Structural analysis of human and macaque mAbs 2909 and 2.5B: implications for the configuration of the quaternary neutralizing epitope of HIV-1 gp120

The quaternary neutralizing epitope (QNE) of HIV-1 gp120 is preferentially expressed on the trimeric envelope spikes of intact HIV virions, and QNE-specific monoclonal antibodies (mAbs) potently neutralize HIV-1. Here, we present the crystal structures of the Fabs of human mAb 2909 and macaque mAb 2.5B. Both mAbs have long beta hairpin CDR H3 regions >20 ? in length that are each situated at the center of their respective antigen-binding sites. Computational analysis showed that the paratopes include the whole CDR H3, while additional CDR residues form shallow binding pockets. Structural modeling suggests a way to understand the configuration of QNEs and the antigen-antibody interaction for QNE mAbs. Our data will be useful in designing immunogens that may elicit potent neutralizing QNE Abs.     

万寿菊属不同品种初花期抗旱特性分析

以万寿菊属(Tagetes)9个品种为试验材料,研究了自然持续干旱胁迫对它们初花期的花最大直径、叶色、旱害指数等形态指标以及叶绿素含量(Chl a+b,Ch a/b)、叶片相对含水量(RWC)、叶片保水力(WHC)、叶片和花的脯氨酸、可溶性糖和可溶性蛋白含量等生理指标的影响,以揭示其抗旱特性及其生理机制.结果显示:(1)持续6d干旱胁迫条件下,万寿菊9个品种花的最大直径显著降低,叶绿素含量和相对含水量均呈明显下降趋势.(2)万寿菊叶片的脯氨酸、可溶性蛋白和可溶性糖含量均呈上升趋势;而花的脯氨酸含量持续上升,可溶性蛋白含量呈下降趋势,但可溶性糖含量变化趋势复杂.(3)万寿菊旱害指数与其叶片相对含水量、叶绿素总含量、叶片和花的脯氨酸含量、叶片可溶性蛋白含量呈极显著相关.研究表明,抗旱性强的品种可以通过调节自身的渗透调节物质含量减轻干旱伤害;9个品种初花期抗旱性强弱依次为:珍妮>金门>鸿运>拳王>巨人>发现>小英雄>大英雄>迪阿哥.     

浮床生态场空间分布特征

在原位试验的基础上,建立浮床生态场模型.模拟结果表明,浮床生态势值在"场源"处最大,从浮床"场源"向周围水平方向以及向水下垂直方向,随着距离的增加生态势逐渐下降.同时,浮床植物的生物量对生态场效应有显著影响,浮床植物生长越旺盛、生物量越大,生态场效应越强.1 m×1m的浮床,当植物生物量为39.40 kg·m-2时,浮床系统可最远影响到距离浮床1.86 m范围内的水体,并对浮床周围10.90 m2范围内的水体产生作用.该模型可以模拟浮床生态场的空间分布特征及其随植物生长发育的动态变化过程,并可通过模型定量化反映浮床系统在其作用范围内对任意一点的作用强度及其综合影响,该模型的构建将为生态浮床的野外应用提供理论依据.     

狂犬病病毒糖蛋白重组人腺病毒的构建及免疫效果的初步研究

构建表达狂犬病病毒弱毒SRV9糖蛋白(GP)的重组人5型腺病毒,检测其对小鼠的免疫效果。将狂犬病病毒SRV9株GP基因的完整开放阅读框克隆到腺病毒表达系统中的穿梭质粒多克隆位点,构建重组穿梭质粒pac-Ad5CMV-Gs9,以罗氏转染液介导线性化骨架质粒和重组穿梭质粒共转染293AD细胞,细胞病变后取培养物进行PCR鉴定并电镜观察,在293AD细胞上测定病毒滴度。以106 TCID50重组腺病毒腹腔接种昆明小鼠,免疫后不同时段采尾静脉血通过荧光抗体病毒中和试验(FAVN)检测小鼠血清狂犬病中和抗体效价。正确构建重组穿梭质粒pacAd5CMV-Gs9;获得表达狂犬病病毒SRV9株GP蛋白的缺陷型重组人5型腺病毒;病毒滴度达到106 CFU/mL以上;腹腔接种小鼠14d后均产生了抗狂犬病中和抗体,有效保护率达90%。成功获得了表达狂犬病病毒GP基因的重组腺病毒,该腺病毒免疫小鼠可产生保护性中和抗体,为进一步开发新型兽用狂犬病疫苗奠定了物质基础。     

海拔梯度对巴郎山奇花柳叶片δ13C的影响

2010年在四川卧龙自然保护区选择海拔为2350、2700、3150和3530 m的4个分布地点,研究了巴郎山海拔梯度对奇花柳叶片13C、光合、CO2扩散导度、氮含量、光合氮利用效率( PNUE)和比叶面积(SLA)的影响.结果表明:随着海拔的升高,目标树种叶片氮含量(尤其是单位面积氮含量)及PNUE增加,叶片δ13C值也随之显著增加,且海拔每升高1000 m,δ13C增加1.4‰;CO2扩散导度(气孔导度和叶肉细胞导度)的增加,在一定程度上阻碍了叶片δ13C值随海拔升高,但不足以改变δ13C值随海拔升高的趋势;羧化能力是羧化位点与外界CO2分压比( Pc/Pa),甚至δ13C的限制因子.在海拔2350～2700m,奇花柳光合系统内部氮素分配主要受温度的影响,而2700～3530 m的光照作用可能更大.奇花柳的SLA随海拔无显著变化.     

微生物菌剂在农业废弃物堆肥腐熟过程中的应用及其田间试验效果

以农村生活垃圾中的可堆肥腐熟成分和蘑菇渣为堆肥原料,通过添加微生物菌剂进行堆肥试验,研究其在农业废弃物堆肥腐熟过程中的作用,并通过田间试验研究堆肥腐熟后肥料样品对黄瓜和青椒的增产效果,以验证其肥效。结果表明添加微生物菌剂有助于堆肥腐熟后样品的氮、磷、钾的保全和有机质的增加,促进养分均衡。添加微生物菌剂的堆肥腐熟肥料样品在田间试验中对黄瓜和青椒的增产效果最为显著,分别为22.21%和19.87%。     

IL-15:IL-15 receptor alpha superagonist complex: high-level co-expression in recombinant mammalian cells, purification and characterization

IL-15, a promising cytokine for treating cancer and viral diseases, is presented in trans by the IL-15 receptor (IL-15R) alpha-chain to the IL-15Rβγc complex displayed on the surface of T cells and natural killer (NK) cells. We previously reported that an asparagine to aspartic acid substitution at amino acid 72 (N72D) of IL-15 provides a 4-5-fold increase in biological activity compared to the native molecule. In this report, we describe Chinese hamster ovary (CHO) cell expression of a soluble complex (IL-15 N72D:IL-15RαSu/Fc) consisting of the IL-15 N72D superagonist and a dimeric IL-15Rα sushi domain-IgG1 Fc fusion protein. A simple but readily scalable affinity and ion exchange chromatography method was developed to highly purify the complex having both IL-15 binding sites fully occupied. The immunostimulatory effects of this complex were confirmed using cell proliferation assays. Treatment of mice with a single intravenous dose of IL-15N72D:IL-15RαSu/Fc resulted in a significant increase in CD8+ T cells and NK cells that was not observed following IL-15 treatment. Pharmacokinetic analysis indicated that the complex has a 25-h half-life in mice which is considerably longer than <40-min half-life of IL-15. Thus, the enhanced activity of the IL-15N72D:IL-15RαSu/Fc complex is likely the result of the increased binding activity of IL-15N72D to IL-15Rβγc, optimized cytokine trans-presentation by the IL-15RαSu domain, the dimeric nature of the cytokine domain and its increased in vivo half-life compared to IL-15. These findings indicate that this IL-15 superagonist complex could serve as a superior immunostimulatory therapeutic agent.     

Artificial gravity with ergometric exercise preserves the cardiac, but not cerebrovascular, functions during 4 days of head-down bed rest

Cardiovascular and musculoskeletal deconditioning occurring in long-term spaceflight requires new strategies to counteract these adverse effects. We previously reported that a short-arm centrifuge produced artificial gravity (AG), together with ergometer, has an approving effect on promoting cardiovascular function. The current study sought to investigate whether the cardiac and cerebrovascular functions were maintained and improved using a strategy of AG combined with exercise training on cardiovascular function during 4-day head-down bed rest (HDBR). Twelve healthy male subjects were assigned to a control group (CONT, n=6) and an AG combined with ergometric exercise training group (CM, n=6). Simultaneously, cardiac pumping and systolic functions, cerebral blood flow were measured before, during, and after HDBR. The results showed that AG combined with ergometric exercise caused an increase trend of number of tolerance, however, there was no significant difference between the two groups. After 4-day HDBR in the CONT group, heart rate increased significantly (59±6 vs 66±7 beats/min), while stroke volume (98±12 vs 68±13 mL) and cardiac output (6±1 vs 4±1 L/min) decreased significantly (p<0.05). All subjects had similar drops on cerebral vascular function. Volume regulating hormone aldosterone increased in both groups (by 119.9% in CONT group and 112.8% in the CM group), but only in the CONT group there were a significant changes (p<0.05). Angiotensin II was significantly increased by 140.5% after 4-day HDBR in the CONT group (p<0.05), while no significant changes were observed in the CM group. These results indicated that artificial gravity with ergometric exercise successfully eliminated changes induced by simulated weightlessness in heart rate, volume regulating hormones, and cardiac pumping function and partially maintained cardiac systolic function. Hence, a daily 1h alternating +1.0 and +2.0 Gz with 40 W exercise training appear to be an effective countermeasure against cardiac deconditioning.