手工微量RhE抗原检测法应用体会

我们自行设计了手工微量RhE抗原检测法[1],该方法中增加了低离子介质溶液。该溶液可以增强抗原抗体的敏感性,促使抗原抗体复合物的形成[2]。尤其对弱抗原的检测结果更易判断,且检测时间短,敏感性高。现将我的应用体会总     

免疫组化中抗原修复方法及修复液的比较

目的本实验通过比较几种常用的抗原修复技术在免疫组化中的效果,寻求一种简洁、方便、实用的抗原修复方法。方法分别选取10种细胞核及细胞浆或膜标记的抗体,对理论上阳性的组织进行标记,对每种抗体均分别采用不修复,PH6.0柠檬酸煮沸,PH6.0柠檬酸高温高压修复,PH9.0EDTA煮沸,PH9.0EDTA高温高压修复。结果本实验的20种抗体均标记理论上为阳性的组织,不修复的有16种抗体阴性,水煮热修复与高压热修复的修复效果并无明显差别。抗原修复液中高PH值（9.0）时的染色结果明显好于相同修复方式的低PH值（6.0）。结论免疫组化质量与是否进行抗原有关,但与抗原修复方法无关;抗原修复液PH值影响免疫组化质量,掌握抗体最佳的修复液能提高免疫组化结果的阳性率、染色强度,稳定性及准确性。     

人类免疫缺陷病毒检测及临床意义

人类免疫缺陷病毒(HIV)检测必须严格按国家制定的《艾滋病检测工作管理办法》和《艾滋病检测技术规范》进行,整个实验过程中应有严格的质量保证体系。做好实验前、中、后的质量控制工作。通过HIV特异性抗体、抗原、核酸及病毒分离检测来确定HIV感染,WHO和我国卫生部推荐使用HIV感染诊断方法,是血清学抗体检测。1HIV抗体初筛检测1.1阿克苏夹心法原理将HIV抗原包被固相载体上,标本中HIV抗体与包被的HIV抗原结合形成复合物,利用抗体的多价性。又与酶(HRP)标记的同类HIV抗原即酶标板内的球状复合物,形成一个HIV抗原抗体酶标记的H…     

产前检查抗-M抗体检测方法的改进

本案例在做实验的过程中发现在检测免疫性抗-M抗体时,适当的调整检测方法能提高抗体检测率,对提早发现并预防相关MNss血型系统的溶血病有一定的意义。本实验首先通过直接凝集和抗人球蛋白的方法进行抗体筛选及抗体鉴定试验,为进一步确定不规则抗体的特异性,选择相应抗原阳性和阴性的进口谱细胞各两组进行确认试验,同时对患者红细胞的相应抗原进行检测,确认患者为NN型红细胞,含有不规则抗-M抗体,并对抗-M抗体的效价进行了检测。在检测的过程中发现对于免疫性的IgG类的不规则抗-M抗体,在通过DiaMed卡检测时,调整孵育温度,通过试管中抗人球蛋白法时调整洗涤的介质及孵育时间,均能有效的提高抗-M抗体的检测强度。     

噪声对实验动物免疫活性的影响

选37支雄性大白鼠进行实验研究。实验组遭受噪声强度为65分贝的作用,按一周5天,每天5小时共进行2个月实验。结束后将9支大白鼠处死,取血清测定自然免疫指标,包括杀菌力、溶菌酶、β—溶解素、正常抗体的补体及抗组织的补体结合的自体抗体。用酒精伤寒菌苗及 vi—抗原,使9支动物作一次腹膜内的免疫。接种后经7、14及21天,取尾静脉血测定伤寒菌的 O 和 Vi—抗原的抗体滴定度。用未遭受     

在广州市实验大白鼠中发现流行性出血热病毒抗原和抗体

1984年8~10月,作者对广州地区实验大、小白鼠EHF病毒感染情况进行了调查研究,共采集大白鼠296只,每只都采集了肺组织标本,EHF病毒抗原阳性率为6.08%。296只大白鼠中有233只在采集肺组织标本的同时采集了血液标本,EHF病毒抗体阳性率为13.73%。同时采得肺组织和血液标本的233只大白鼠中,EHF病毒抗原或/和抗体阳性者32只,感染率为11.49%,其中抗原抗体同时阳性者16只,阳性率为6.87%,抗原阴性抗体阳性者16只,阳性率为6.87%,未发现抗原阳性而抗体阴性者。发现雌性大白鼠EHF病毒感染率显著的高于雄性大白鼠。对大白鼠感染的EHF病毒来源进行了讨论。采集小白鼠786只,肺组织EHF病毒抗原全部阴性,未采集血液标本检测EHF病毒抗体。     

脂质体在免疫学中的应用、现状和展望

脂质体（Liposome）是一种人工合成的脂质膜性小球，1965年由Bansham等首先描述，其结构类同生物膜。近年来，它作为载体微球，在医学研究中应用广泛，不仅可作为活质分子，加药物、酶、离子等的载体，使其有选择地作用于靶器官．集中发挥更有效的作用。此外，它还可用于提高体外抗原一抗体反应的敏感性和可检测性。因此，它在免疫学中的应用已受到学者们越来越多的重视。l脂质体在免疫学中的应用脂质体作为抗原／抗体的载体微球，无论在体内的免疫学活性方面还是在体外的抗原一抗体反应中，均有重要的作用。11体内应用脂质体在体内的免疫…     

应用单克隆抗体对血吸虫循环抗原的免疫学特性化和检测

检测循环抗原比检测循环抗体更有可能改进血吸虫病的免疫诊断,很多研究已述及循环抗原的特性化和检测。阳极循环抗原和阴极循环抗原是其中两种主要的循环抗原,已证实为了特性化和检测这两种抗原去制备特异的多价抗血清是困难的,因此,我们采用了感染的和免疫的小鼠脾细胞制备单克隆抗体的模式,对血吸虫多糖抗原的抗体应答进行了测定,发现一些单克隆抗体主要是针对肠相关     

热处理的Ⅰ型嗜肺军团菌细胞抗原作酶联免疫吸附试验检测抗体的应用

菌细胞为抗原的酶联免疫吸附试验(BCA-ELISA),不用提取抗原,可测定直接作用于宿主体内的菌细胞表面裸露抗原决定簇(蛋白质、脂多糖)激起的抗体,能更确切地反映机体的免疫状态,其敏感性较肥达氏等细菌凝集试验测得的这种抗体滴度高10～100倍,已用于检测许多细菌表面(或菌毛)抗原的抗体[1-4].     

SEC_1酶免疫电极的研究

以酶、抗体相结合作为生物传感器的识别元件进行抗原的检测，不仅在临床医学上，而且在生化战剂的侦检中都有着广阔的应用前景。本文所研究的是将镀铂玻碳电极以化学吸附和交联的方式固定化葡萄糖氧化酶（ＧＯＤ），然后进一步耦联葡萄球菌肠毒素Ｃ１型抗体（Ａｎｔｉ－ＳＥＣ１），从而构成一种新型的电流型ＳＥＣ１酶免疫传感器。当抗原（ＳＥＣ１）与抗体发生免疫反应而形成抗原－抗体络合物后，使与抗体耦联的酶的活性受到调制，电极的响应信号发生变化，从而达到检测抗原的目的。实验表明，以化学吸附和交联两种方式制得的酶免疫电极在检测抗原时，与响应电流存在很好线性关系的浓度范围分别为２０～１００μｇ／Ｌ、３～１８μｇ／Ｌ。     

Monopoly Interest Called Root of Staff Privilege Denials

In a number of infections (for example, tuberculosis) acquired resistance depends on a cell-mediated type of immunity rather than the classic antigen-antibody reaction. The mediating cell is a short-lived small lymphocyte, produced specifically in response to antigenic stimulus. In turn it mobilizes phagocytes at the infective site and activates them to become much more effective antibacterial agents.     

Prospects and advancements in C-reactive protein detection

C-reactive protein (CRP) is one of the earliest proteins that appear in the blood circulation in most systemic inflammatory conditions and this is the reason for its significance, even after identification of many organ specific inflammatory markers which appear relatively late during the course of disease. Earlier methods of CRP detection were based on the classical methods of antigen-antibody interaction through precipitation and agglutination reactions. Later on, CRP based enzymatic assays came into the picture which were further modified by integration of an antigen-antibody detection system with surface plasma spectroscopy. Then came the time for the development of electrochemical biosensors where nanomaterials were used to make a highly sensitive and portable detection system based on silicon nanowire, metal-oxide-semiconductor field-effect transistor/bipolar junction transistor, ZnS nanoparticle, aptamer, field emission transmitter, vertical flow immunoassay etc. This editorial attempts to summarize developments in the field of CRP detection, with a special emphasis on biosensor technology. This would help in translating the latest development in CRP detection in the clinical diagnosis of inflammatory conditions at an early onset of the diseases.     

Serum Complement and Chronic Glomerulonephritis

Biopsy studies are differentiating several types of glomerular injury in patients with primary persistent glomerulonephritis, and in a majority serial measurements show serum complement levels to be normal rather than low (as tradition holds). These findings suggest that mechanisms other than, or in addition to, the antigen-antibody reaction may be responsible for the condition classically known as “chronic nephritis.”     

Apoptosis of human tumor cells by chemotherapeutic anthracyclines is enhanced by Bax overexpression

One of the major factors for efficacy of a chemotherapeutic drug is its activity to induce apoptosis of tumor cells. Doxorubicin and daunorubicin, radiomimetic anthracycline-group drugs, have been used for chemotherapy for about 30 years. Here we established the colorectal tumor and osteosarcoma cells in which Bax expression can be induced by the treatment of isopropyl-beta-D-thiogalactopyranoside, and examined the effect of the Bax overexpression on the cell death caused by these drugs. While the Bax overexpression neither affected growth nor morphology of the undamaged cells, it enhanced the cell death caused by these drugs. Increase in cellular nucleus fragmentation and DNA ladder formation indicates that the Bax-enhanced cell death is due to enhanced apoptosis of the drug-treated cells. The enhanced cell death was not observed when the cells were irradiated with X-ray or treated with other chemotherapeutic agents we examined. These results indicate that Bax may have a specific role to enhance the efficacy of chemotherapy with anthracycline-group agents.     

Contrasting Effects of Heavy Metals on Sponge Cell Behavior

Mediterranean coastal areas are highly contaminated by heavy metals, which have been reported to produce harmful effects in marine organisms. Sponges are particularly vulnerable to waterborne metals because they are able to process large amounts of water. Dissociated sponge cells can move in response to external stimuli, and the cell body changes shape through production of pseudopodia and phylopodia. We studied for first time the effects of heavy metals (cadmium copper and mercury) on motility and aggregation of isolated sponge cells. Cell shape was assessed by using several shape indices. The three metals studied induced changes of different sign on cell shape. Mercury arrested movement of sponge cells, which tended to be rounded, without pseudopodia. In contrast, moderate concentrations of copper and cadmium enhanced pseudopodia formation and cell motility. On the other hand, the three metals enhanced cell aggregation at the concentrations assayed. Our results show that sponge cells respond to metal pollution in different ways and that these responses can be assessed by calculating several shape indices.     

Detection of antimicrofilarial ES antigen-antibody in immune complexes in Bancroftian filariasis by enzyme immunoassay

An enzyme immunoassay using penicillinase conjugated to Wuchereria bancrofti microfilarial ES antigen has been developed to detect specific antibody in circulating immune complexes in Bancroftian filariasis. Immune complexes were prepared by 3% polyethylene glycol (PEG) precipitation. 44 sera belonging to different groups were tested. 16 of 19 clinical filarial and two of 16 endemic normal sera but none of the non-endemic normal sera showed the presence of antimicrofilarial ES antigen-antibody in immune complexes.     

CD40 antibody as an adjuvant induces enhanced T cell responses

Monoclonal antibodies against CD40, conjugated to antigen, act as potent immunological adjuvants for primary antibody responses. We show here that CD40mAbs can also act as strong adjuvants for memory antibody responses, and for T cell responses as measured by ex vivo T cell proliferation to antigen, and delayed type hypersensitivity. Interferon gamma secretion in response to antigen is also enhanced. Finally, the adjuvant effect of CD40mAbs for secondary antibody responses is transferred with T cells rather than B cells. CD40mAb apparently have potent adjuvant effects on both Th1-like cells, and on T cells able to promote B cell antibody production. It is possible that the adjuvant effects of CD40 are mediated at least in part, indirectly, through enhanced antigen presentation by specific B cells, to T cells.     

Retinoids and Carotenoids as Inhibitors of Carcinogenesis and Inducers of Cell-Cell Communication

In cell cultures of carcinogen-initiated fibroblasts, a number of retinoids and carotenoids can reversibly inhibit progression to the transformed state. The anticarcinogenic action of retinoids and carotenoids is closely correlated with enhanced gap-junction cell-to-cell communication and with increased synthesis of the gap-junction protein, connexin.     

Adverse Benzo[a]pyrene Effects on Neurodifferentiation Are Altered by Other Neurotoxicant Coexposures: Interactions with Dexamethasone,Chlorpyrifos, or Nicotine in PC12 Cells

Background: Polycyclic aromatic hydrocarbons are suspected developmental neurotoxicants, but human exposures typically occur in combination with other neurotoxic contaminants.Objective and Methods: We explored the effects of benzo[a]pyrene (BaP) on neurodifferentiation in PC12 cells, in combination with a glucocorticoid (dexamethasone, used in preterm labor), an organophosphate pesticide (chlorpyrifos), or nicotine.Results: In cells treated with BaP alone, the transition from cell division to neurodifferentiation was suppressed, resulting in increased cell numbers at the expense of cell growth, neurite formation, and development of dopaminergic and cholinergic phenotypes. Dexamethasone enhanced the effect of BaP on cell numbers and altered the impact on neurotransmitter phenotypes. Whereas BaP alone shifted differentiation away from the cholinergic phenotype and toward the dopaminergic phenotype, the addition of dexamethasone along with BaP did the opposite. Chlorpyrifos coexposure augmented BaP inhibition of cell growth and enhanced the BaP-induced shift in phenotype toward a higher proportion of dopaminergic cells. Nicotine had no effect on BaP-induced changes in cell number or growth, but it synergistically enhanced the BaP suppression of differentiation into both dopaminergic and cholinergic phenotypes equally.Conclusion: Our results indicate that, although BaP can act directly as a developmental neurotoxicant, its impact is greatly modified by coexposure to other commonly encountered neurotoxicants from prenatal drug therapy, pesticides, or tobacco. Accordingly, neurodevelopmental effects attributable to polycyclic aromatic hydrocarbons may be quite different depending on which other agents are present and on their concentrations relative to each other.     

Early Treg suppression by a listeriolysin-O-expressing E. coli vaccine in heterologous prime–boost vaccination against cancer

Studies have shown that an enhanced CD8+ T cell response and better tumor protection can be achieved by heterologous prime–boost vaccination in mice. Such heterologous vaccination can be more immunogenic than the homologous setting. We previously demonstrated that a listeriolysin-O (LLO)-expressing E. coli vaccine can enhance CD8-cytotoxic T cell (CTL) responses by reducing regulatory T cell (Treg)-directed suppression. In the present study, we assessed the combination of this approach with plasmid DNA vaccination, in a prime–boost immunization strategy. E. coli-LLO bacteria expressing ovalbumin (OVA) and plasmid pcDNA-encoding OVA were used to vaccinate naive or B16-OVA tumor-bearing C57B6 mice. The anticancer activity was measured in a tumor prevention or therapeutic model. Higher OVA-specific CD8+ T cell responses and greater tumor inhibition were seen in the bacterial-prime/plasmid–boost setting than with the homologous and reversed sequences. This tumor protection effect from heterologous prime–boost remained in the therapeutic model. When examining the Treg effect during the prime–boost immunization, we found that only early Treg-suppression/depletion could lead to better antigen-specific CTL and tumor response. Our studies offer the first evidence that a listeriolysin-O E. coli vaccine can induce an enhanced antitumor effect in conjunction with DNA in a heterologous prime–boost protocol, and suggest that early Treg inhibition is crucial to a successful immunization against cancer.