Immune systems biology: immunoprofiling of cells and molecules

White blood cells and their secreted products are key elements of immune systems biology that are important indicators of patient health and disease. We have developed the SurroScan microvolume laser scanning cytometer to immunoprofile hundreds of variables, including cell populations, cell surface antigens, and intracellular molecules in antibody-based assays on small samples (about 1 mL) of whole blood, processed blood, or other fluids without cell purification or washing steps. The system enables high-throughput, robust and automated data capture and analysis. We demonstrate the utility of this immunoprofiling technology platform by surveying patient samples before and after glucocorticosteroid administration and show both the expected and novel response characteristics. This system complements recent advances in genomic and proteomic approaches to disease prediction and monitoring.     

Mutation patterns and structural correlates in human immunodeficiency virus type 1 protease following different protease inhibitor treatments

Although many human immunodeficiency virus type 1 (HIV-1)-infected persons are treated with multiple protease inhibitors in combination or in succession, mutation patterns of protease isolates from these persons have not been characterized. We collected and analyzed 2,244 subtype B HIV-1 isolates from 1,919 persons with different protease inhibitor experiences: 1,004 isolates from untreated persons, 637 isolates from persons who received one protease inhibitor, and 603 isolates from persons receiving two or more protease inhibitors. The median number of protease mutations per isolate increased from 4 in untreated persons to 12 in persons who had received four or more protease inhibitors. Mutations at 45 of the 99 amino acid positions in the protease-including 22 not previously associated with drug resistance-were significantly associated with protease inhibitor treatment. Mutations at 17 of the remaining 99 positions were polymorphic but not associated with drug treatment. Pairs and clusters of correlated (covarying) mutations were significantly more likely to occur in treated than in untreated persons: 115 versus 23 pairs and 30 versus 2 clusters, respectively. Of the 115 statistically significant pairs of covarying residues in the treated isolates, 59 were within 8 A of each other-many more than would be expected by chance. In summary, nearly one-half of HIV-1 protease positions are under selective drug pressure, including many residues not previously associated with drug resistance. Structural factors appear to be responsible for the high frequency of covariation among many of the protease residues. The presence of mutational clusters provides insight into the complex mutational patterns required for HIV-1 protease inhibitor resistance.     

A dual-functional paramyxovirus F protein regulatory switch segment: activation and membrane fusion

Many viral fusion-mediating glycoproteins couple alpha-helical bundle formation to membrane merger, but have different methods for fusion activation. To study paramyxovirus-mediated fusion, we mutated the SV5 fusion (F) protein at conserved residues L447 and I449, which are adjacent to heptad repeat (HR) B and bind to a prominent cavity in the HRA trimeric coiled coil in the fusogenic six-helix bundle (6HB) structure. These analyses on residues L447 and I449, both in intact F protein and in 6HB, suggest a metamorphic region around these residues with dual structural roles. Mutation of L447 and I449 to aliphatic residues destabilizes the 6HB structure and attenuates fusion activity. Mutation of L447 and I449 to aromatic residues also destabilizes the 6HB structure despite promoting hyperactive fusion, indicating that 6HB stability alone does not dictate fusogenicity. Thus, residues L447 and I449 adjacent to HRB in paramyxovirus F have distinct roles in fusion activation and 6HB formation, suggesting this region is involved in a conformational switch.     

8-OH-DPAT and MK-801 affect epileptic activity independently of vigilance

Vigilance and parallel occurrence of epileptic activity after administration of the 5-HT_1A agonist 8-OH-DPAT and the NMDA receptor antagonist MK-801 were studied in the genetic absence epilepsy model WAG/Rij rats. Spike-wave discharges (SWD) were present predominantly in passive awake and light slow wave sleep (SWS1) either in control animals or after treatments. Injection of 8-OH-DPAT (20.0 μg/rat i.c.v.) caused marked increase and MK-801 (10.0 μg/rat i.c.v.) decrease in SWD densities, thus the ratios of SWD in passive awake and in SWS1. SWD densities of MK-801 plus 8-OH-DPAT in combination were similar to those of CSF+CSF treated control rats. Both 8-OH-DPAT and MK-801 transiently increased the duration of active awake, increased latency and decreased duration of rapid eye movement (REM) sleep. 8-OH-DPAT increased the amount of SWD despite the decrease in the duration of SWS1. MK-801 decreased the amount of SWD despite the lack of significant change in duration of passive awake or SWS1. Pre-treatment with MK-801 reversed 8-OH-DPAT- induced increase in duration of SWD without any effect on 8-OH-DPAT-induced changes in sleep parameters. Our studies provide evidence that 8-OH-DPAT-induced epileptic activity is independent of its effect on sleep, and that interaction of serotonergic and glutamatergic systems plays a role in the generation of SWD, but not in the regulation of vigilance and sleep.     

昆明地区香石竹病毒病流行状况调查及脱病毒苗的制备

对昆明地区3种不同生产模式下的香石竹（Dianthus caryophyllus L.)进行了调查,采集样本146号,利用酶联免疫法和电镜检测法对样本感染香石竹病毒的情况进行检测,结果表明昆明地区主要流行的香石竹为香石竹斑驳病毒和香石竹坏死斑点病毒,以带香石竹斑驳病毒的香石竹品种“俏新朗”为实验材料,研究了直接剥茎尖法,高温处理结合剥茎尖法和病毒痤处理结合剥茎尖法3种方法在脱病毒效率和茎尖成苗率的差异,实验结果表明以加热处理结合剥茎尖法脱病毒效果最好,0.2mm茎尖脱病毒率可达77．78％,加5％病毒座处理对脱病毒有一定的影响,直接剥茎尖法脱病毒效果最差。     

Trichosanthin inhibits T cell activation by interfering with the recruitment of ZAP-70 to CD3 ζchain

INTRoDUCTIONThichosanthin(Tk),aplantproteinisolatedfromaChinesemedicinalherbTh-1.Correspondenceaddress:Dr.KuangYenCHoU,ShanghaiInstituteofImmunology,Shang-haiSecondMedicalUniversity28oSouthChongqingRoad,Shanghai2ooo25,China.Fax:(8621)63846383,E-mail:my@sh…     

盐胁迫对不同抗盐性小麦叶片荧光诱导动力学的影响

以不同抗盐性小麦为材料,研究了ＮａＣｌ胁迫对叶片叶绿素ａ荧光诱导动力学的影响。指出１５０ｍｍｏｌ／ＬＮａＣｌ使小麦幼苗生长降低,叶绿素含量下降,同时使光系统Ⅱ的潜在光化学活性和原初光能转化率降低,使光反应受到抑制。     

桐花树活性内生真菌筛选及其抗菌化学成分的研究

为了探究桐花树内生真菌在抑菌方面的价值,该文以内生真菌发酵产物的抑菌作用为评价指标筛选活性菌株,采用生物活性跟踪方法结合多种色谱技术分离活性菌株的化学成分,通过波谱与文献数据比对鉴定单体化合物结构,并利用微孔板法测定单体化合物的抑菌活性。结果表明:(1)从桐花树分离得到的16株内生真菌分属2纲7目10科10属,镰刀菌属(Fusarium)为优势菌属。内生真菌GXIMD02029和GXIMD02039的发酵产物对枯草芽孢杆菌、表皮葡萄球菌、耐甲氧西林金黄色葡萄球菌、藤黄微球菌、粘性放线菌和金黄色葡萄球菌有不同程度的抑制作用,GXIMD02038发酵产物对耐甲氧西林金黄色葡萄球菌、藤黄微球菌和金黄色葡萄球菌有抑制作用。(2)7个化合物从内生真菌Phomopsis sp. GXIMD02029中被分离并鉴定为(15R)-acetoxydothiorelone A(1)、cytosporone B(2)、pestalotiopsone H(3)、pestalotiopsone B(4)、4-Hydroxybenzaldehyde(5)、p-Hydroxybenzoic acid(6)、N-(2-phenylethyl)acetamide(7)。(3)化合物1和2有不同程度的抑菌作用,化合物1对枯草芽孢杆菌、表皮葡萄球菌、耐甲氧西林金黄色葡萄球菌的MIC值为16.25 SymbolmA@ g·mL^-1,对藤黄微球菌和粘性放线菌的MIC值为7.812 5 SymbolmA@ g·mL^-1,对金黄色葡萄球菌的MIC值为31.25 SymbolmA@ g·mL^-1。化合物2对藤黄微球菌的MIC值为62.5 SymbolmA@ g·mL^-1,对枯草芽孢杆菌、表皮葡萄球菌、耐甲氧西林金黄色葡萄球菌、粘性放线菌的MIC值为125 SymbolmA@ g·mL^-1,对金黄色葡萄球菌的MIC值为250 SymbolmA@ g·mL^-1。该文筛选了3株活性菌株,首次报道化合物1具有抗菌活性,为桐花树内生真菌在抗菌价值方面提供了依据。