Monoclonal antibodies inhibit prion replication and delay the development of prion disease

Prion diseases such as Creutzfeldt-Jakob disease (CJD) are fatal, neuro-degenerative disorders with no known therapy. A proportion of the UK population has been exposed to a bovine spongiform encephalopathy-like prion strain and are at risk of developing variant CJD. A hallmark of prion disease is the transformation of normal cellular prion protein (PrP(C)) into an infectious disease-associated isoform, PrP(Sc). Recent in vitro studies indicate that anti-PrP monoclonal antibodies with little or no affinity for PrP(Sc) can prevent the incorporation of PrP(C) into propagating prions. We therefore investigated in a murine scrapie model whether anti-PrP monoclonal antibodies show similar inhibitory effects on prion replication in vivo. We found that peripheral PrP(Sc) levels and prion infectivity were markedly reduced, even when the antibodies were first administered at the point of near maximal accumulation of PrP(Sc) in the spleen. Furthermore, animals in which the treatment was continued remained healthy for over 300 days after equivalent untreated animals had succumbed to the disease. These findings indicate that immunotherapeutic strategies for human prion diseases are worth pursuing.     

Correlation of structural elements and infectivity of the HET-s prion

Prions are believed to be infectious, self-propagating polymers of otherwise soluble, host-encoded proteins. This concept is now strongly supported by the recent findings that amyloid fibrils of recombinant prion proteins from yeast, Podospora anserina and mammals can induce prion phenotypes in the corresponding hosts. However, the structural basis of prion infectivity remains largely elusive because acquisition of atomic resolution structural properties of amyloid fibrils represents a largely unsolved technical challenge. HET-s, the prion protein of P. anserina, contains a carboxy-terminal prion domain comprising residues 218-289. Amyloid fibrils of HET-s(218-289) are necessary and sufficient for the induction and propagation of prion infectivity. Here, we have used fluorescence studies, quenched hydrogen exchange NMR and solid-state NMR to determine the sequence-specific positions of amyloid fibril secondary structure elements of HET-s(218-289). This approach revealed four beta-strands constituted by two pseudo-repeat sequences, each forming a beta-strand-turn-beta-strand motif. By using a structure-based mutagenesis approach, we show that this conformation is the functional and infectious entity of the HET-s prion. These results correlate distinct structural elements with prion infectivity.     

A 'unified theory' of prion propagation

There is now very persuasive evidence that the transmissible agent for spongiform encephalopathies such as scrapie, consists of a modified form of the normal host protein PrPc, devoid of any nucleic acid. On the other hand, because there are many different strains of scrapie agent with distinct phenotypes which can be propagated in animals homozygous for the PrPc gene, it has been suggested that a nucleic acid must be a component of the agent. Can the two views be reconciled?     

2例透明细胞软骨肉瘤的误诊分析

目的通过对误诊病例分析,探讨透明细胞软骨肉瘤影像学特征及其鉴别诊断方法。方法对两例透明细胞软骨肉瘤临床、影像及病理资料作系统整理并分析。结果两例肿瘤均位于长骨骨端,影像学似良性肿瘤表现,早期准确的影像学诊断非常困难,需影像、临床及病理密切结合。鉴别诊断结果为:浸润性骨巨细胞瘤与动脉瘤样骨囊肿。结论透明细胞软骨肉瘤影像学趋向良性肿瘤表现,生物学行为却为恶性,需采用广泛性切除手术方式治疗,因此早期、正确的影像学诊断至关重要。     

Antibodies against analogous heptad repeat peptide HR212 of Newcastle Disease Virus inhibit virus-cell membrane fusion

Membrane fusion is a key step in enveloped virus entry. Highly conserved heptad repeat regions (HR1 and HR2) of Newcastle disease virus (NDV) fusion protein (F) are critical functional domains for viral membrane fusion. They display different conformations in the membrane fusion states and are viewed as candidate targets for neutralizing antibody responses. We previously reported that an analog of heptad repeat peptides HR2-HR1-HR2(HR212) and HR2 could inhibit NDV induced cell-cell membrane fusion. Here, we show that HR212 can induce the production of highly potent antibody in immunized rabbits, which could recognize full length peptides of both HR1 and HR2, and inhibit NDV hemagglutination and NDV entry. These suggest that either HR212 or its antibody could be an inhibitor of virus-induced cell-cell membrane fusion.     

Antibodies against analogous heptad repeat peptide HR212 of Newcastle Disease Virus inhibit virus-cell membrane fusion

Membrane fusion is a key step in enveloped virus entry. Highly conserved heptad repeat regions (HR1 and HR2) of Newcastle disease virus (NDV) fusion protein (F) are critical functional domains for viral membrane fusion. They display different conformations in the membrane fusion states and are viewed as candidate targets for neutralizing antibody responses. We previously reported that an analog of heptad repeat peptides HR2-HR1-HR2(HR212) and HR2 could inhibit NDV induced cell-cell membrane fusion. Here, we show that HR212 can induce the production of highly potent antibody in immunized rabbits, which could recognize full length peptides of both HR1 and HR2, and inhibit NDV hemagglutination and NDV entry. These suggest that either HR212 or its antibody could be an inhibitor of virus-induced cell-cell membrane fusion.     

植物细胞培养实验在细胞工程实验教学中的应用

为提高细胞工程实验教学的效果,培养学生的实验操作能力和解决问题能力,探索了适合于实验教学的具体化和可操作的植物细胞培养方案.大叶黄杨(Euonymus japonicus)和紫花地丁(Viola philippica)可作为植物细胞培养的理想实验材料;诱导愈伤组织的最适培养基为基本培养基+2,4-二氯苯氧乙酸2 mg/L+6-苄氨基腺嘌呤0.4 mg/L.     

岩黄连离体细胞培养及生物碱成分分析

选取岩黄连不同组织部位作为外植体诱导愈伤组织，调整基本培养基和初始pH值，经多次继代培养，获得稳定生长的岩黄连离体细胞；采用薄层层析（TLC）、紫外扫描（UV）和高效液相色谱（HPLC）等方法对细胞和植株中生物碱成分进行分析测定。结果表明，叶片最适于诱导愈伤组织；最佳基本培养基为B5，初始pH值为6．0；岩黄连离体培养细胞的生长周期为20d左右，第15天生物量最高，达到15．3g／L（DW）；细胞和植株中生物碱成分有较大差别，但都舍有脱氢卡维丁，细胞中脱氢卡维丁的含量为0．58mg／g。