Soluble HLA/peptide monomers cross-linked with co-stimulatory antibodies onto a streptavidin core molecule efficiently stimulate antigen-specific T cell responses

Soluble MHC–peptide complexes, commonly referred to as tetramers, have been shown to induce strong cross-linking of TCR and CD8, resulting in a vigorous activation followed by a rapid non-apoptotic CD8⁺ T cell death. This has limited tetramer use for antigen-specific T cells isolation and cloning, as sorted tetramer positive cells were shown to possess compromised functional integrity. Here we show that the cross-linking of a secondary co-stimulatory signal into oligomeric MHC:peptide complexes prevents such cell death, and in contrast strongly stimulates antigen-specific T cell responses. Such soluble antigen-presenting complexes (sAPCs) containing MHC:peptide complexes linked to either anti-CD27 or anti-CD28 antibodies were capable of priming and expanding HLA-A*0201 restricted CMV specific T cells and also of generating functional HLA-A*0301 restricted BCR/ABL-specific T cell responses. These sAPCs constitute an encouraging alternative method for generating antigen-specific T cells that could be applied to a variety of antigens. A. I. Dodi and P. J. Travers contributed equally to this work. This paper is an original contribution from the meeting which took place on 28 and 29 May 2008 in Nottingham, UK, celebrating the contribution of Prof. I. A. “Tony” Dodi (^†29.1.2008) to the EU project “Network for the identification and validation of antigens and biomarkers in cancer and their application in clinical tumour immunology (ENACT)”.     

Transgenic tobacco plants expressing Tarin 1 inhibit the growth of Pseudomonas syringae pv. tomato and the development of Spodoptera frugiperda

The protein Tarin 1, from Colocasia esculenta, was expressed in Nicotiana tabacum. Bioassays were done on plants expressing Tarin 1 at different levels using Spodoptera frugiperda larvae, various bacteria and fungi and the root‐knot nematode Meloidogyne javanica. It was found that S. frugiperda larvae fed on transformed plants had retarded and lower pupation, lower accumulated biomass and higher mortality rate than larvae fed on control plants. Also, Tarin 1 was found to inhibit the growth in vitro of Pseudomonas syringae pv. tomato. For Meloidogyne javanica, both relative replication and root damage were greater in control plants than in transformed plants, but the results were not statistically significant. This work illustrates the effects of plants expressing Tarin 1, on the growth and development of insects and bacteria, and shows its potential for pest management.     

High frequency of homozygosity of the HLA region in melanoma cell lines reveals a pattern compatible with extensive loss of heterozygosity

Malignant transformation of cells is frequently associated with abnormalities in human leukocyte antigen (HLA) expression. MHC class I loss or down-regulation in cancer cells is a major immune escape route used by a large variety of human tumours to evade antitumour immune responses mediated by cytotoxic T lymphocytes. The goal of our study was to explore HLA genotyping and phenotyping in a variety of melanoma tumour cell lines. A total of 91 melanoma cell lines were characterised for HLA class I and II genotype. In addition, 61 out of the 91 cell lines were also analysed for HLA class I and II cell surface molecule expression by flow cytometry. Unexpectedly, we found that 19.7% of the melanoma cell lines were homozygous for HLA class I genotypes, sometimes associated with HLA class II homozygosity (8.79%) and sometimes not (10.98%). The frequency of homozygosity was significantly higher compared with the control groups (1.6%). To identify the reasons underlying the high frequency of HLA homozygosity we searched for genomic deletions using eight pairs of highly polymorphic microsatellite markers covering the entire extended HLA complex on the short arm of chromosome 6. Our results were compatible with hemizygous deletions and suggest that loss of heterozygosity on chromosome arm 6p is a common feature in melanoma cell lines. In fact, although autologous normal DNA from the patients was not available and could not be tested, the retention in some cases of heterozygosity for a number of microsatellite markers would indicate a hemizygous deletion. In the rest of the cases, markers at 6p and 6q showed a single allele pattern indicating the probable loss of part or the whole of chromosome 6. These results led us to conclude that loss of heterozygosity in chromosome 6 is nonrandom and is possibly an immunologically relevant event in human malignant melanoma. Other well-established altered HLA class I phenotypes were also detected by flow cytometry that correspond to HLA class I total loss and HLA-ABC and/or specific HLA-B locus down-regulation.     

Sorghum Insect Problems and Management

Sorghum (Sorghum bicolor) has high levels of starch,sugar,and fiber and is one of the most important energy crops in the world.Insect damage is one of the challenges that impacts sorghum biomass production.There are at least 150 insect species that can infest sorghum varieties worldwide.These insects can complete several generations within a growing season,they target various parts of sorghum plants at developmental stages,and they cause significant biomass losses.Genetic research has revealed the existence...     

桃儿七光合生理特性的地理差异研究

对中国由南向北5个种源地桃儿七(Sinopodophyllum hexandrum)的光合生理生态特性进行了研究.结果表明:(1)北部宁夏六盘山地区植株的光饱和点(LSP)和光补偿点(LCP)最高,表观光量子效率(AQY)、瞬时光能利用效率(ILUE)和最大光合速率(Pmax)最低,光合能力最差;最南部云南纳帕海植株的LSP和LCP很低,但因其AQY和ILUE最高,故其Pmax最大,强光下的光合能力最高.(2)最南部云南纳帕海和最北部甘肃兴隆山个体的羧化效率(CE)最高,且CO2补偿点(CCP)较低,因此CO2利用率较高;而较南部四川刷经寺个体的CE最低,且CCP最高,故CO2利用效率最低.(3)各种源植株叶片的叶绿素总量及叶绿素a含量无显著差异,而不同地区个体的Chla/Chlb值差异显著,最北部甘肃兴隆山植株的Chla/Chlb值最大,而南部四川刷经寺的Chla/Chlb值最小.(4)各种源地植株的水分利用效率(WUE)和蒸腾速率(Tr)对光量子通量密度(PFD)与CO2浓度的响应过程也表现出显著的地理分异.总的来看,桃儿七是一种喜光但又较耐阴植物,但不同分布区桃儿七的光合生理生态特征已经出现了较显著的地理分化.     

A型肉毒毒素轻链基因的克隆及其结构分析

以献报道的A型肉毒毒素基因全序列为标准,设计并合成一对引物,自肉毒梭菌基因组中扩增出肉毒毒素轻链基因片段,并将扩增产物与pGEM—T载体在体外连接,构建测序重组质粒,进行测序和基因结构分析。PCR扩增获得了产物为1 364bp的DNA片段,测序结果与DNA数据库对照检索分析证明,此基因片段与GenBank中的A型肉毒毒素LC基因的一致性达99．9％以上,可以认为克隆的基因为A型肉毒毒素LC基因。     

卤虫染色体倍性组成的研究

本文报道了我国4个地理品系卤虫的染色体倍性组成及卵径分布,分析了卤虫染色体的非整倍性,讨论了卤虫种的分布。 天津卤虫有2倍体(2n=42),占33.3％和5倍体,占16.2％;染色体数的波动为21—108;卵径为262±17μm。 海南卤虫有2倍体(2n=42),占8％;4倍体,占17.7％;5倍体,占18.3％;染色体数的波动为16—111;卵径为285±15μm。 新疆卤虫有2倍体(2n=42),占39.2％;染色体数的波动为17—106;卵径为269±15μm。 山西卤虫有2倍体(2n=42),占45.1％,染色体数的波动为17—84;卵径为234±13μm。