Carcinoembryonales Antigen (CEA) und CEA-like Aktivität in Ascites und Pleuraergüssen

Zusammenfassung Das carcinoembryonale Antigen (CEA) wurde in 18 nicht-malignen und 21 malignen Ergüssen bestimmt. In 95% der benignen Ergüsse lag es im Normbereich und war lediglich in einem Fall bei Klebsiellen-Pneumonie erhöht. Von den 21 malignen Ergüssen war das CEA in 57% der Fälle erhöht und in 4 Fällen war der erhöhte Titer erster Hinweis auf die Malignität des Ergusses. Teilweise bestehen erhebliche Unterschiede in der Titerhöhe zwischen Erguß und Serum. Die gleichzeitige Bestimmung des carcinoembryonalen Antigens in Erguß und Serum erhöht die diagnostische Aussagekraft.     

T cell receptor alpha variable 12‐2 bias in the immunodominant response to Yellow fever virus

The repertoire of human αβ T‐cell receptors (TCRs) is generated via somatic recombination of germline gene segments. Despite this enormous variation, certain epitopes can be immunodominant, associated with high frequencies of antigen‐specific T cells and/or exhibit bias toward a TCR gene segment. Here, we studied the TCR repertoire of the HLA‐A*0201‐restricted epitope LLWNGPMAV (hereafter, A2/LLW) from Yellow Fever virus, which generates an immunodominant CD8⁺ T cell response to the highly effective YF‐17D vaccine. We discover that these A2/LLW‐specific CD8⁺ T cells are highly biased for the TCR α chain TRAV12‐2. This bias is already present in A2/LLW‐specific naïve T cells before vaccination with YF‐17D. Using CD8⁺ T cell clones, we show that TRAV12‐2 does not confer a functional advantage on a per cell basis. Molecular modeling indicated that the germline‐encoded complementarity determining region (CDR) 1α loop of TRAV12‐2 critically contributes to A2/LLW binding, in contrast to the conventional dominant dependence on somatically rearranged CDR3 loops. This germline component of antigen recognition may explain the unusually high precursor frequency, prevalence and immunodominance of T‐cell responses specific for the A2/LLW epitope.     

去甲肾上腺素影响单核细胞抗原提呈的机制

１０＾－１３～１０＾－９ｍｏｌ／Ｌ的去甲肾上腺素体外作用时能促进人外周血单核细胞的抗原提呈功能（ＡＰＦ）。ＰＫＣ激活剂ＰＭＡ和抑制剂４ａ－ＰＤＤ分别能加强和抑制ＮＥ对Ｍｏｎ的ＡＰＦ的促进作用；异搏定能抑制ＮＥ的这种作用，而ＰＫＡ的抑制剂ＰＫＩ对ＮＥ的这种作用无。结果提示ＮＥ促进ＭｏｎＡＰＦ的机制可能涉及到Ｃａ＾２＋和ＰＫＣ，而与ＰＫＡ无关。     

树突状细胞研究中激光扫描共聚焦显微镜的应用进展

免疫应答是机体重要的防御功能 ,抗原呈递细胞 (Antigenpresentingcells ,APC)是启动特异性免疫应答的关键。树突状细胞 (Dendriticcells,DC)是体内最强的一类APC ,参与免疫应答的双重调控作用。激光扫描共聚焦显微镜 (Laserscanningconfocalmicroscope ,LSCM)由于具有高分辨率、高灵敏度、“光学切片”、三维重建、动态分析等特点 ,为基础医学与临床医学的研究提供了有效手段 ,使DC的研究更加细致和深入。本文就LSCM在DC的形态学、免疫荧光定位、功能学方面的应用最新进展作一综述。     

人的活化T细胞出现的T_4~+T_6~+双标记现象

OKT系列单克隆抗体的产生,促进了人T细胞亚类的划分。最初的报道认为,人的外周血T细胞可分为OKT_4~+(T_4~+)和OKT_8~+(T_8~+)细胞两个亚类,分别占T细胞总数的55～65%和30～40%。最近有报道指出,人的末梢血中存在少量的T_4~+T_8~+双标记细胞,在体外活化的T细胞,可以暂时     

Interactive Effects of Alpha Feedback and Instructional Set on Subjective State

The effects of EEG alpha feedback and instructional set were studied in 40 college students. Reported subjective experiences during feedback were examined experimentally in a factorial “drug-drug set” design. Alpha and no-alpha feedback were each paired with alpha and neutral instructions, in order to observe the individual and combined effects of alpha activity and instructional set. The results showed that for an “alpha experience” to occur, both alpha activity and alpha set are necessary; neither alone is sufficient. Theoretical considerations based on the Schachter and Singer (1962) drug model and some implications for alpha feedback research are discussed.     

THE EEG CHANGES ASSOCIATED WITH SMOKING

Computer analysis of EEG data, recorded under both resting and work conditions following smoking, was compared to the appropriate control data in 6 male subjects. Following digitizing, a Power Spectral Analysis was performed which revealed significant reductions in the peak alpha frequency component up to 20 min following smoking, during a visual task. Eyes open resting data showed a similar but not significant loss, after 8 min. No indications of increased fast activity were found. These results were related to comparable work on animals and humans. A suggestion was made as to the relevance of these changes.     

Beschreibung eines vereinfachten Makrophagenmigrationshemm-Testes zur Messung zellvermittelter Immunreaktionen beim Menschen

Zusammenfassung Ein direkter Makrophagenmigrationshemm-Test mit heterologen Zellen (DMMTH) wird in einer Agarose-Tropfentechnik ausgeführt. Eine Suspension aus Peritonealmakrophagen des Meerschweinchens und Humanlymphozyten wird in 0,2%iger Agarose aufgeschwemmt, die mit einer automatischen Dosiervorrichtung tropfenförmig auf Platten für den Leukozytenwanderungstest aufgetragen wird. Die aus den erstarrten Tropfen erfolgende Auswanderung der Makrophagen kann planimetrisch gemessen werden. Die Streubreite der Zellkulturen beträgt im Mittel 8,6%. Eine signifikante Wanderungshemmung durch die Freisetzung des Migrations-Inhibitions-Faktors wird beobachtet, wenn sensibilisierte Humanlymphozyten mit dem in die Agaroselösung eingebrachten spezifischen Antigen reagieren. An Mantoux-positiven Probanden wurde die Sensitivität des Testes gezeigt.     

EEG Correlates of the Afterimage of Visual Stimulation

Occipital EEG alpha activity following visual stimuli of varying intensity was studied with particular reference to the visual afterimage. Subjects each received three stimuli of differing intensity under two conditions: with and without reporting of afterimage onset and offset required. A third condition controlled for possible reporting effects on the EEG. Findings indicate that visual afterimages are a significant determinant of the duration of EEG alpha desynchronization following visual stimulation and that where visual stimuli are employed the duration of EEG alpha blocking is unsuitable as an indicator of the OR to such stimulation.     

TAP polymorphism does not influence transport of peptide variants in mice and humans

The major histocompatibility complex (MHC)-encoded transporter associated with antigen processing (TAP) delivers cytosolic peptides to the lumen of the endoplasmic reticulum (ER) for presentation by MHC class I molecules. For the rat, it has been demonstrated that TAP polymorphism results in the selection of different sets of peptides, the nature of the C terminus being of particular importance. Here, we investigated whether TAP polymorphism in mice and humans has functional consequences for transport of peptide sets variable at the C-terminal residues. Using cell lines of H-2^d, H-2^k, and H-2^dxk haplotype and a panel of human lymphoblastoid cell lines expressing eight different TAP alleles, we detected species-specific transport patterns, but no significant influence of TAP polymorphism on peptide selection. In addition, peptides with different core sequences were translocated to the same extent by different TAP. These results suggest that a major contribution of human TAP polymorphism to disease progression and autoimmunity is not very likely.     

Ag and IL‐2 immune complexes efficiently expand Ag‐specific Treg cells that migrate in response to chemokines and reduce localized immune responses

An intravenous administration of a high‐dose antigen (Ag) can induce immune tolerance and suppress the immune response, but the mechanism remains unclear. We recently proved that a combined i.v. administration of OVA and IL‐2‐anti‐IL‐2 Ab immune complexes (IL‐2 ICs) efficiently expands OVA‐specific Treg cells in the thymus and induces their migration into peripheral blood, by using OVA‐specific TCR Tg‐expressing DO11.10 mice. Here, we demonstrate that the expanded OVA‐specific Treg cells rapidly move into the air pouch after OVA injection in DO11.10 mice. The migration was inhibited by blocking the axis of a chemokine receptor, CCR2. Moreover, prior treatment with OVA and IL‐2 ICs enhanced OVA‐specific Treg‐cell migration and inhibited OVA‐induced delayed‐type hypersensitivity (DTH) reactions in the skin of BM chimeric mice with 15% of T cells expressing OVA‐specific TCR. Blocking the CCR2 axis reversed this suppression of DTH in these mice. Furthermore, prior treatment with OVA and IL‐2 ICs effectively reduced DTH reactions even in WT mice possessing only a very small population of OVA‐specific T cells. Thus, the treatment with Ag and IL‐2 ICs can efficiently expand Ag‐specific Treg cells with the capacity to migrate and reduce localized immune responses.     

人工髋关节无菌性松动失效的生物力学分析与诊断推理

目的研究人工髋关节置换术后无菌性松动失效的力学机理以及引发松动的具体原因,提出对临床中发生无菌性松动事件进行失效诊断的具体方法。方法从骨水泥层强度、界面微动、应力遮挡、磨损与骨溶解等生物力学角度对无菌性松动的成因进行研究,分析无菌性松动失效与产品、临床和患者等因素的关系,并研究翻修术前检测松动的方法。结果提出无菌性松动失效原因推理路线图,成功利用荧光透视分析(fluoroscopic analysis,FSA)技术在翻修术前对无菌性松动进行了在体测定。结论无菌性松动失效分析推理路线图可以帮助展开失效事件的原因挖掘,应用FSA方法可以对松动进行在体测定与确认,辅助医生开展人工髋关节置换术后无菌性松动失效的诊治。     

结核分枝杆菌抗原分析及免疫交叉反应研究

目的：筛选和鉴定结核分枝杆菌特异性和保护性抗原，研究结核杆菌的免疫反应特点，以探索结核病诊断和治疗的新途径。方法：采用超声破碎和滤膜抽滤的方法分别得到菌体蛋白和滤液蛋白，通过Western blot试验用结核杆菌的单克隆抗体及结核病人血清来检测蛋白样品，把发生阳性反应的蛋白在BECKMAN LF3200／多肽氨基酸序列测定仪上进行N末端序列分析，并用结核杆菌的单抗对自身抗原组蛋白进行了检测。结果：结核杆菌的31kD和30kD蛋白与结核杆菌的单抗及病人血清反应均呈阳性，但与正常小鼠血清和健康人血清反应呈阴性。31kD和30kD蛋白的N末端序列分别为：Ala Glu Val Asp Trp Leu Val Phe Ala Val和Phe Ser Arg Pro Gly Leu Pro Val Glu Tyr。结核分枝杆菌的单抗与自身抗原组蛋白能发生免疫交叉反应。结论：结核杆菌的31kD和30kD蛋白是免疫保护性抗原，对免疫交叉反应分子基础的进一步研究必将增加对结核免疫机理的了解。     

Measurement of antigen specific lymphocyte proliferation using 5-bromo-deoxyuridine incorporation An easy and low cost alternative to radioactive thymidine incorporation

The classical in vitro assay for the determination of cell mediated immune responses is the lymphocyte transformation test (LTT) in which cell proliferation is measured by incorporation of radioactive labeled thymidine (³H-TdR). The LTT assay using ³H-TdR is less suited for modestly equipped laboratories as it is costly, laborious and involves the need to handle radioactive isotopes and specialized equipment.

Here we describe an improved alternative LTT method which is capable of detecting specific cellular immune reactions (CMI) against (mycobacterial) antigens in vitro. This assay, the bromodeoxyuridine-ELISA LTT test, is simple, less expensive, reproducible and is as sensitive as the ³H-TdR test. The specific advantages of the test are a simple denaturation step and the fact that no radioactive isotopes are needed. The test is specifically suited for research laboratories in tropical countries which study CMI in those human infectious diseases where this arm of the immune response plays a pivotal role in the generation of immunity, e.g., in tubercolosis, leprosy and leishmaniasis.     

Cellular,serological, and molecular polymorphism of the class I and class II loci of the canine Major Histocompatibility Complex

This study was undertaken to determine the relationships between canine cellular and serological determinants and more recently described genes. Such relationships might reveal information about immunological reactivity or function of various proteins. To do this we studied the haplotypic associations of dog leukocyte antigen (DLA) class I and class II alleles determined from a panel of 14 DLA-D homozygous dogs. This panel of dogs was typed for the serological determinants DLA-A, DLA-B and DLA-C. Polymorphisms for DLA-DQA1, DLA-DQB1, DLA-DRB1 and DLA-88 were also determined. The number of alleles (one or two) for two microsatellite markers in the DLA region were also determined. Analyses of the nucleotide sequences and of the serological and cellular typing data revealed that phenotypic homozygosity, as defined by the DLA-D type in mixed leukocyte culture (MLC), tended to correlate with homozygosity at the DLA-DRB1 locus but not necessarily at the DLA-DQB1 locus. Furthermore, MLC specificity was determined by other loci besides DLA-DRB1 and DLA-DQB1. The amino acid at position 63 of the DR beta chain could contribute to the DLA-B serological specificity. DLA-88, the most polymorphic class I gene characterized to date, did not have an easily identifiable association with either the DLA-A or DLA-C class I serological specificities. Homozygosity or heterozygosity of each of two microsatellite markers, FH 2200 and FH 2202, located in the class I or class II region, respectively, did not correlate with homozygosity or heterozygosity of the most polymorphic known class I (DLA-88) or class II (DLA-DRB1) genes.     

HLA-G and susceptibility to develop celiac disease

The Human Leukocyte Antigen-G has immunomodulatory function and its expression has been associated with several diseases. In our study we analyzed HLA-G polymorphisms in order to evaluate their possible association with susceptibility to celiac disease development. A total of 420 celiac patients and 509 controls were genotyped for HLA-G polymorphisms. We sequenced 800 bp upstream the ATG codon (5′ upstream regulatory region) and the whole 3′ untranslated region of the HLA-G gene, whereas the ΔC deletion at exon 3 was detected by RFLP-PCR.Five polymorphisms (namely −477 C>G, −369 C>A, 14 bp del/ins, 3187 A>G, 3196 C>G) and one haplotype (TCGGTACGAAITCCCGAG) were significantly more frequent in celiac patients than controls and associated with increased disease susceptibility. The 14 bp I/I, 3187 G/G, 3196 G/G genotypes and TCGGTACGAAITCCCGAG haplotype, were still significantly associated with increased disease susceptibility (and in addition also the 3003 C/C genotype) when the analysis was restricted to patients and controls presenting the DQ2.5 or DQ8 HLA-DQ celiac disease risk haplotypes.Our findings indicate an association between HLA-G gene polymorphisms and susceptibility to celiac disease development, suggesting that HLA-G molecule is possibly involved in the pathogenesis of the disease.     

检测抗HIV-1/2型抗体的双抗原夹心方法的建立及其试剂盒的研制

目的 建立更敏感的检测人免疫缺陷病毒（HIV）抗体的方法,并研制检测试剂盒。方法 根据HIV－1／2型的基因序列及其所编码氨基酸结构,采用固相法合成了HIV－1型的gp41.1、gp41.2、gp120、p24和HIV－2型的gp36五条多肽,混合包被酶标板作为固相抗原。用辣根过氧化物酶标记以上多肽抗原作为标记物,建立检测血清中抗HIV－1／2抗体的双抗原夹心ELISA法。同时,应用该方法制备检测HIV抗体的试剂盒,并检测三批中国卫生中药品和生物制品检定所HIV诊断试剂国家参比品。结果 建立了检测HIV－1／2抗体的双抗原夹心法。用检定所参比品检测,该方法特异性、灵敏度均为100％,变异系数小于10％。与间接法相比较其灵敏度、特异性均高于间接法（P＜0．05）。检测210份其他病种患者血清均为阴性。与GBI公司的HIV抗体诊断试剂比较,检测40份卫生部药品和生物制品检定所提供的质控参比品（阳性20份,阴性20份）,GBI试剂阴、阳性符合率及总符合率分别为100%(20/20)、85％（17／20）及92．5％（37／40）,而应用该方法所研制的诊断试剂盒、阳性符合率及总符合率为100％。该试剂已通过国家卫生部质检。与雅培公司HIV诊断试剂比较检测90份献血员血清和88份HIV－1／2型感染者血清,符合率为100％。试剂盒于37℃放置4d后的检测结果的阴、阳性判定不受影响。结论 本法特异性强、灵敏度高、稳定性好,适用于献血员的筛选和临床HIV感染的检测。     

PON2单克隆抗体的制备与初步鉴定

目的:制备PON2(paraxonase2)单克隆抗体(mAb),并进行初步鉴定.方法:利用生物信息学方法分析人类PON2蛋白序列,选取与小鼠同源性低,而免疫原性与亲水性均较强的片段,构建重组表达质粒pGEX-4T-1-PON2和PET-32a-PON2,GST-PON2和HIS-PON2融合蛋白在大肠杆菌中进行表达, 以HIS-PON2作为免疫原制备鼠mAb,以GST-PON2作为筛选抗原.采用Western blot、间接免疫荧光鉴定mAb的特异性.结果:GST-PON2和HIS-PON2融合蛋白均在大肠杆菌中获得高效表达,经常规的细胞融合和筛选获得2株可稳定分泌抗PON2的杂交瘤细胞株,这2株抗体可以识别HepG2细胞中的靶蛋白.结论:成功制备出2株抗PON2的mAb,并通过免疫荧光技术检测了该蛋白在HepG2细胞中的分布,为进一步进行PON2蛋白的的研究提供了有效的工具.     

血栓素B_2〔~(125)I〕放射免疫分析及其临床应用

本文报告~(125)I-血栓素B_2(TXB_2)放射免疫分析及其临床应用。TXB_2抗血清与其它前列腺素的交叉反应均小于0.42％,亲和常数为2.44×10~(10)M~(-1)。冷回收率94.7±7～106.0±13.1％,批内差异6.4％,批间差异11.8％,标准曲线测定范围2.5～160.0Pg/管。人血浆正常值143.5±8.8Pg/mL,尿正常值:229.9±14.4Pg/min。同一份样品用~(125)I和~3H两种放射免疫法测定,二者相关性良好。应用本法对冠心病、心肌梗塞患者及服阿司匹林前后的血浆和尿样品中TXB_2含量进行了探讨。