decreased thereafter. We next studied the expression of LTN mRNA in cultured glomerular and vascular cells, and found that glomerular mesangial and vascular endothelial cells could express LTN mRNA when stimulated with IL-1β. These results indicate that the gene expression of LTN is enhanced in the animal model of glomerulonephritis and that intrinsic renal cells are the potential source of the gene expression of LTN in the kidney.     

单核细胞源性树突状细胞表达淋巴细胞趋化因子mRNA的初步研究

目的:体外诱导、培养单核细胞源性树突状细胞(DC),研究其淋巴细胞趋化因子(lymphotactin,Lptn)mRNA表达的动态变化。方法:采用密度梯度离心的方法分离人外周血中的单个核细胞(PBMC),用重组人粒细胞-巨噬细胞集落群体刺激因子(rhGM-CSF)、重组人白细胞介素-4(rhIL-4)刺激贴壁的单核细胞,诱导培养DC,第6天用重组人肿瘤坏死因子-α(rhTNF-α)诱导DC成熟。用流式细胞术检测成熟和未成熟的DC表面分子CD1a和CD83;在电镜下观察成熟DC的形态;以RT-PCR法扩增其LptncDNA并克隆至pGM-TEasyT载体中,测序;以RT-PCR结合凝胶成像分析系统,半定量分析培养3、5及7dDC的LptnmRNA表达的强度。结果:电镜观察培养7d的细胞具有典型的DC形态,流式细胞术检测DC表面分子CD83呈高水平表达。用RT-PCR法克隆的cDNA序列与GenBank中U23772(登陆号)提供的序列一致。培养3d的DC不表达LptnmRNA,培养7d的DC较培养5d的DCLptnmRNA表达增强。结论:单核细胞源性DC能表达LptnmRNA,随着DC的成熟,LptnmRNA的表达增强。     

肾移植后并发肺结核感染的诊断与治疗

背景：肾移植后患者肺结核感染率较高,临床表现缺乏典型性,给诊断和治疗带来不便。 目的：总结同种异体肾移植后肺结核感染的诊断和治疗方法。 方法：回顾性分析2010年1月至2013年10月期间在南方医科大学珠江医院器官移植科诊断为肾移植后肺结核感染的13例患者相关临床诊断和治疗方法。 结果与结论：肾移植后并发肺结核感染的患者发病时间为肾移植后4-120个月,62%(8/13)患者于移植后18个月内发病。患者多以长时间发热为主要的临床表现,常以低热为首发表现。4例根据病史、影像学资料结合病原学阳性确诊,5例根据病史、影像学资料结合肺穿刺活检组织病理学阳性确诊,其余4例根据病史、影像学资料结合实验性抗结核治疗有效而做出临床诊断。患者肺部体征早期不明显,胸部CT有助于早期诊断和鉴别诊断。所有患者遵循早期、规律、全程、适量、联合原则进行抗结核治疗,疗程一般6-10个月,经给予联合抗结核感染药物、调整免疫抑制剂及五酯胶囊保肝等综合治疗,13例患者均存活,未出现死亡病例。2例由于感染早期未及时正规治疗,发生急性排斥反应,导致移植肾功能丧失而恢复血液透析,其余患者均痊愈出院,随访6个月肾功能正常(查血肌酐变化)。 说明肾移植后并发肺结核病的患者应早发现、早诊断、早治疗。CT引导下穿刺活检可作为肾移植后菌阴肺结核诊断和鉴别的有效且可行的手段。在调整免疫方案和抗结核治疗同时给予五酯胶囊可显著减少钙调神经蛋白抑制剂类药物剂量,减轻钙调神经蛋白抑制剂类药不良反应。 中国组织工程研究杂志出版内容重点：肾移植;肝移植;移植;心脏移植;组织移植;皮肤移植;皮瓣移植;血管移植;器官移植;组织工程全文链接：     

树突状细胞在结核病免疫应答中的作用及机制

作为最有效的专职抗原提呈细胞（DC）,树突状细胞在结核病免疫中的作用日益受到关注。机体感染结核杆菌后,未成熟DC（iDC）捕获抗原并逐渐发育成熟,携带抗原成份从感染组织迁移至外周免疫器官,将抗原成份提呈给T、B淋巴细胞激发免疫应答,起着连接固有免疫和适应性免疫作用,并通过分泌IL-12、IFN-γ等细胞因子参与机体免疫调节。深入研究DC在结核病免疫中的作用机制将为抗结核新型疫苗的开发及免疫治疗方案的设计提供依据。     

Interleukin 17-Producing γδ T Cells Increased in Patients with Active Pulmonary Tuberculosis

Although it has been known that γδ T cells may play an important role in the immune response to infection of Mycobacterium tuberculosis （M. tb）, the mechanisms by which the γδ T cells participate in the innate and/or acquired immunity to tuberculosis （TB） have not been full elucidated. In the present study, 27 patients with active pulmonary TB and 16 healthy donors （HD） were performed. We found that proportion of IL-17-producing cells among lymphocyte was similar between TB patients and HD, whereas the proportions of γδ T cells in IL-17-producing cells （59.2%） and IL-17-producing cells in γδ T cells （19.4%） in peripheral blood were markedly increased in TB patients when compared to those in HD （43.9% and 7.7%, respectively）. In addition, the proportions of IFN-T-producing γδ T cells in TB patients were obviously lower than that in HD. Upon re-stimulated with M. tb heat-treated antigen （M. tb-HAg） in vitro, fewer IL-17-producing γδ T cells were generated from HD and TB patients, whereas IFN-T-producing γδ T cells were increased in TB patients compared to that in HD. Our findings in TB patients and healthy human were consistent with other murine investigation that the IL-17- producing γδ T cells were main source of IL-17 in mouse model of BCG infection, suggesting that γδ T cells might be involved in the formation of tubercular granuloma in pulmonary TB patients, but need further identification.     

结核分枝杆菌Rv0073基因在大肠杆菌中的表达及纯化

目的 构建结核分枝杆菌(MTB) Rv0073基因原核表达载体并进行表达和纯化.方法 以MTB H37Rv基因组DNA为模板,采用聚合酶链反应(PCR)扩增目的基因片段,构建原核表达载体pET26b-Rv0073,经测序确定无误后转化至大肠杆菌(E.coli)感受态细胞BL21中.用聚丙烯酰氨凝胶电泳(SDS-PAGE)方法检测重组蛋白表达,检测异丙基-β-D-硫代半乳糖苷(IPTG)诱导不同时间、不同温度条件下重组蛋白表达量.采用His镍磁珠进行外源蛋白小量纯化.结果 成功构建重组表达质粒,重组蛋白经IPTG诱导后,2h开始明显表达且表达量无时间依赖性,在不同温度诱导下,重组蛋白的表达量随温度的增高而减少.重组蛋白以包涵体形式存在,经His镍磁珠纯化后获得重组蛋白.结论 成功构建并表达Rv0073蛋白,为后续Rv0073的大量纯化及其功能研究奠定了基础.     

Expression,purification and characterization of Mycobacterium tuberculosis RpfE protein

Resuscitation promoting factor E(RpfE)is one of the five Rpf-like proteins in Mycobacterium tuberculosis(M.tuberculosis).These Rpf-like proteins are secretory,which make them candidates for recognition by the host immune system.In this study,the RpfE gene was amplified from M.tuberculosis,cloned into the expression vectors pDE22 and pPRO EXHT,and were expressed in Mycobacterium vaccae(M.vaccae)and Escherichia coli DH5α,respectively.Both recombinant RpfE proteins were purified by Ni-Sepharose affinity chromatography,and were given to C57BL/6 mice.The RpfE proteins elicited T cell proliferation,and stimulated the production of gamma interferon(IFN-γ),interleukin-10(IL-10)and IL-12.Our results indicated that the RpfE protein expressed in M.vaccae could more efficiently stimulate cellular immune response,making it a promising candidate as a subunit vaccine.