裙带菜和萱藻凝集素对刺参组织主要免疫酶活性的影响

研究刺参基础饲料中添加不同剂量裙带菜凝集素和萱藻凝集素对刺参酸性磷酸酶(ACP)、碱性磷酸酶(AKP)、溶菌酶(LSZ)、过氧化氢酶(CAT)活性的影响。在基础饲料中分别添加0.58%、1.16%、2.32%裙带菜凝集素(质量分数);0.32%、0.64%、1.28%萱藻凝集素,以基础饲料饲组为对照,于投喂后第4、8、13、17天检测ACP、AKP、LSZ、CAT活性。结果显示,投喂17 d内,各实验组ACP活性随时间和剂量增加持续升高,ACP活性均高于对照组;LSZ活性则随时间延长持续升高,但萱藻凝集素各组活性在第17天有所下降,且LSZ活性与凝集素添加量成正比关系;裙带菜凝集素组CAT活性呈升高趋势,萱藻凝集素组呈先高后低规律变化。     

Variation in Relative Amounts of Isolectins in Bread Wheat (Triticum aestivum) Landraces from Northern Nigeria and Chad

Wheat germ agglutin (WGA) present in the embryo of a bread wheat grain is a mixture of three isolectins, viz CL_A, CL_B and CL_D which are coded by genes located on die chromosomes 1A, 1B and 1D. Their respective absolute amounts were determined in 62 Nigerian and five Chad landraces. Added was the Madagascar variety ‘Kanem’ which is stated to be selected from Chad material. Some of the Nigerian landraces were formerly identified as derived from Indian Pakistani wheat varieties/landraces introduced into Northern Nigeria before 1940. These had relatively high CL_D/GL_A and CL_B/CL_A ratios. Other Nigerian landraces with such high ratios were postulated to come from India/Pakistan, too. The Chad wheats, except ‘Kanem’, have WGA amounts and-ratios similar to those native to Northern Nigeria. This may point to a common history. ‘Kanem’ is said to be bred in Madagascar from Chad wheats. Apparently it has a different origin. So the determination of the WGA amount and its isolectin composition is a helpful means to identify the history of a land race or of an improved variety.     

Adjuvant and immunostimulatory effects of a D-galactose-binding lectin from Synadenium carinatum latex (ScLL) in the mouse model of vaccination against neosporosis

Vaccination is an important control measure for neosporosis that is caused by a coccidian parasite, Neospora caninum, leading to abortion and reproductive disorders in cattle and serious economic impacts worldwide. A D-galactose-binding lectin from Synadenium carinatum latex (ScLL) was recently described by our group with potential immunostimulatory and adjuvant effects in the leishmaniasis model. In this study, we evaluated the adjuvant effect of ScLL in immunization of mice against neosporosis. First, we investigated in vitro cytokine production by dendritic cells stimulated with Neospora lysate antigen (NLA), ScLL or both. Each treatment induced TNF-α, IL-6, IL-10 and IL-12 production in a dose-dependent manner, with synergistic effect of NLA plus ScLL. Next, four groups of C57BL/6 mice were immunized with NLA + ScLL, NLA, ScLL or PBS. The kinetics of antibody response showed a predominance of IgG and IgG1 for NLA + ScLL group, whereas IgG2a response was similar between NLA + ScLL and NLA groups. Ex vivo cytokine production by mouse spleen cells showed the highest IFN-γ/IL-10 ratio in the presence of NLA stimulation for mice immunized with NLA + ScLL and the lowest for those immunized with ScLL alone. After parasite challenge, mice immunized with NLA + ScLL or ScLL alone presented higher survival rates (70-80%) and lower brain parasite burden as compared to PBS group, but with no significant changes in morbidity and inflammation scores. In conclusion, ScLL combined with NLA was able to change the cytokine profile induced by the antigen or lectin alone for a Th1-biased immune response, resulting in high protection of mice challenged with the parasite, but with low degree of inflammation. Both features may be important to prevent congenital neosporosis, since protection and low inflammatory response are necessary events to guide towards a successful pregnancy.     

海带和条斑紫菜凝集素提取液对刺参体壁主要免疫酶活性的影响

为探讨凝集素提取液对刺参Apostichopus japonicus免疫的影响,将刺参基础饲料中分别添加质量分数为0·64%、1·28%、2·56%的海带凝集素提取液,或质量分数为1·88%、3·76%、7·52%的条斑紫菜凝集素提取液,分别投喂6个试验组刺参(体质量为3·50 g ±1·95 g),以基础饲料饲养组为对照,分别于投喂凝集素提取液后第4、8、12、17天时,检测各组刺参体壁中过氧化氢酶( CAT)、溶菌酶( LSZ)、碱性磷酸酶(AKP)和酸性磷酸酶(ACP)的活性。结果表明：投喂海带凝集素提取液后,各浓度组刺参CAT、 LSZ活性在第8、12、17天时均有显著或极显著提高(P<0·05或P<0·01),0·64%组ACP活性在第8、17天时有显著提高(P<0·05);投喂条斑紫菜凝集素提取液后,各浓度组刺参CAT活性在第8、12、17天时有显著或极显著提高( P<0·05或P<0·01),7·52%组LSZ活性在第8天时有显著提高( P<0·05), AKP活性在第4、17天时有极显著提高( P<0·01), ACP活性在第12天时7·52%组有极显著提高( P<0·01),第17天时各浓度组均有显著或极显著提高(P<0·05或P<0·01)。研究表明,海带和条斑紫菜凝集素提取液均能提高剌参体壁中CAT、 LSZ、 AKP、 ACP的活性,显著增强剌参非特异性免疫水平,将其作为刺参免疫增强剂具有广阔的应用前景。     

鱼类凝集素概述

鱼类凝集素成员种类繁多、功能多样,是鱼体免疫系统的重要组成部分。随着生物技术的发展,人们不断从鱼体内分离出多种具有生物活性的凝集素,对鱼类凝集素的研究逐年增多。以鱼类凝集素为研究对象,结合国内外研究成果,对鱼类凝集素的分类及功能等进行阐述,以期为今后的研究及应用提供一定的参考。     

Establishment of TaqMan Real-time Quantitative PCR Assay for Foreign Gene Copy Numbers in Transgenic Soybean

TaqMan quantitative PCR technique was used to detect the copies of exogenous CaMV35S flanks sequence in transgenic soybean. With soybean lectin as the endogenous reference gene, and gene complex DNA in...     

菲律宾蛤仔凝集素的分离纯化及抑菌活性研究

采用Cellulose DE52离子交换层析、Sephadex G- 100分子筛层析及TSK G3000PWXL凝胶色谱柱HPLC从菲律宾蛤仔Ruditapes philippinarum中分离纯化出具有N-乙酰半乳糖酰胺(N- acetyl- D- galactosamine,GaINAc)和猪胃黏蛋白(Mucin from porcine stomach,PSM)特异性的菲律宾蛤仔凝集素(简称为MCL-T).SDS-PAGE分析结果表明,MCL-T的相对分子量为148000,含有62000和36000的亚基.抑菌试验结果表明,MCL-T对金黄色葡萄球菌Staphylococcus aureus和枯草芽孢杆菌Bacillus subtilis有较高的抑菌活性.对MCL-T中Trp残基、-S-S-、-SH以及Tyr残基进行化学修饰后,MCL-T对金黄色葡萄球菌的抑菌活性丧失;对其-S-S-进行化学修饰后,MCL-T对枯草芽孢杆菌的抑菌活性影响不大,而对其-SH、Trp残基和Tyr残基进行化学修饰后,MCL-T对枯草芽孢杆菌的抑菌活性丧失.研究表明,MCL-T 肽链上的氨基酸残基状态与其抑菌活性有关.     

血凝法测定饲料中植物凝集素含量

通过研究植物凝集素(lectin)的血细胞凝集活性与其浓度的关系，提出了用血凝法测定饲料中lectin的新方法，并探讨了试样粉碎粒度、脱脂方法、脱脂时间、抽提时间、抽提次数等测定条件对测定结果的影响。结果表明，lectin的活力与浓度之间存在高度线性相关(R^2=0．99)。血凝法测定饲料中lectin的含量时，以样品粉碎过40目、样品与30～60℃石油醚或乙醚比例(W／V)1：8脱脂8h、样品与生理盐水(W／V)1：8抽提8h为宜。在上述条件下测定，回收率可达88％～102％。经过对8种不同的饲料及原料样品lectin含量的测定，平均相对偏差为0．9％～2．4％，变异系数为1．3％～3．3％。结果表明，利用血凝法可快速测定饲料中的lectin含量。     

Comparison of the amino acid sequences of acorn barnacle lectins showing different inhibitory activities toward the crystal growth of calcium carbonate

ABSTRACT: The amino acid sequence of a D -galactose-binding lectin isolated from the hemolymph of the acorn barnacle Balanus rostratus was determined. The lectin (BRL) ( M _r 120K) is a multimeric protein whose subunit consists of 182 amino acids. The amino acid sequence was compared with those of multiple lectins (BRA-2, BRA-3) from Megabalanus rosa to explore the relationship between the structures and the inhibitory activity toward the crystal growth of calcium carbonate. Although BRL was 46% identical to BRA-2 and 15% identical to BRA-3, the lectin had no inhibitory activity, unlike BRA-2 and BRA-3. Both the number and the localization of acidic amino acid residues and their amide forms were different among them. Observations by scanning electron microscopy revealed modifications in the size and morphology of the calcium carbonate crystals grown in the presence of the lectins.