Striatal delivery of CERE-120, an AAV2 vector encoding human neurturin, enhances activity of the dopaminergic nigrostriatal system in aged monkeys.

Neurturin (NTN) is a potent survival factor for midbrain dopaminergic neurons. CERE-120, an adeno-associated virus type 2 (AAV2) vector encoding human NTN (AAV2-NTN), is currently being developed as a potential therapy for Parkinson's disease. This study examined the bioactivity and safety/tolerability of AAV2-NTN in the aged monkey model of nigrostriatal dopamine insufficiency. Aged rhesus monkeys received unilateral injections of AAV2-NTN into the caudate and putamen, with each animal therefore serving as its own control. Robust expression of NTN within the nigrostriatal system was observed 8 months postadministration. (18)F-fluorodopa imaging using positron emission tomography revealed statistically significant increases in (18)F-fluorodopa uptake in the injected striatum compared with the uninjected side at 4 and 8 months. In addition, at 8 months postadministration, a significant enhancement in tyrosine hydroxylase immunoreactive fibers and an increase in the number of tyrosine hydroxylase immunoreactive cells was observed in the AAV2-NTN injected striatum compared with the uninjected side. Robust activation of phosphorylated extracellular signal-regulated kinase immunoreactivity in the substantia nigra was also observed. Histopathological analyses revealed no adverse effects of AAV2-NTN in the brain. Collectively, these results are consistent with the neurotrophic effects of NTN on the dopaminergic nigrostriatal system and extend the growing body of evidence supporting the concept that AAV2-NTN may have therapeutic benefit for Parkinson's disease.     

HIV GP120对人和无脊椎动物免疫细胞的抑制作用

通过人工合成了人类免疫缺陷性病毒（Ｈｕｍａｎｉｍｍｕｎｏｄｅｆｉｃｉｅｎｃｙｖｉｒｕｓ，ＨＩＶ）糖蛋白肽ＧＰ１２０对人和无脊椎动物（Ｍｙｔｉｌｕｓｅｄｕｌｉｓ）免疫细胞的抑制作用的研究。人单核细胞和Ｍｙｔｉｌｕｓｅｄｕｌｉｓ免疫细胞分别与ＧＰ１２０保温后，均抑制细胞的吞噬细菌（Ｐｓｕｄｏｍｏｎａｓｓｔｒｅｔｚｉ）作用。应用计算机显微图像术（Ｃｏｍｐｕｔｅｒ－ａｓｓｉｓｔｅｄｍｉｃｒｏｓｃｏｐｙ）直     

Inhibition of HIV-1 by modification of a host membrane protease

While it is clear that CD4 Is the receptor for the gp120 envelopeprotein of HIV-1, substantial evidence suggests that other hostcell proteins are required for successful membrane fusion. Studieswere initiated to examine the potential for a protein receptorwhich has an elastase-like character to participate in fusionof HIV-1 with permissive host cells. A synthetic elastase inhibitorwas shown to significantly reduce HIV-1 infectivity when presentduring, but not after, the initial contact between virus andcells. A human T cell elastase-like membrane component was purifiedand shown to be lipid-associated. By competitive Inhibition,the purified protein was shown to bind gp160 within the HIV-1fusion domain. The binding parameters of whole T cell membraneextract, with a hydrophobic pentapeptide representative of thefusion domain, suggested an elastase-like protein is the single,secondary T cell receptor for HIV-1 (K = 1x10³ M^–1). Thepentapeptide interacted with porcine and human (epithelial andpolymorphonuclear leukocyte), but not murine, elastase isoforms,suggesting its participation In the permissiveness of host cellsto infection.     

Comparison of Different Types of Oral Adsorbent Therapy in Patients with Chronic Kidney Disease: A Multicenter,Randomized, Phase IV Clinical Trial

PurposeOral adsorbents delay disease progression and improve uremic symptoms in patients with chronic kidney disease (CKD). DW-7202 is a newly developed oral adsorbent with high adsorptive selectivity for uremic toxins. We evaluated patient preference for and adherence to DW-7202 versus AST-120 therapy and compared treatment efficacy and safety in patients with pre-dialysis CKD.Materials and MethodsA seven-center, randomized, open-label, two-way crossover, active-controlled, phase IV clinical trial was conducted. Patients with stable CKD were randomly assigned to receive DW-7202 (capsule type) or AST-120 (granule type) for 12 weeks. The groups then switched to the other adsorbent and took it for the next 12 weeks. Patient preference was the primary outcome. Secondary outcomes included changes in estimated glomerular filtration rate (eGFR) and serum creatinine, cystatin C, and indoxyl sulfate (IS) levels.ResultsSignificantly more patients preferred DW-7202 than AST-120 (p<0.001). Patient adherence improved after switching from AST-120 to DW-7202; there was no apparent change in adherence after switching from DW-7202 to AST-120. Changes in eGFR and serum creatinine, cystatin C, and IS levels were not significantly different according to adsorbent type. There was also no significant difference in the incidences of adverse events during treatment with DW-7202 and AST-120.ConclusionDW-7202 can be considered as an alternative to AST-120 in patients who cannot tolerate or show poor adherence to granule type adsorbents. Further studies to evaluate factors affecting patient preferences and improved adherence are warranted (Clinical trial registration No. {"type":"clinical-trial","attrs":{"text":"NCT02681952","term_id":"NCT02681952"}}NCT02681952).     

Requirement of the T cell receptor for antigen presentation by T lymphocytes. Effect of envelope glycoproteins of HIV-1 on antigen presentation by T cells.

We have developed CD4+, tetanus antigen-specific T cell clones that proliferate in the presence of tetanus antigen and autologous irradiated peripheral blood leucocytes (PBL) as antigen-presenting cells (APC). There have been several reports that T cells can present antigen themselves. We have used tetanus antigen-specific T cell clones to examine the effects of envelope glycoproteins of HIV-1 on processing and presentation of antigen to T cells. Cloned T cells were pre-incubated with soluble crude preparation of tetanus antigen for 4 h at 37 degrees C, irradiated, and used as APC (T-APC). These cells could present antigen, as assessed by the ability of the autologous cloned T cells to proliferate. Resting T cells and phytohaemagglutinin-activated T cell blasts from autologous PBL could not present tetanus antigen to the responder cloned T cells. Antigen presentation by T-APC was abrogated by treating cells with anti-HLA-DR but not by anti-HLA-DQ monoclonal antibodies; treatment of tetanus antigen-pulsed T-APC with anti-tetanus antibody also blocked the ability of these cells to induce proliferation in responder T cells. Antigen presentation by cloned T cells was by a chloroquine-resistant pathway. Pretreatment of T-APC with envelope glycoprotein of HIV-1, gp120, did not affect the proliferative responses of the responder T cells. These data suggest that gp120 does not inhibit the antigen-presenting function while suppressing antigen-specific responses.     

哇巴因诱导ECV304细胞凋亡与p120ctn mRNA的表达

目的研究哇巴因对ECV304细胞的作用及相关机制。方法应用MTT法检测哇巴因对细胞生长抑制作用,Hoechst33342/PI双荧光染色分析细胞死亡特征,透射电镜和光镜观察细胞形态结构变化,半定量RT-PCR法检测P120ctn 1A和P120ctn 3A mRNA的表达。结果哇巴因以浓度和作用时间依赖的方式抑制ECV304细胞生长;荧光显微镜下可见30%以上细胞为凋亡细胞,20%细胞为晚期凋亡细胞的形态学改变;透射电镜下可见核染色质凝集、边移、凋亡小体形成等细胞形态学变化;RT-PCR结果显示P120ctn 1A mRNA表达较对照组明显降低,P120ctn 3A mRNA表达较对照组明显增强。结论哇巴因能抑制ECV304细胞增殖和诱导其凋亡,并下调P120ctn 1A和上调P120ctn3A mRNA表达。     

Epitope mapping and characterization of a novel CD4-induced human monoclonal antibody capable of neutralizing primary HIV-1 strains

Human immunodeficiency virus (HIV-1) enters target cells by binding its gp120 exterior envelope glycoprotein to CD4 and one of the chemokine receptors, CCR5 or CXCR4. CD4-induced (CD4i) antibodies bind gp120 more efficiently after CD4 binding and block the interaction with the chemokine receptor. Examples of CD4i antibodies are limited, and the prototypes of the CD4i antibodies exhibit only weak neutralizing activity against primary, clinical HIV-1 isolates. Here we report the identification of a novel antibody, E51, that exhibits CD4-induced binding to gp120 and neutralizes primary HIV-1 more efficiently than the prototypic CD4i antibodies. The E51 antibody blocks the interaction of gp120-CD4 complexes with CCR5 and binds to a highly conserved, basic gp120 element composed of the beta 19-strand and surrounding structures. Thus, on primary HIV-1 isolates, this gp120 region, which has been previously implicated in chemokine receptor binding, is accessible to a subset of CD4i antibodies.     

Human immunodeficiency virus gp120 and derived peptides activate protein tyrosine kinase p56Ick in human CD4 T lymphocytes

Human immunodeficiency virus binds to CD4 T lymphocytes by interaction between its envelope glycoprotein gpl20 and the CD4 molecule. The latter is non-covalently associated with a src-related tyrosine kinase, p56^lck. CD4 cross-linking increases the activity of p56^lck, leading to phosphorylation of several cellular substrates. We report here that gpl60/120 increases both the autophos-phorylation of p56^lck and its enzymatic activity (reflected by phosphorylation of an exogeneous substrate) in normal T cells and the HUT78 CD4⁺ T cell line. This effect was detectable 5 min after activation and persisted for 40 min in normal T cells. It did not require gpl20 cross-linking and was associated with phosphorylation of tyrosine residue on several proteins, as shown by phosphotyrosine Western blot analysis. The pattern of proteins phosphorylated on tyrosine residues in response to gpl20 activation was distinct from that induced by anti-CD4 antibodies. p56^lck activation required its association with CD4, since p56^lck activity was not modified in HUT78 T cell lines expressing a truncated or mutated form of CD4 unable to associate with p56^lck. Peptides mimicking residues 418 to 434 and 449 to 464 of HIV-1 Bru gpl20, regions known to participate in gpl20 binding to CD4, also increased p56^lck activity and triggered phosphorylation of similar substrates. Taken together, these results show that gpl60/120 and derived peptides can transiently increase p56^lck activity without the need for CD4 cross-linking. This activation led to a specific pattern of tyrosine phosphorylation on cellular proteins that may be of significance in the biological effects of the gpl20/CD4 interaction, e.g. syncytium formation and inhibition of T cell activation.     

中国株HIV-1 gp120核酸疫苗的实验免疫研究

目的　探讨表达中国株HIV 1gp12 0基因的核酸疫苗在小鼠体内的免疫反应。方法　将表达HIV 1gp12 0的核酸疫苗质粒pVAXP经肌肉注射免疫Balb c小鼠 ,检测免疫小鼠脾CD4 +、CD8+T细胞亚群的数量 ,脾特异性CTL杀伤活性和血清抗体滴度。结果　重组质粒pVAXP免疫组小鼠脾CD4 +、CD8+T细胞亚群的数值均比对照组高 (P <0 .0 5 ) ;免疫组脾特异性CTL杀伤活性与对照组相比差异极显著 (P <0 .0 1) ;血清抗体滴度显著高于对照组 (P <0 .0 5 )。结论　表达HIV 1gp12 0基因的核酸疫苗质粒pVAXP能诱导小鼠产生特异性细胞和体液免疫。     

血栓闭塞性脉管炎患者血栓素、前列环素及血液流变学指标的检测分析

目的 :探讨血栓闭塞性脉管炎患者血栓素B2 、前列环素与血液流变学指标的变化及其与发病机制的关系。方法 :用放射免疫法测定TXA2 和PGI2 的代谢产物血栓素B2 (TXB2 )、6 酮 前列腺素F1α( 6 K PGF1α) ,LBY N6A自清洗旋转式粘度计测定血液流变学指标 ,并与对照组对比分析。结果 :血栓闭塞性脉管炎患者TXB2 ( 53.59± 83.2 1ng/L)、6 K PGF1α( 14 .50± 3.4 5ng/L) ,与对照组相比差异显著 ;全血高切粘度、低切粘度、血浆粘度及纤维蛋白原疾病组均高于对照组 ,差异具有统计学意义。结论 :血栓闭塞性脉管炎患者存在TXA2 /PGI2失衡 ,并与血液流变学的改变有密切关系