黄鳝IgM的分离纯化及兔抗黄鳝IgM抗血清的制备

目的分离纯化黄鳝血清免疫球蛋白,制备其兔抗血清,并检测抗血清的特异性。方法用Protein A亲和层析的方法纯化黄鳝血清免疫球蛋白,通过SDS-聚丙烯酰胺凝胶电泳检测蛋白的纯度,免疫大耳白兔制备抗血清,利用免疫双扩散检测抗血清的效价,通过western blotting检测抗血清的特异性。结果纯化了黄鳝血清免疫球蛋白,免疫双扩散法测定兔抗黄鳝免疫球蛋白血清效价为1∶32,western blotting结果显示抗血清具有很好的特异性。结论成功纯化了黄鳝免疫球蛋白,制备了兔抗黄鳝IgM抗血清,为建立黄鳝的血清学检测系统奠定了基础。     

内皮素在肠源性内毒素血症发病中的作用

本实验在整体大鼠和离体大鼠回肠血管灌流模型上,观察外源性给予内皮素和内皮素抗血清对肠道内毒素转运的影响。结果表明,内皮素能促进健康大鼠肠道内毒素的转运,内皮素抗血清能拮抗循环中的内毒素促进肠道内毒素转运的作用。提示,内皮素可能是循环内毒素促进肠道内毒素转运的中间介质之一,参与肠源性内毒素血症的发病过程。     

Specificity enhancement of polyclonal antisera by the induction of tolerance to unwanted cross‐reacting determinants

Steroids form a structurally closely related group. As a result, antibodies produced for use in immunoassays regularly show unwanted cross‐reactivities. These may be reduced by altering hapten‐protein coupling procedures, thereby reducing the exposure of the determinants giving rise to the undesirable cross‐reaction. However, these procedures can prove to be complex, expensive and not totally predictable in outcome. Exploitation of the clonal selection theory is an attractive alternative approach. The host is primed with the interfering cross‐reactant coupled to a non‐immunogenic amino acid copolymer to inactivate the B‐lymphocyte clones specific for this steroid, producing a specific immunotolerance. Then, 3 days later, the host is immunized with the steroid against which an antibody is required. The clones producing antibody to this immunogen are unaffected and the cross‐reactivity is significantly reduced or deleted. The technique has been applied to the reduction of endogenous sex steroid cross‐reactivity from antibodies prepared against synthetic and semi‐synthetic androgens (17α‐methyltestosterone, 19‐nor‐ß‐testosterone) and the progestogen medroxyprogesterone. Antibodies prepared against the synthetic oestrogen zeranol using this technique have significantly reduced its undesirable cross‐reactivity with the fungal metabolite 7α‐zearalenol. Highly specific antisera have been generated in all cases, the only adverse effect being a reduction in the titres achieved in comparison with rabbits receiving the conventional immunizing regime.     

华支睾吸虫成虫RPEF基因产物及其抗RPEF多克隆抗体的制备

目的体外制备华支睾吸虫成虫RNA聚合酶Ⅱ延长因子（RPEF）基因产物及抗RPEF多克隆抗体。方法亲和纯化试剂盒纯化表达蛋白pGEx4T-1,RPEF,凝血酶酶切表达蛋白制备重组蛋白RPEF,将重组蛋白RPEF免疫小鼠,制备多克隆抗血清,检测抗体滴度和抗血清特异性。结果体外制备的RPEF蛋白经SDS电泳呈单一条带;酶联免疫吸附结果显示,免疫过程抗体滴度呈连续上升趋势;免疫印迹结果显示,小鼠抗血清具有抗RPEF特异性。结论获得较专一的华支睾吸虫RPEF蛋白和RPEF抗血清。     

Simple Serological Assays for Detecting Rice Tungro Viruses

An attempt was made to produce sensitive and specific polyclonal antisera against the viruses causing rice tungro disease, and to assess their potential for use in simple diagnostic tests. Using a multiple, sequential injection procedure, seven batches of polyclonal antisera against rice tungro bacilliform virus (RTBV) and rice tungro spherical virus (RTSV) were produced. These were characterized for their sensitivity and specificity using ring-interface precipitin test and double antibody sandwich (DAS) ELISA. Thirty-one weeks after the first immunization, antiserum batch B6b for RTBV showed the highest ring interface titer (DEP = 1:1920). For RTSV, batches S3, S4b and S5b all had similar titres (DEP = 1:640). In DAS-ELISA, however, significant differences among purified antisera (IgG) batches were observed only at IgG dilution of 10^-3. At that dilution, IgGB4b showed the greatest sensitivity, while IgGS3 showed greatest sensitivity for RTSV. When all IgG batches were tested against 11 tungro field isolates (dual RTBV-RTSV infections) at sample dilution of 1:10, IgGB4b and IgGB6b for RTBV and IgGS3 and IgGS6b for RTSV performed equally well. However, after cross adsorption with healthy plant extracts in a specially prepared healthy plant-Sepharose affinity column, only IgGB6b could be used specifically to detect RTBV in a simple tissue-print assay.     

Quantitative Immunoelectrophoretic Study of Genus Pityrosporum Sabouraud

Summary: The antigens of the three species of genus Pityrosporum were compared by quantitative immunoelectrophoresis (QIE). Antigen-antibody reference systems were established for Pityrosporum orbiculare, Pityrosporum pachydermatis and two morphologically different strains (A und F) isolated in the laboratory from Pityriasis versicolor lesions. Crossed Immunoelectrophoresis (CIE) demonstrated 63,47,50 and 35 precipitation peaks, respectively. This provides evidence of the complex antigenic structure of these species, which had never been demonstrated. In those systems, the immunologic comparison by two QIE methods of the proteins of three strains of Pityrosporum orbiculure, one strain of Pityrosporium ovale and two strains A and F, did not establish differences. The two strains of Pityrosporum pachydermatis were found to be markedly different from the other six, as nearly 40% of the proteins studied were immunologically different. We, therefore, consider, as several authors do, that Pityrosporum ovale and Pityrosporum orbiculare are actually synonyms. So, the genus Pityrosporum consists of two species: one is lipid-dependent, Pityrosporum ovale, and the other is not, Pityrosporum pachydermatis. Zusammenfassung: Die Antigene der drei Artender Gattung Pityrosporum wurden mit Hilfe der quantitativen Immunelektrophorese (QIE) verglichen. Für Pityrosporum orbiculare, Pityrosporum pachydermatis und zwei morphologisch unterschiedliche Stänune (A und F), die in unserem Laboratorium aus Pityriasis versicolor-Herden isoliert worden waren, wurden Antigen-Antikörper-Vergleichssysteme aufgestellt. In der gekreuzten Immunelektrophorese (CIE) konnten 63, 47,50 und 35 Fällungsgipfel (peaks) aufgezeigt werden. Damit konnte eine bisher nicht nachgewiesene Komplexität der Antigenstrukturen dieser Spezies demonstriert werden. In den genannten Systemen waren keine Unterschiede beim immunologischen Vergleich mit Hilfe von zwei QIE-Methoden der Proteine von drei Stämmen von Pityrosporum orbiculare, einem Stamm von Pityrosporum ovale und den zwei Stämmen A und F festzustellen. Die beiden Stämme von Pityrosporum pachydermatis unterschieden sich deutlich von den übrigen sechs Stämmen, da fast 40% der untersuchten Proteine immunologisch abwichen. Wir sind daher mit verschiedenen anderen Autoren der Meinung, daß Pityrosporum ovale und Pityrosporum orbiculare Synonyme sind. Daher besteht die Gattung Pityrosporum aus zwei Arten: dem lipidabhängigen Pityrosporum ovale und dem nicht lipidabhängigen Pityrosporum pachydermatis.     

抗猴IgM和IgG免疫血清的制备研究

本文用Sephadex G200柱层析及DEAE-Sephadex A50离子交换层析法,从正常猴血清中提取猴IgM及IgG,经SDS-PAGE证实为电泳纯。2Kg左右的家兔,腹股沟皮下注射BCG后2周,在肿大的淋巴结内及基周围注射与福氏完全佐剂(FCA)混合的猴IgM或IgG;并在间隔40天及20天再用与福氏不完全佐剂(FIA)混合的猴IgM或IgG加强免疫。用琼脂糖双扩散法、免疫电泳、2ME还原及SPA吸收后的抗体捕捉ELISA等方法证实,所得兔抗猴免疫血清效价高,特异性强,为研究恒河猴的体液免疫应答提供了必不可少的材料。     

78份细胞毒抗血清的HLA-A.B位点特异性鉴定

本文用微量淋巴细胞毒方法鉴定出28份血清,作为分型试剂的使用单价、双价特异性血清。前后已鉴定出作为分型试剂血清共62份,含18种特异性血清,用这些分型试剂检出沈阳地区汉族HLA—A位点抗原的基因频率为0.756,仍有空白0.244,HLA—B位点抗原的基因频率为0.7780,空白0.2220。     

Immunocytochemistry of human brain tissue with a polyclonal antiserum against neuropeptide Y

Summary NPY-containing neuronal structures in the cereoral cortex of surgical tissue samples were compared to those in postmortem material by immunocytochemical methods. However, the quality of preservation of individual neurons and axonal and dendritic plexuses in the neuropil is unusually fine in the surgical specimens. This result is most likely attributable to the excellent fixation that can be regularly achieved by rapid and careful handling of tissue during and after surgical removal. The tissue is suitable for both light and electron microscopy, and the superior preservation also leads to intense, reliable antibody reactions. Postmortem tissue samples can provide good specimens for immunocytochemistry when properly handled as previously described. However the postmortem delays prior to fixation disrupt neuronal integrity in the immunostained structures. Nevertheless, postmortem material from carefully studied subjects of neurological diseases compared with age matched controls can provide valuable information.     

Acute lymphoblastic leukaemia (ALL) associated antigen—I. Expression in different haematopoietic malignancies

The cellular specificity of antisera to non-T, non-B ALL has been assessed in a series of 545 patients (adults and children) either presenting with untreated acute or chronic leukaemia or in relapse. The ‘ALL associated antigen’ was detected and evaluated by indirect immunofluorescence using standard microscopy and with the Fluorescence Activated Cell Sorter. Anti-ALL serum reacts with cells from a variable proportion of patients in the following groups: non-T, non-B ALL; Ph¹ positive acute leukaemia or CML in blast crisis; AUL; and a few lymphomas. A weak but definite expression of the ALL antigen was also detected in 10% of ALLs with a thymic phenotype (Thy-ALL). Cross-absorbtion studies indicate that a single antigen or antigenic complex is shared by these various cell types. These observations are interpreted to suggest that several clinically and karyotypically distinct leukaemias involve a similar lymphoid or pre-lymphoid cell type and that the ALL associated antigen is most likely a normal gene product of that cell type or lineage involved in the disease.