恶性淋巴瘤单克隆抗体的制备和初步鉴定

采用人恶性淋巴瘤细胞株Raji细胞作为免疫原免疫BALB/C小鼠, 按常规方法融合并经间接ELISA法筛选, 成功地制备了1株抗Raji细胞单克隆抗体(MAb), 命名为SZ138.用琼脂糖双扩散鉴定其为IgG1类, ELISA法测定其腹水效价为1:3.2×104.MAb流式细胞仪检测表明, 该抗体与Raji和Daudi呈阳性反应;与外周红细胞、白细胞(包括淋巴细胞和中性粒细胞)、未活化血小板、内皮细胞株ECV呈阴性反应.免疫组化SP法显示, MAb SZ138识别的抗原在胃肠道的腺体和肝脏有所表达, 而在心、脑、扁桃体、脾、脂肪组织、动脉血管壁、神经节、横纹肌、平滑肌等组织不表达或表达很低.Western blotting分析显示, MAb SZ138可特异识别还原条件下相对分子量为170kD的蛋白多肽.结果提示: MA SZ138对恶性淋巴瘤的诊断和治疗可能具有一定的临床意义.     

抗人白细胞单克隆抗体的特异性及其特性鉴定

目的制备抗人白细胞单克隆抗体（ｍＡｂ），并对其识别抗原及组织的特异性进行鉴定。方法采用分离的人全白细胞悬液免疫Ｂａｌｂ／ｃ小鼠，取脾细胞与Ｓｐ２／０细胞常规融合，用间接ＥＬＩＳＡ筛选ｍＡｂ，流式细胞仪及免疫组化染色ＳＰ法鉴定其组织特异性。　结果成功地制备１株抗人白细胞　ｍＡｂ，命名为ＳＺ－１０５。ＥＬＩＳＡ法测定其腹水效价为１×１０－５。琼脂糖双扩散鉴定为ＩｇＧ１类。流式细胞仪间接免疫荧光技术及免疫组化ＳＰ法测定表明，ｍＡｂ　ＳＺ－１０５可选择性地与外周血液的粒细胞、单核细胞和淋巴细胞呈强阳性反应，而与红细胞及血小板不出现反应。该ｍＡｂ还可与正常人骨髓的白细胞前体起反应。另外，在肝、肺、脾、胸腺及淋巴结正常组织中的巨噬细胞表面，也有ＳＺ－１０５抗原表达。ＳＺ－１０５抗原经亲和层析纯化，再经ＳＤＳ－ＰＡＧＥ　和Ｗｅｓｔｅｒｎ　ｂｌｏｔ证实，ｍＡｂ　ＳＺ－１０５识别的抗原是相对分子质量（Ｍｒ）为７５　０００左右的单链蛋白多肽。结论　获得１株具有高度免疫学活性和组织特异性的抗人白细胞ｍＡｂ　ＳＺ－１０５，其对研究白细胞的分化、免疫功能，以及隐匿性急、慢性炎症疾病的放免显像诊断都具有一定的临床意义。     

人成纤维细胞生长因子-21单克隆抗体的制备及抗原表位的确定

目的 制备小鼠抗人成纤维细胞生长因子(hFGF)-21单克隆抗体(mAb),通过细菌展示确定该mAb的抗原表位.方法 用hFGF-21作为检测和免疫抗原,间接ELISA筛选分泌抗人hFGF-21 mAb的杂交瘤细胞株；用FITC标记该mAb,并克隆hFGF-21的不同片段到展示载体Apex上,通过流式细胞仪筛选其抗原表位.结果 成功筛选出1株抗hFGF-21抗体的细胞株,其分泌抗体重链的亚型为IgG 2b,轻链为Kappa链；该杂交瘤细胞株腹水的效价为1:4.096×106；传30代及液氮中保存3个月,该细胞株能稳定分泌抗hFGF-21 mAb,且效价稳定；Western blot法检测证明该抗体与人FGF-21有很好的特异性；该mAb与小鼠FGF-21有交叉反应；通过流式细胞仪对抗原表位的筛选,该mAb可与hFGF-21下游的第107～121个氨基酸反应.结论 成功的制备出特异性、高稳定性的小鼠抗hFGF-21 mAb,确定了该mAb的抗原表位在第107～121位氨基酸.     

抗人分泌性白细胞蛋白酶抑制剂单克隆抗体的制备与鉴定

目的制备抗人分泌性白细胞蛋白酶抑制剂(hSL-PI)单克隆抗体(mAb)。方法以hSLPI为免疫BALB/c小鼠,运用杂交瘤技术制备抗hSLPI mAb。采用Ig类与亚类鉴定试剂盒鉴定mAb的Ig亚类;通过ELISA、Western blot、免疫组化染色法、流式细胞术、激光共聚焦显微镜技术鉴定mAb的特异性。结果获得4株可稳定分泌抗hSLPI mAb的杂交瘤细胞,其分泌的mAb均为IgM。Western blot分析表明,此株mAb所识别的靶分子的相对分子质量(Mr)约为12000。免疫组化染色表明,mAb与肺和结肠组织的上皮细胞、疑似肥大细胞、扁桃体和包皮组织的疑似肥大细胞的反应性均呈阳性。流式细胞术分析显示,4株mAb与人肺腺癌细胞系A549细胞中的SLPI均呈阳性反应。激光共聚焦扫描显微镜观察,荧光标记的阳性反应物主要位于A549细胞的胞质内。结论成功地制备抗hSLPI mAb,为变态反应性疾病和炎症性疾病的研究工作提供了有用的试剂。     

烯脂酰辅酶A水解酶单克隆抗体的制备与鉴定

目的:制备小鼠抗人烯脂酰辅酶A水解酶(enoyl CoA hydratase,ECHS1)的单克隆抗体(mAb)并进行初步鉴定.方法:将正常成人肝组织匀浆离心并分离线粒体,用线粒体总蛋白免疫BALB/c小鼠,采用杂交瘤技术制备mAb,并通过间接ELISA、Western blot、免疫沉淀、免疫组化和间接免疫荧光等方法对mAb进行特异性鉴定,通过Uni-ZAP XR正常成人肝脏cDNA表达文库筛选鉴定抗原.结果:获得1株可稳定分泌抗人ECHS1 mAb 的杂交瘤细胞株BGB095.该mAb的Ig亚类(型)为IgG1(κ),可用于ELISA检测、Western blot、免疫沉淀、免疫组化等实验.结论:成功制备了抗人ECHS1的mAb,为代谢疾病和肿瘤的研究提供了有力的工具.     

抗人LSECtin单克隆抗体的制备与初步鉴定

目的:制备抗肝脏、淋巴结窦内皮细胞C型凝集素(LSECtin)单克隆抗体(mAb),并进行特性鉴定。方法:采用原核表达的LSECtin免疫BALB/c小鼠,以间接ELISA法筛选分泌特异性mAb的杂交瘤细胞,采用蛋白印迹、间接免疫荧光、流式细胞术和免疫组化染色法鉴定mAb的特异性。结果:共获得8株可稳定分泌mAb的杂交瘤细胞株。mAb的Ig亚类均为IgG,效价达1∶106~1∶107。这些mAb均可识别转染3T3细胞膜上的人LSECtin,6株mAb可特异识别肝脏窦内皮细胞。结论:成功地制备8株抗LSECtin的mAb,经免疫印迹、流式细胞术和免疫组化染色检测,这些mAb的特异性良好,为研究LSECtin的功能提供了有力的试剂。     

抗二羰基/L-木酮糖还原酶单克隆抗体的制备与鉴定

目的制备抗人二羰基/L-木酮糖还原酶(Dicarbon-yl/L-xylulosereductase,DCXR)的单克隆抗体(mAb)并进行初步鉴定。方法将正常成人肝组织匀浆离心并分离线粒体,用线粒体总蛋白免疫BALB/c小鼠,采用杂交瘤技术制备mAb,并通过间接ELISA法、Westernblot及免疫组化的方法对mAb进行特性鉴定,通过免疫沉淀联合质谱、Uni-ZAPXR表达文库筛选鉴定抗原。结果获得1株可稳定分泌抗人DCXRmAb的杂交瘤细胞系。mAb的Ig亚类(型)为IgG1(κ),该mAb可用于ELISA检测、Westernblot、免疫组化和免疫沉淀实验。结论成功制备了抗人DCXR的mAb,为DCXR的研究提供了有力的工具。     

抗结肠肿瘤相关抗原单克隆抗体的制备及鉴定

目的:制备抗结肠肿瘤相关抗原的单克隆抗体(mAb)并对其进行初步的鉴定及功能研究。方法:将结肠癌细胞系或磨碎的结肠癌组织免疫小鼠,按常规方法进行细胞融合,采用活细胞周围花环形成的方法进行阳性克隆的筛选。用间接免疫荧光染色和流式细胞术分析及酶免疫组织化学的方法进行初步鉴定,并用抗体介导的补体依赖的细胞毒试验(CDC)进行初步的功能研究。结果:获得了2株分泌抗结肠肿瘤相关抗原mAb杂交瘤细胞株3A12和6A8。流式细胞仪检测,2株杂交瘤分泌的抗体均能在结肠癌细胞系中均为阳性。普通石蜡切片及组织芯片的免疫酶组织化学染色结果,显示2株mAb均在结肠肿瘤组织中有较高的阳性率。2株mAb分别与靶细胞及补体作用后,对于靶细胞的杀伤率分别为98.1%和42.4%。结论:成功地制备了2株识别结肠癌细胞表面抗原的mAb,在结肠癌的诊断和治疗中可能具有潜在的应用价值。     

抗丙酮酸脱氢酶复合体E2亚基单克隆抗体的制备与鉴定

目的 制备抗人丙酮酸脱氢酶复合体E2亚基(pyruvate dehydrogenase complex E2,PDC-E2)的单克隆抗体(mAb)并进行初步鉴定.方法 将正常成人肝组织匀浆离心并分离线粒体,用线粒体总蛋白免疫Balb/c小鼠,采用杂交瘤技术制备mAb,并通过间接ELISA法、Western blot及免疫组化的方法对mAb进行特性鉴定,通过免疫沉淀联合质谱、Uni-ZAP XR肝脏cDNA表达文库筛选鉴定抗原.结果 获得1株可稳定分泌抗人PDC-E2 mAb 的杂交瘤细胞系.mAb的Ig亚类(型)为IgG1(κ), 该mAb可用于ELISA检测、Western blot、免疫组化和免疫沉淀实验.结论 成功制备了抗人PDC-E2的mAb,为PDC-E2的研究提供了有力的工具.     

抗11β-羟类固醇脱氢酶1的单克隆抗体的制备与鉴定

目的:制备抗人11β-羟类固醇脱氢酶1(homo sapiens hydroxysteroid 11-beta dehydrogenase 1,HSD11B1)的单克隆抗体(mAb)并初步鉴定其特性.方法:将正常成人肝组织匀浆离心并分离线粒体,用线粒体总蛋白免疫BALB/c小鼠,采用杂交瘤技术制备mAb,并通过间接ELISA法、Western blot、免疫组化的方法对mAb进行特性鉴定,通过免疫沉淀、Uni-ZAP XR肝脏cDNA表达文库筛选鉴定抗原.结果:通过间接ELISA筛选获得1株可稳定分泌抗人HSD11B1 mAb的杂交瘤细胞系.其分泌的mAb的Ig亚类(型)为IgG1(κ),Western blot结果显示该mAb可以特异地识别相对分子质量(Mr)为35 000的蛋白;应用mAb CBF245对人肝脏cDNA表达文库(Uni-ZAP XR)进行筛选,阳性噬菌斑测序结果显示5个阳性克隆插入序列均为HSD11B1.结论:mAb BAD062特异地识别抗原HSD11B1,该mAb可用于ELISA检测、Western blot、免疫组化和免疫沉淀实验,为HSD11B1的研究提供了有力的工具.     

Renal dysplasia and asplenia in two sibs

A family is reported in which two sibs, one male and the other female, both died within 24 hours of birth with enlarged polycystic kidneys. Postmortem histology in the second child showed gross renal dysplasia. In both children the pancreas was enlarged, nodular and cystic but the liver appeared macroscopically normal. In the second child, histological examination confirmed pancreatic fibrosis with cystic dilation of ducts, but showed portal fibrosis with bile duct proliferation in the liver.
This combination of findings is very reminiscent of those in a girl and her brother reported by Ivemark et al. (1959). The children reported here also showed absence or hypoplasia of the spleen, cardiac anomalies and other features of the Ivemark syndrome (Ivemark 1955), a quite different, usually sporadic, congenital disorder. It is suggested that the children described here have a distinct lethal congenital disorder, probably inherited in an autosomal recessive manner.     

Schizophrenia in a North Swedish geographical isolate, 1900–1977. Epidemiology, genetics and biochemistry

Over 200 schizophrenic patients belonging to three major and interrelated pedigree complexes have been investigated over the past 30 years in a North Swedish geographically isolated population, presently numbering about 6,000. An intensive investigation of a number of biochemical correlates and genetic markers in a few selected families belonging to one of the major pedigrees has indicated new strategies for the current research program.
Schizophrenia, as defined operationally, is significantly associated with decreased activities of two enzymes (1) blood platelet monoamine oxidase, (2) plasma dopamine-β-hydroxylase, and (3) with the genetic marker Gc² (group specific antigen). Both enzymes are subject to genetic variation. A positive score for linkage between schizophrenia and low plasma DBH activity has been calculated, but, so far, available data are insufficient for discrimination between linkage and partial contribution of genetically controlled low plasma DBH to the pathogenesis of the disease. Alternatively, both mechanisms could be involved.
As a model for continued research, schizophrenia is explained as based on a double dominant-recessive genotype (Aabb), representing a vulnerability which in about 50 % of cases develops into clinical schizophrenia. It is suggested that the dominant mutation (A) operates on or affects MAO activity, and that the recessive genotype (bb) is instrumental in low variates of DBH activity and very likely such variates within the normal range of physiological variation. Moreover, it is suggested that the combined effects of MAO- and DBH-reduced efficiency on the metabolism of e.g. dopamine could be an essential pathogenic mechanism for the schizophrenic illness which is segregating in this population.     

The dominant diseases of modernized societies as omega-3 essential fatty acid deficiency syndrome: Substrate beriberi

About 1900, modern food selection and processing caused widespread $\text{epidemics}$ of the B vitamin deficiency diseases of beriberi and pellagra which, for genetic reasons, often expressed as different diseases ranging from bowel and heart disease to dermatoses and psychoses. But the B vitamins merely help convert essential fatty acids (EFA) into the prostaglandin (PG) tissue regulators and it now turns out that, through hydrogenation, milling and selection of w3-poor southern foods, we have also been systematically depleting, by as much as 90%, a newly discovered trace Nordic EFA (w3) of special importance to primates and sole precursor of the PG3(4) series, even as a concurrent fiber deficiency increases body demand for EFA. Since substrate EFA is processed by many B vitamin catalysts, an EFA deficiency will mimic a $\text{panh}$ypovitaminosis B, i.e., a mixture of $\text{substrate}$ beriberi and $\text{substrate}$ pellagra resembling vitamin beriberi and pellagra but exhibiting as even more diverse $\text{endemic}$ disease. This would consitute a second stage of the Modern Malnutrition and explain why some workers now hold the dominant diseases of modermized societies to be new, nutritionally based, pellagraform yet lipid-related and to range, once again, from heart disease to psychosis. It is an assumption that our dominant diseases are unrelated to each other or are merely revealed by our diagnostic acumen and therapeutic success; and that hydrogenating millions of tons of food oils annually, to destroy the rancidity producing w3-EFA, is safe for $\text{primates}$. Extensive beriberiform disease is reported here in 32 typical cases taken from medical practice which responds strikingly to linseed oil supplements (60% w3-EFA) in confirmation of identical results in Capuchins.     

What will studies of Fulani individuals naturally exposed to malaria teach us about protective immunity to malaria?

There are an estimated over 200 million yearly cases of malaria worldwide. Despite concerted international effort to combat the disease, it still causes approximately half a million deaths every year, the majority of which are young children with Plasmodium falciparum infection in sub-Saharan Africa. Successes are largely attributed to malaria prevention strategies, such as insecticide-treated mosquito nets and indoor spraying, as well as improved access to existing treatments. One important hurdle to new approaches for the treatment and prevention of malaria is our limited understanding of the biology of Plasmodium infection and its complex interaction with the immune system of its human host. Therefore, the elimination of malaria in Africa not only relies on existing tools to reduce malaria burden, but also requires fundamental research to develop innovative approaches. Here, we summarize our discoveries from investigations of ethnic groups of West Africa who have different susceptibility to malaria.     

'Very sore nights and days': the child's experience of illness in early modern England, c.1580-1720

Sick children were ubiquitous in early modern England, and yet they have received very little attention from historians. Taking the elusive perspective of the child, this article explores the physical, emotional, and spiritual experience of illness in England between approximately 1580 and 1720. What was it like being ill and suffering pain? How did the young respond emotionally to the anticipation of death? It is argued that children’s experiences were characterised by profound ambivalence: illness could be terrifying and distressing, but also a source of emotional and spiritual fulfilment and joy. This interpretation challenges the common assumption amongst medical historians that the experiences of early modern patients were utterly miserable. It also sheds light on children’s emotional feelings for their parents, a subject often overlooked in the historiography of childhood. The primary sources used in this article include diaries, autobiographies, letters, the biographies of pious children, printed possession cases, doctors’ casebooks, and theological treatises concerning the afterlife.     

Guidelines for the Validation and Use of Immunoassays for Determination of Introduced Proteins in Biotechnology Enhanced Crops and Derived Food Ingredients

Recent advancements in agricultural biotechnology have created a need for analytical techniques to determine introduced proteins in crops enhanced through modern biotechnology techniques. These proteins are expressed in plant tissues and may be present in food ingredients. Immunoassays are ideally suited for protein detection and may be used as both quantitative and threshold methods. Microplate ELISA and lateral flow devices are two of the most commonly used immunoassay formats for agricultural biotechnology applications. This paper provides general background information and a discussion of criteria for the validation and application of immunochemical methods to the analysis of proteins introduced into plants and food ingredients using biotechnology methods. It is the result of a collaborative effort of members of the Analytical Environmental Immunochemical Consortium. This collaborative effort represents the combined expertise of several organizations to reach consensus on establishing guidelines for the validation and use of immunoassays. Further, the paper offers developers and users a consistent approach to adopting the technology as well as aid in producing accurate and meaningful results.     

Ultracryotomy: development and application (with examples from the yeast cytology) (author's transl)

The preparation steps usually necessary for obtaining ultrathin frozen sections of biological material (chemical prefixation, enclosing, cryoprotective treatment, freezing, sectioning, and post-staining the sections for transmission electron microscopy) are submitted to a critical analysis. The application of cryo-ultramicrotomy, in particularly for cytochemical purposes, is reviewed. Fundamental considerations of chemical prefixation and poststaining are supported by examples from yeast cytology. Furthermore, the efficiency of the cryo-ultramicrotomy (electron optical resolution of ultrastructural details) is demonstrated on yeast cells and protoplasts.     

HLA in North Colombia Chimila Amerindians

HLA-A,-B,-C,-DRB1 and -DQB1 alleles have been studied in Chimila Amerindians from Sabana de San Angel (North Colombian Coast) by using high resolution molecular typing. A frequent extended haplotype was found:HLA-A*24:02-B*51:10-C*15:02-BRB1*04:07-DQB1*03:02 (28.7%) which has also been described in Amerinndian Mayos Mexican population (Mexico, California Gulf, Pacific Ocean). Other haplotypes had already been found in Amerindians from Mexico (Pacific and Atlantic Coast), Peru (highlands and Amazon Basin), Bolivia and North USA. A geographic pattern according to HLA allele or haplotype frequencies is lacking in Amerindians, as already known. Also, five new extended haplotypes were found in Chimila Amerindians. Their HLA-A*24:02 high frequencies characteristic is shared with aboriginal populations of Taiwan; also, HLA-C*01:02 high frequencies are found in New Zealand Maoris, New Caledonians and Kimberly Aborigines from Australia. Finally, this study may show a model of evolutionary factors acting and rising one HLA allele frequency (-A*24:02), but not in others that belong to the same or different HLA loci.     

The universal muscular chamber. A basic unit of the genitourinary, cardiovascular, gastro-intestinal, skeletal, cutaneous, respiratory and other systems, endocameral hypertension; and spasm, the key to their idiopathic diseases and arthropathies

Starting with the integument, we see many organs are contractile sacs or multiples thereof, which tubes or bags constitute the major part of the entire body. Recognition of this basic unit and its characteristics sheds new light, individually and collectively, on many disorders previously considered unrelated. Muscular tears and perforations develop in the walls of these chambers, being no way peculiar to those organs, wherein, hydrochloric acid occurs. So, it is not necessary to explain the absence of excessive acid from patients who exhibit holes in the gastric, uterine, aortic, duodenal, rectal, pulmonary, retina, and other walls. Muscle, not acid is the great common factor relating idiopathic disorders in the gastrointestinal tract to each other and to similar diseases in other systems. When the units are linked together, the lesions tend to appear as arthropathies, i.e. at the joints. Rephrasing common-place observations, frees us from conventional, conceptual cul-de-sacs. An observation is only as good as its interpretation, so all possibilities must be considered, otherwise, we will remain blinded by our misconceptions.