从皮肤试验分析变应性鼻炎的诊断

目的分析变应性鼻炎的诊断。方法采用皮肤点刺试验检测1248例成人变应性鼻炎患者的变应原。结果517例(41.43%)患者呈阳性皮肤反应,位于前4位的变应原是:屋尘螨、粉尘螨、杂草及树。对1种或2种变应原过敏者占76.4%。对2种变应原过敏的患者,多为具有交叉抗原的变应原。变应原等级为( )以上者占75%。结论皮肤点刺试验是变应性鼻炎诊断的重要手段,可以作出明确的特异性变应原的种类及严重程度的诊断。     

碘硒对大鼠腹腔巨噬细胞抗原提呈作用的影响

目的 通过观察碘硒对大鼠腹腔巨噬细胞抗原提呈作用的影响,初步探讨两者对自身免疫性甲状腺疾病发病的影响及其免疫学机制.方法 选用雌性Lewis大鼠20只,根据随机抽样的原则,分为4组:低硒适碘组(LSeN1),低硒高碘组(LSeH1),适硒适碘组(NSeN1),适硒高碘组(NSeH1).各组用人工合成的低硒低碘饲料和饮用含不同浓度硒或碘的去离子水(用K103和Na,SeO3配置)喂养3个月.制备各组大鼠腹腔巨噬细胞及卵白蛋白(OVA)致敏的T细胞,将两者共同培养,进行抗原提呈实验,采用ELISA方法测定上清中IL-2水平;采用RT-PCR方法检测各组脾细胞共刺激分子CD86 mRNA的表达水平.结果 NSeH1组培养上清中IL-2水平为(43.22±3.27)pg/ml明显高于NSeN1组IL-2水平(25.74±2.45)pg/ml.LSeN1组IL-2水平为(15.79±2.13)pg/ml明显低于NSeN1组.NSeH1组大鼠脾细胞CD86mRNA表达水平(CD86/βp-actin:0.52±0.10)明显高于NSeN1组(CDB6/13.actin:0.35±0.04).结论 高碘使巨噬细胞的抗原提呈作用呈现高于正常状态,成为诱发甲状腺自身免疫发生的一个重要的因素.低硒可使大鼠腹腔巨噬细胞对OVA抗原识别和提呈作用减弱,维持免疫自稳机制失调,可能也是构成诱发自身免疫病的一个因素.     

乳腺癌中CXCR4的表达及与PCNA、淋巴结转移的关系

目的:探讨趋化因子受体CXCR4在乳腺癌中的表达及与PCNA、淋巴结转移的关系。方法:S-P法检测乳腺癌、乳腺增生组织及乳腺纤维腺瘤中CXCR4表达,分析CXCR4在乳腺癌中的表达与患者淋巴结转移状态、PCNA相互关系。结果:CXCR4在乳腺癌组织中表达率为59.4%(38/64),与乳腺纤维腺瘤组织有显著性差异(P<0.05),与患者淋巴结转移状态、PCNA正相关(P<0.05)。结论:乳腺癌中CXCR4高表达,更易发生淋巴结转移及较高PCNA表达,有望成为新的乳腺癌治疗靶点。     

Allergenicity of Common Foods Restricted in Respiratory Allergy

Although hypersensitivity to foods is often linked to exacerbations of symptoms of respiratory allergy, no such information is available regarding the foods traditionally considered to play a probable etiological role in respiratory allergy in India, which are in fact quite different from the ones implicated in the West. The present study was undertaken to investigate whether the practice of withholding certain common foods by parents and practitioners of indigenous systems of medicine (i.e. Ayurvedic and Unani systems of medicine) in children suffering from respiratory allergy had any scientific basis or explanation as judged by modern techniques of investigation. Skin prick tests were performed on 64 children with symptoms pertaining to respiratory allergy (32 each in study and control group) using crude antigenic food extracts. Oral food challenges were administered to children to confirm or rule out allergenicity of food (s) incriminated on the basis of the clinical history and/or a positive skin test. Parental history of food restriction alone, in absence of positive skin prick test was of little value in predicting a positive response to the food challenges (1 challenge positive out of 77 based on food restriction: 1.29%). Only 27.02% and 18.75% of positive skin tests were found to be clinically significant in study and control groups respectively. Traditionally, food beliefs were upheld in only 12.5% children for immediate onset clinical reactions (with 5.31% of the foods restricted in their diet) and 9.37% children for delayed onset clinical reactions (with 3.19% of the foods restricted in their diet). The present study shows that even though food restriction is a common practice in patients with respiratory allergy in India, objective documentation of Type I reactions due to these foods cannot be obtained in a majority of such children.     

肺瘤平膏及其拆方对树突状细胞抗原递呈功能影响的分子机制研究

目的：观察肺瘤平膏及其拆方干预体外培养体系中DC抗原递呈功能的作用并探讨其分子机制。方法：建立体外人血PBMC诱导分化成熟DC的诱导培养体系,从健康人外周血中分离获得PBMC,采用多种细胞因子（TNFα、IL-4、GM-CSF）联合诱导,获得了分化与功能相对成熟的DC,并采用流式细胞仪检测技术,对肺瘤平膏及其拆方含药血清干预成熟DC与抗原递呈功能相关表面分子表达,及其对DC分泌IL-12水平影响进行初步研究。结果：肺瘤平膏可上调DC与抗原递呈功能相关膜分子MHC-Ⅱ、CD80、CD83、CD86及CD40的表达,并促进DC分泌IL-12水平,其方中扶正部分的作用不容忽视。结论：在扶正培本治则指导下的肺瘤平膏能够调节DC抗原递呈功能,提高机体的抗肿瘤免疫监视功能。     

中老年男性692例血清前列腺特异性抗原检测分析

目的:探讨中老年男性血清前列腺特异性抗原(PSA)浓度与年龄、生活习惯的关系.方法:以692例中老年男性为研究对象,取样后分离血清,采用双抗体夹心法,根据标准曲线计算出样品的浓度;生活习惯采取问卷调查的方式.结果:血清PSA浓度随年龄的增长逐渐升高,并且与生活习惯有一定关系. 结论:血清PSA浓度与年龄呈正相关,一些不良生活习惯可使血清PSA升高.     

Isolation of potent human Fab fragments against a novel highly immunogenic region on human muscle acetylcholine receptor which protect the receptor from myasthenic autoantibodies

In the autoimmune disease myasthenia gravis (MG), antibodies against the muscle nicotinic acetylcholine receptor (AChR) cause loss of functional AChR in the neuromuscular junction. To isolate AChR-specific human antibody fragments (Fab), a phage-display library was constructed from an MG patient's thymic B lymphocytes. The first Fab isolated had a low affinity for human AChR, but two sequential antibody chain shufflings using the MG donor heavy and light chain gene repertoires resulted in isolating two new Fab with an approximately 30-fold higher binding ability. The selected Fab contained extensively mutated heavy and light chains and probably represent intraclonal variants of a common progenitor having diverged in vivo by somatic hypermutation. Interestingly, the isolated Fab bound to an extracellular highly immunogenic region located either on an alpha-subunit site affected by the gamma/epsilon-subunits or on the interface between alpha- and gamma/epsilon-subunits. This region is not the previously described "main immunogenic region" (MIR), although it seems to be close to it, as one improved Fab and an anti-MIR mAb competed for AChR binding with distinctly different subpopulations of MG sera. Furthermore, this Fab protected surface AChR in cell cultures against MG autoantibody-induced antigenic modulation, suggesting a potential therapeutic use in MG, especially in combination with a human anti-MIR Fab.     

Development of a cluster of differentiation antibody-based protein microarray

Protein microarrays combine aspects of DNA microarrays and ELISA for the parallel interrogation of a biological sample using a multiplex of protein biomarkers. Here we report the development of a protein microarray consisting of a subset of CD antibodies and CRP. Several preparations (culture supernatant, ascites fluid and purified Ig) of each antibody were used in a forward phase protein microarray. Microarrays were fabricated using a non-contact printer delivering 300 pL (+/-30 pL) to specific locations on polyacrylamide gel-based substrates. Following production, microarrays were blocked for non-specific binding and incubated with sera conjugated directly with Cy3. Using CRP as a control biomarker, 12 clinical samples (inflammatory conditions and controls) were interrogated using the protein microarray format and results compared to CRP measured by conventional immunoassay. The data obtained from the microarray correlated with CRP assessed by immunoassay. Subsequently CRP 'positive' samples were interrogated for CD antigen expression; which revealed CD25 and CD45RO expression in all samples. Whilst this study focussed on a subset of CD antibodies, it is anticipated that this array could be expanded to include a larger number of CD antibodies and allow screening of sera from multiple conditions in order to identify disease markers.     

HLA type-independent generation of antigen-specific T cells for adoptive immunotherapy

Adoptive immunotherapy with antigen-specific T cells has been successfully used to treat certain infectious diseases and cancers. Although more patients may profit from T cell therapy, its more frequent use is restricted by limitations in current T cell generation strategies. The most commonly applied peptide-based approaches rely on the knowledge of relevant epitopes. Therefore, T cells cannot be generated for diseases with unknown epitopes or for patients with unfavorable HLA types. We developed a peptide-based approach for HLA type-independent generation of specific T cells against various proteins. It is based on short-time stimulation with peptide libraries that cover most CD4(+) and CD8(+) T cell epitopes of given proteins. The procedure requires no prior knowledge of epitopes because libraries are synthesized solely on the basis of the protein's amino acid sequence. Stimulation is followed by immunomagnetic selection of activated IFN-gamma-secreting cells and nonspecific expansion. To evaluate the protocol, we generated autologous T cells specific for a well-characterized antigen, the human cytomegalovirus phosphoprotein 65 (pp65). Generated T cell lines consisted of pp65-specific CD4(+) and CD8(+) lymphocytes that displayed antigen-specific killing and proliferation. The protocol combines the biosafety of peptide-based approaches with HLA type independence and may help to advance adoptive immunotherapy in the future.     

树突状细胞体外刺激对HBV特异性细胞毒T细胞影响的研究

目的探讨通过聚肌胞体外作用树突状细胞后改善慢性乙型肝炎患者树突状细胞功能,并在体外活化自身T细胞获得高频数HBV特异性细胞毒性T细胞(CTL)的方法。方法分离慢性乙型肝炎病人外周血单个核细胞,在粒巨噬细胞集落刺激因子(GM CSF)和白细胞介素4(IL4)的诱导下培养树突状细胞。培养的第7天加入聚肌胞刺激获得成熟树突状细胞,经HBVcore1827肽负载后与自身T淋巴细胞共培养,通过酶联斑点计数法(Elispot)及MHC肽四聚体法(Tetramer)比较病人T细胞未经自身树突状细胞刺激组、经HBVcore1827肽负载的自身树突状细胞刺激组、经聚肌胞促成熟的HBVcore1827肽负载的自身树突状细胞刺激组中HBV特异性的CTL的功能和频数。结果慢性乙型肝炎病人外周血单核细胞体外经GM CSF和IL4诱导可转化为树突状细胞,树突状细胞转化过程中聚肌胞的刺激可显著上调树突状细胞表面分子CD80、CD83的表达(P<0.01),促进树突状细胞的成熟。Elispot法检测分泌IFNγ的CTL的频数病人T细胞未经自身树突状细胞刺激组分泌频数为(9～28)/1×105T细胞,均值16;经HBVcore1827肽负载的自身树突状细胞刺激组频数为(30～67)/1×105T细胞,均值为46;经聚肌胞促成熟的HBVcore1827肽负载的自身树突状细胞刺激组频数为(59～130)/1×105T细胞,均值为98。三组数据