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61.
Isolated Light Chain of Tetanus Toxin Inhibits Exocytosis: Studies in Digitonin-Permeabilized Cells 总被引:3,自引:2,他引:1
Previous work indicates that the heavy chain of tetanus toxin is responsible for the binding of the toxin to the neuronal membrane and its subsequent internalization. In the present study, the light chain of tetanus toxin mimicked the holotoxin in inhibiting Ca2+-dependent secretion of [3H]norepinephrine from digitonin-permeabilized adrenal chromaffin cells. Preincubation of tetanus toxin with monoclonal antibodies to the light chain prevented the inhibition by tetanus toxin. Preincubation of tetanus toxin with nonimmune ascites fluid or with monoclonal antibodies directed against the C fragment (the C-terminal of the heavy chain) or the heavy-chain portion of the B fragment did not prevent inhibition by tetanus toxin. The data indicate that the light chain is responsible for the intracellular blockade of exocytosis. 相似文献
62.
A subclone of rat pheochromocytoma cells expresses high affinity receptors for tetanus toxin on differentiation with NGF [Walton, K.M., Sandberg, K., Rogers, T.B. and Schnaar, R.L. (1988) J. Biol. Chem. 263, 2055–2063]. In the presence of protein cross-linking agents, [125I]tetanus toxin, bound to these cells at 0°C, forms a cross-linked product with apparent molecular weight of 120 kDa. The formation of [125I]tetanus toxin conjugate involves the heavy chain of the toxin, is prevented by cold toxin and it is largely reduced by pretreating cells with proteases, The cross-linked product is formed only upon incubation of the toxin with NGF-differentiated cells. These results suggest that a protein with apparent molecular weight of 20 kDa is involved in the neurospecific binding of tetanus toxin. 相似文献
63.
《Biologicals》2014,42(3):160-168
Meningococcal group X (MenX) is responsible for recent outbreaks of meningitis reported in sub-Saharan region of Africa. Although protective vaccines are available for meningitis, they are not effective against MenX. An efficacious, monovalent conjugate vaccine was designed against MenX and a fed-batch fermentation process was developed. The MenX polysaccharide (PS) was purified and yield estimated to be 15-fold higher than the reported elsewhere. Structure of MenX polysaccharide was confirmed by 1H, 13C NMR spectroscopy analysis. Molecular weight of PS was found to be 310 kDa using HPLC-SEC coupled to refractive index (RI) detector. The MenX–Tetanus toxoid (TT) monovalent conjugate proved to be highly immunogenic in mice, and the bactericidal titers of MenX–TT conjugate were 10-fold higher than native PS. Increasing the dose of MenX–TT conjugate from 0.5 μg to 1.0 μg induced an 8-fold higher antibody titer as well as serum bactericidal titer. The current work suggests that the MenX–TT conjugate is a candidate vaccine against meningitis caused by Meningococcal group X strains. 相似文献
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65.
目的:实现副溶血性弧菌溶血毒素基因tdh克隆与表达,并以表达产物TDH作免疫原,研究其对海水鱼免疫活性的影响.方法:PCR扩增tdh;构建重组质粒(pET-28-TDH);IPTG诱导表达;SDS-PAGE检测:以纯化的TDH融合蛋白脱毒为类毒素免疫原免疫健康真鲷,测定其血清中过氧化氢酶(CAT)、酸性磷酸酶(ACP)、超氧化物歧化酶(SOD)和碱性磷酸酶(AKP)活性变化;用副溶血性弧菌病原攻毒,检测免疫保护作用.结果:tdh基因成功克隆并表达;表达蛋白相对分子量为21 kDa,证实为TDH蛋白;类毒素免疫真鲷后,其血清中总超氧化物歧化酶(T- SOD)、酸性磷酸酶(ACP)、碱性磷酸酶(AKP)和过氧化氢酶(CAT)活力在注射后48h、24h、24h、2Ah达到最高,最高点分别高于对照组77%、328%、75%、381%;72 h后均恢复至对照组水平.结论:类毒素对真鲷免疫系统具有明显的刺激作用;类毒素免疫后对攻毒的真鲷保护率达50%. 相似文献
66.
三种免疫制剂对真鲷弧菌病的免疫保护性 总被引:9,自引:3,他引:6
菌体疫苗按不同的方式对真鲷进行免疫2周后,对实验鱼均具有免疫保护性,免疫保护性最好的免疫组,免疫保护率在初次免疫后高达60%,强化免疫后免疫保护率可提高到80%;粗制LPS经去毒处理后初次免疫真鲷,不同浓度的LPS对实验鱼具有不同程度的免疫保护性,强化免疫后,免疫保护率均有明显的提高,浓度越高,免疫保护性越强,对真鲷的免疫保护率最高可达90%,最小弧菌产生的外毒素经福尔马林灭活后制成毒素苗,这种毒素苗能产生较好的免疫保护性,其免疫保护率可达80%,这表明外毒素不仅是最小弧菌产生的毒力因子,同时也是菌体产生的有效保护性抗原。 相似文献
67.
A Chimeric protein of CFA/I,CS6 subunits and LTB/STa toxoid protects immunized mice against enterotoxigenic Escherichia coli
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Narges Zeinalzadeh Ali Hatef Salmanian Goli Goujani Jafar Amani Ghasem Ahangari Asal Akhavian Mahyat Jafari 《Microbiology and immunology》2017,61(7):272-279
68.
Andrey I. Tchorbanov Jordan D. Dimitrov Tchavdar L. Vassilev 《World journal of microbiology & biotechnology》2004,20(2):211-217
The use of semi-synthetic broths for cultivation of Corynebacterium diphtheriae instead of a meat extract-based broth avoids the presence of highly undesirable bovine meat antigens in the diphtheria toxoid. As information on the composition of casein digest-based broths used for the production of diphtheria toxoid is scarce, we have now developed one. The composition of a casein-based medium that supports vigorous bacterial growth as well as high toxin production is described below. The comparative analysis of the toxoids, produced using the meat-based Pope–Lingood and the casein digest-based broths, showed considerable differences in their molecular composition. The variance of weight distribution of toxoid-containing molecular complexes was smaller when the semi-synthetic broth was used. Normal human therapeutic IgG recognizes some of the proteins in the meat-based medium but does not react with any components of the semi-synthetic medium. While precipitation at the isoelectric point of the diphtheria toxoid produced by culturing the C. diphtheriae strain in the semi-synthetic medium resulted in a preparation meeting the requirement for purity (more than 1500 limit floculation Lf/mg protein nitrogen PN), the toxoid produced in the Pope–Lingood broth failed to meet this requirement in some cases, even after a second purification step using ultrafiltration. 相似文献
69.
It has been previously shown that 5-HT uptake inhibition produced by tetanus toxin (TeTx) corresponds to a non-competitive inhibition, and it is preceded by phosphorylation of the tyrosine-kinase receptor trkA, phospholipase C activation and translocation of protein kinase C isoforms [FEBS Lett. 481 (2000) 177; FEBS Lett. 486 (2000) 136]. In the present work, it is shown that agonists of tyrosine-kinase receptors (NGF, EGF, basic FGF) enhance Na+-dependent, 5-hydroxytryptamine (serotonin, 5-HT) uptake in the synaptosomal-enriched P2 fraction from rat-brain, suggesting a divergence in the intracellular signal pathways triggered by TeTx and by agonists of TyrK receptors. Co-applications of TeTx and agonists of TyrK receptors result in a mutual and partial reversion of their effects on 5-HT transport. In spite of their differences on transport, TeTx, TPA and NGF produce an increase in serotonin transporter phosphorylation in Ser separately, which is abolished by the PKC-inhibitor bisindolylmaleimide-1. Co-application of sodium vanadate, a tyrosine-phosphatase inhibitor, partially abolishes the effect produced by TeTx, whereas genistein, a tyrosine-kinase inhibitor, does not exert any variation of TeTx inhibition. Analyses by immunoblotting of the activation of specific PKC isoforms activation, determined as translocation to the membrane compartment, reveals differences in the pattern produced by NGF and TeTx. PKCγ, δ, and isoforms are equally activated by both compounds, whereas the β isoform is activated in a sustained manner only by TeTx, and the isoform is only down-regulated by NGF. The aim of the present work was to explore whether NGF have the same effect on 5-HT transport than TeTx, since both compounds share the ability of activate part of the same transduction pathways. In spite of this, growth factors and TeTx show an opposite effect on 5-HT transport, even though SERT phosphorylation is enhanced in both cases. The differential effect on - and β-PKC isoenzymes found between NGF and TeTx action could explain this apparent discrepancy. 相似文献
70.