Characterisation of a 34 kD protein from potato cyst nematodes, using monoclonal antibodies with potential for species diagnosis

Two monoclonal antibodies, which differentially recognise the two species of potato cyst nematodes (PCN), Globodera pallida and G. rostochiensis, are described. They have been shown to have potential for quantification of these two species, recognising proteins of the same molecular weight (34 kD) in both species. Further investigation showed these proteins to have isoelectric points at pH values of 5.7 in G. pallida and 5.9 in G. rostochiensis, in common with the proteins used by Fleming & Marks (1983) to differentiate the species of PCN. They are likely to be structurally very similar, with the same physiological function (and therefore similar concentrations) in the two species. In cross-reactivity tests with a wide range of soil nematode species, the antibodies reacted strongly only with species of the genus Globodera, and thereby confirmed their potential as the basis of a quantitative immunoassay likely to be useful in management of PCN populations.     

诱导受体3（DcR3）在胆管癌中的表达及意义

目的:观察胆管癌组织及血清中诱导受体3(decoy receptor 3,DcR3)蛋白的表达及其临床价值。方法:采用免疫组化S-P法检测45例胆管癌、15例癌旁胆管正常组织中DcR3蛋白的表达,ELISA法检测31例胆管癌及18例胆道良性疾病患者和28例正常人外周血清中DcR3的水平。结果:45例胆管癌组织中DcR3阳性表达29例,阳性率为64.4%,胆管正常组织中无阳性表达。DcR3的表达与肿瘤临床分期、肿瘤浸润和转移有关(P<0.05)。胆管癌患者及胆管良性疾病患者血清DcR3水平分别为152.2535.94 pg/ml,98.35 14.27 pg/ml,均高于正常人。胆管癌患者与胆道良性疾病患者血清DcR3水平相比差异有显著性(P<0.01)。结论:DcR3在胆管癌组织中表达增高。DcR3的表达与胆管癌的发生、发展以及转移有关,可成为治疗胆管癌的一个新靶点。血清DcR3的检测对胆管癌的诊断有一定的临床价值。     

Spiggin levels are reduced in male sticklebacks infected with Schistocephalus solidus

Using an enzyme-linked immunosorbent assay (ELISA) based technique, Schistocephalus solidus infection was shown to considerably reduce levels of spiggin in the kidney of male three-spined sticklebacks Gasterosteus aculeatus from an oligotrophic upland lake. These results suggested a graduated effect of the infection on the reproductive physiology of male three-spined sticklebacks.     

猪和牛轮状病毒抗原和抗体的检测

应用ELISA直接双抗体夹心法检查轮状病毒抗原,24份仔猪和29份犊牛的腹泻粪样,分别有12和16份阳性。用病毒RNA电泳分析检查阳性粪样,各出现两种病毒RNA电泳型,用中和试验检查17份成年牛和16份成年猪血清,分别有16和15份病毒抗体阳性。将其与ELISA间接法和结合法进行了比较。     

蛇毒类凝血酶鼠单克隆抗体的制备

采用杂交瘤技术,获得了４株稳定分泌抗蛇毒类凝血酶的单克隆抗体杂交瘤细胞株,均属ＩｇＧ１ｋ链,４株杂交瘤细胞培养上清液效价为 ４ × １０－１～４ × １０－２,腹水效价为 ４ × １０－１～３．２ ×１０－５。     

猴T淋巴细胞趋向性病毒1型双抗原夹心ELISA检测试剂盒的研制

目的研制灵敏度和特异性高的检测实验猴血清中T淋巴细胞趋向性病毒-1型（STLV-1型）／E体的双抗原夹心ELISA（dsELISA）检测试剂盒。方法采用经原核表达系统表达并纯化的人T淋巴细胞白血病病毒-1型（HTLV-1型）的Env蛋白作为包被用抗原,建立了检测STLV-1的dsELISA诊断方法。通过优化反应条件和筛选试剂,确定了dsELISA诊断试剂盒的相关条件,并经敏感性、特异性和重复性试验考查该试剂盒质量。结果试剂盒特异性好,批内重复试验变异系数（CV）〈7％,批间重复试验CV〈10％。对200份猴血清进行随机检测,与国际公认的诊断试剂盒（美国BioReliance公司）的符合率为97％。结论本试剂盒可初步应用于临床上实验猴STLV-1型抗体的检测。     

Barnase-barstar high affinity interaction phenomenon as the base for the heterogenous bioluminescence pseudorabies virus' immunoassay

The effective new variant of "sandwich" bioluminescent enzyme immunoassay (BEIA) for the sensitive detection of glycoprotein B (gB) of pseudorabies virus (PrV) was presently developed. The high affinity interaction of barnase-barstar protein pair and photoprotein obelin as bioluminescent marker were for the first time successfully applied to BEIA development. Preliminary the two monoclonal antibodies, 11/5 and 34/2, were raised against gB for ELISA PrV detection. Presently we used the same immuno-"sandwich" principle for BEIA. To do this the two different bioconjugates were elaborated. Recombinant barnase was chemically conjugated with monoclonal anti-PrV's gB IgG, and also barstar was fused in frame to obelin. The characteristics of BEIA method have been compared to ELISA PrV detection. We have shown the proposed here gB-BEIA was 40-fold more sensitive as opposed to gB-ELISA test. The construction might have a broad promise in multiple potential immunological applications.     

HLA class II alleles and measles virus-specific cytokine immune response following two doses of measles vaccine

Measles virus-specific T cells and the production of cytokines play a critical role in the immune response following measles immunization. To understand the genetic factors that influence variation in IFN- and IL-4 responses following measles immunization and to provide insight into the factors influencing both cellular and humoral immunity to measles, we assessed associations between human leukocyte antigen (HLA) class II genes and measles-specific Th1 and Th2-type cytokine responses in peripheral blood lymphocytes from 339 children previously vaccinated with two doses of measles-mumps-rubella vaccine (MMR-II). Median values for measles-specific IFN- and IL-4 secretion levels were 40.73 and 9.71 pg/ml, respectively. The global tests suggested associations between measles-specific IFN- response and alleles of the DRB1 and DQB1 loci (P=0.07 and P=0.02, respectively). Specifically, DRB1*0301, *0901, and *1501 alleles were significantly associated with IFN- secretion. The alleles that suggested evidence of an HLA association with IL-4 secretion were DRB1*0103, *0701, and *1101. Th1 cytokine responses and DQB1 allele associations revealed that the alleles with the strongest association with IFN- secretion were DQB1*0201, *0303, *0402, and *0602. Specific alleles with a suggestive association with low measles-specific Th2 cytokine responses were DQB1*0202 and *0503. In addition, DPB1*0101, *0201, and *0601 alleles provided suggestive evidence of an HLA association with measles-induced IFN- response, while DPB1*0501 was associated with an IL-4 response. These data suggest that IFN- and IL-4 cytokine responses to measles may be genetically restricted in part by HLA class II genes, which in turn can restrict the cellular immune response to measles vaccine.     

Dolichos yellow mosaic virus belongs to a distinct lineage of Old World begomoviruses; its biological and molecular properties

Dolichos yellow mosaic disease (DYMD) affects the production of dolichos in South Asia. Diseased plants produce characteristic bright yellow mosaic patches on the leaves and early infections cause reductions in yield. The putative dolichos yellow mosaic virus (DoYMV) was transmitted poorly (maximum 18.3% transmission) by the whitefly, Bemisia tabaci. DoYMV has a narrow host range and infected only Lablab purpureus and L. purpureus var. typicum out of the 36 species tested. Virus was detected using monoclonal antibodies in a triple‐antibody sandwich enzyme‐linked immunosorbent assay and by PCR. Complete DNA‐A components of DoYMV isolates from Mysore and Bangalore, South India, were sequenced, but several attempts to identify DNA‐B and DNA‐β were unsuccessful. DoYMV isolates shared DNA‐A nucleotide identities of 92.5–95.3% with previously described isolates from North India and Bangladesh. They were most similar to mungbean‐infecting begomoviruses at 61.6–64.4% of DNA‐A nucleotide identities. Phylogenetic analyses of DNA‐A sequences grouped the dolichos‐infecting and mungbean‐infecting begomoviruses into a distinct cluster away from begomoviruses infecting non‐leguminous plants in the Indian subcontinent. Antigenically, legume‐infecting begomoviruses were most similar to each other compared with non‐legume viruses. Collectively, these results indicate that legume‐infecting begomoviruses in the Indian subcontinent belonged to a distinct lineage of Old World begomoviruses.