29 人外周血树状突细胞抗肿瘤作用的探讨

<正>有关树状突细胞(Dendritic Cell,DC)在肿瘤免疫中作用的研究,多限于观察肿瘤局部DC的变化,而对DC抗肿瘤作用的探讨尚属起步.本研究采用多因素不同水平的全面试验设计,进行DC的体外抗肿瘤实验.首先应用新的三步分离法分离、纯化,获得纯度为60～70%的DC;又在用重组IL-2诱导淋巴因子激活的杀伤细胞(LAK 细胞)的基础上,以中性红摄入比色法检测细胞毒活性,观察了体外DC对LAK细胞杀伤H_7402肿瘤细胞活性的影响,也观察了形态学变化.结果:活细胞观察发现DC联合LAK作用组残存的活肿瘤细胞比单LAK作用组明显减少,HE染色见肿瘤细胞呈不同程度的坏死状态.免疫细胞化学染色显示S-100蛋白阳性的DC与LAK细胞形成花环,DC、LAK和肿瘤细胞三者形成细胞簇.扫描电镜下可见DC借助突起与肿瘤细胞相连,也与LAK细胞接触,三者形成细胞簇.细胞毒活性检测所得各孔光密度OD值经方差分析和参数估计,表明LAK细胞体外杀伤H_(7402)肿瘤细胞的活性随效靶比增加而增强,DC能协同LAK细胞的活性,上调其杀伤肿瘤的作用(P<0.01),并以中等剂量的DC上调速度最明显,加入IL-2后,DC的调节作用更有上升.提示人外周血DC在细胞免疫抗肿瘤过程中起重要使用.     

Hsp70-肿瘤抗原肽复合物修饰的DC疫苗体内外特异性抗瘤作用

目的 :探讨树突状细胞 (DC)及Hsp70 肿瘤抗原肽复合物修饰后的体内外特异性抗瘤作用。方法 :采用一定的生化技术 ,从H2 2肝癌细胞中提取肿瘤抗原肽 ,并在体外与Hsp70进行结合 ;采用细胞培养技术 ,培养rmGM CSF、rmIL 4诱导的小鼠骨髓细胞 ,体外获取大量的DC ,后者经Hsp70 肿瘤抗原肽复合物修饰后刺激小鼠脾淋巴细胞 ,通过MTT法进行检测淋巴细胞的激活 ;收集上述刺激传代培养的淋巴细胞 ,检测其对H2 2瘤细胞和艾氏腹水癌细胞的杀伤功能 ;采用H2 2瘤细胞肌肉接种和H2 2瘤细胞、艾氏腹水癌细胞腹腔接种 ,对接种小鼠给予经体外修饰的DC回输 ,观察其抑制肿瘤的效果。结果 :Hsp70 肿瘤抗原肽复合物可使DC成熟 ,大量分泌IL 12、TNF α、IL 1β等细胞因子 ,并能够使DC激活小鼠脾淋巴细胞 ;激活后传代培养的淋巴细胞对H2 2瘤细胞能够特异性地杀伤 ,而对艾氏腹水癌细胞无效 ;经Hsp 70 肿瘤抗原肽复合物修饰后的DC可作为一种有效的瘤苗 ,体内能特异性地抑制小鼠H2 2肿瘤生长。结论 :Hsp70 肿瘤抗原肽复合物能够很好地修饰体外诱导获取的DC ,使后者成为一种有效的瘤苗 ,体外能够特异性地激活淋巴细胞 ,体内有效地抑制肿瘤生长     

树突状细胞激活的肿瘤浸润淋巴细胞对荷瘤小鼠免疫功能影响的研究

目的:探讨H22细胞全细胞性抗原致敏的DC激活的TIL体外抗小鼠肝癌活性;并将H22细胞全细胞性抗原致敏的DC激活的TIL(H22-DC-TIL)过继免疫荷瘤小鼠,研究其对荷瘤小鼠免疫功能的影响。方法:从小鼠四肢长骨骨髓中获取DC,应用粒/巨噬细胞集落刺激因子(GM-CSF)、白介素-4(IL-4)和肿瘤全细胞性抗原致敏DC,然后用DC激活TIL,观察TIL在体外对H22细胞、Hepal-6细胞和B16细胞的杀伤活性;检测应用H22-DC-TIL后荷瘤小鼠的脾淋巴细胞的NK、LAK、CTL活性及血清TNF活性,并与对照组相比较。结果:(1)H22-DC-TIL具有很强的对H22细胞杀伤活性(杀伤率为71.31%±3.11%),明显高于其对Hepal-6和B16细胞的杀伤活性(杀伤率分别为50.11%±3.03%和30.31%±2.89%);也明显高于未经DC激活的TIL、H22-DC-小鼠脾淋巴细胞和未经DC激活的小鼠脾淋巴细胞对H22细胞杀伤活性(杀伤率分别为49.80%±3.21%、48.76%±3.60%和19.23%±2.71%)和对Hepal-6细胞杀伤活性(杀伤率分别为39.40%±3.21%、38.62%±2.87%和18.73%±2.40%)以及对B16细胞杀伤活性(杀伤率分别为26.38%±2.51%、25.82%±2.70%和18.34%±3.01%),同时B16-DC-TIL(TIL来源于H22瘤体)也可诱导相对较低的对B16细胞的特异性细胞杀伤活性。(2)H22-DC-TIL可明显诱导提高荷瘤小鼠脾淋巴细胞NK、LAK和CTL活性(活性分别为30.43%±1.35%、31.40%±1.80%和35.30%±1.20%),并可检测到血清TNF水平明显上升[血清TNF为(40.41±1.85)U/m l],它们均达正常对照组水平,与未经DC激活的TIL组、H22-DC-小鼠脾淋巴细胞组、未经DC激活的小鼠脾淋巴细胞组、生理盐水组分别对应比较,差异均有显著性(P<0.01)。结论:(1)H22-DC-TIL可产生很强的体外针对H22细胞的特异性杀伤活性。(2)H22-DC-TIL可明显诱导提高荷瘤小鼠特异性抗肿瘤免疫反应。     

灵芝孢子粉对荷HAC肝癌小鼠抗肿瘤的实验性研究

本文通过荧光细胞检测技术、体外细胞毒试验以及肿瘤抑瘤率的测定 ,观察了灵芝孢子粉对荷HAC小鼠T细胞表面分化抗原 ,体外细胞杀伤功能以及肿瘤抑瘤率的影响。结果显示灵芝孢子粉治疗组中总T细胞的百分率 (6 3 4% )高于对照组 (5 6 3% ) ,其中对总T细胞、T辅助细胞的上调较为明显。在对HAC、YAC 1和P815肿瘤细胞的杀伤活性中灵芝孢子粉组的杀伤活性分别为 2 7 3%、 2 3 4%和 2 0 0 % ,高于对照组。灵芝孢子粉组的总抑瘤率为 42 2 %。结果表明灵芝孢子粉是一种能激活和提高特异性 (CTL )和非特异性杀伤细胞 (NK、LAK )的抗肿瘤作用 ,抑制肿瘤细胞的增殖以及调控机体免疫功能的中药。     

宫颈癌单克隆抗体AU14-1介导效应细胞对靶细胞的细胞毒

目的：研究单克隆抗体AU14-1介导的细胞毒作用对小鼠宫颈癌（U14）细胞的杀伤效果。 方法： 以小鼠淋巴因子激活的杀伤细胞（LAK）、脾淋巴细胞及巨噬细胞为效应细胞，应用MTT比色法测试在单克隆抗体AU14-1介导下对小鼠宫颈癌（U14）细胞的体外细胞毒作用。 结果： 3种效应细胞对于经单克隆抗体AU14-1处理的肿瘤细胞的杀伤率显著高于未经AU14-1处理的肿瘤细胞（P＜0.01）；以LAK的ADCC作用最强，巨噬细胞次之，脾淋巴细胞最弱（P＜0.01）。 结论： 单克隆抗体AU14-1能通过抗体依赖细胞介导细胞毒（ADCC）发挥淋巴样细胞、巨噬细胞及LAK细胞对靶肿瘤细胞的杀伤作用；提示特异性宫颈癌单克隆抗体AU14-1介导的ADCC在治疗宫颈癌中可能是一种有潜在价值的方法。     

HBV与HCV融合基因免疫的抗肿瘤作用研究

目的 :观察基因疫苗SpcDNA3.1、CpcDNA3.1及融合基因疫苗CSpcDNA3.1诱导BALB c小鼠 (H 2 d)的特异性细胞免疫应答及其对稳定表达HBsAg和HCcAg的小鼠肥大细胞瘤P815细胞 (H 2 d)接种动物后成瘤性的影响。方法 :3种重组质粒SpcDNA3.1、CpcDNA3 1和CSpcDNA3 1分别肌注免疫小鼠 ,3w后背部皮下接种质粒CSpcDNA3 1转染的P815细胞 ,观察小鼠成瘤和存活时间。LDH法检测免疫小鼠的脾淋巴细胞CTL活性。结果 :融合质粒CSpcDNA3 1可显著抑制肿瘤出现和生长 ,小鼠生存时间明显延长 ,生存率提高 ,CSpcDNA3 1免疫的小鼠脾淋巴细胞体外对转染的P815肿瘤细胞有明显的杀伤作用。结论 :融合基因疫苗CSpcDNA3 1免疫的小鼠能诱发特异性抗肿瘤细胞免疫。     

猪苓多糖、硒及红细胞对LAK细胞杀伤活性的影响

目的　探讨猪苓多糖 (PUPS)、微量元素硒 (Se)及红细胞 (RBC)对LAK细胞杀伤活性的影响。方法　分离正常人外周血单个核细胞 (PBMC)在体外分别与不同浓度的PUPS、亚硒酸钠及红细胞单独或与IL 2协同诱导 ,以MTT法检测LAK细胞活性 ,用ELISA检测培养上清中TNF α和IFN γ含量。结果　不同浓度的PUPS和Se能单独诱导LAK细胞 ,与IL 2协同诱导的LAK细胞活性明显提高 ,细胞分泌的TNF和IFN浓度增高。RBC对LAK细胞的杀伤活性有明显地增强作用 ,并呈浓度依赖关系。结论　PUPS、Se及RBC对LAK细胞杀伤性有明显增强作用 ,细胞杀伤活性的提高可能与内源性TNF和IFN的产生增加有关。本研究为PUPS、Se及RBC与IL 2协同诱导细胞的研究和临床应用提供了理论依据。     

联合应用IL-2及IL-4基因转染瘤苗对实验性肺转移肿瘤的治疗作用

将IL—2基因转染的高分泌IL—2的B16黑色素瘤细胞株(B2)及IL—4基因转染的高分泌IL—4的B16黑色素瘤细胞株(B4)制备成新型瘤苗,用以治疗实验性肺转移荷瘤小鼠,并辅以低剂量环磷酰胺,结果表明,荷瘤小鼠存活期明显延长;肺部转移结节数明显减少;两种瘤苗联合治疗后的疗效较单独使用明显,当辅以低剂量环磷酰胺时治疗效果更加明显。体内免疫功能检测表明,经两种瘤苗联合治疗的荷瘤小鼠脾细胞NK及CTL杀伤活性升高得更加明显,但对LAK活性及腹腔巨噬细胞杀伤活性无明显协同增强作用;脾细胞经诱导后产生的细胞因子中,IL—2含量有所升高。     

人LAK细胞的体外抗肿瘤作用

用重组IL-2(rIL-2)以及部分纯化的IL-2(PPIL-2)体外激活人外周血单个核细胞.(PBM),使之形成LAK细胞,然后借助于~(51)Cr释放实验,研究了正常人和肿瘤病人的LAk细胞对传代的肿瘤细胞系和新鲜实体瘤细胞的杀伤能力。实验结果表明:1.二种来源的LAK细胞均能明显杀伤传代的肿瘤细胞系,包括NK敏感的K562细胞和NK抵抗的Daudi细胞。2.采用数种新鲜实体瘤细胞作靶,二种来源的LAg细胞同样具有明显的广谱杀伤力,这证实该群杀伤细胞确系LAK细胞。3.不同来源的实体瘤细胞对LAK细胞的杀伤敏感性不同,表现为杀伤程度上的差异,这似乎提示某些肿瘤对LAK细胞杀伤存在抗性。     

接种CEA重组痘苗病毒对小鼠CEA^＋肿瘤抑制作用的机制探讨

目的 探讨CEA重组痘苗病毒（rV-CEA)治疗CEA^ 肿瘤的机制。方法 以我室构建的rV_CEA,腹腔接种供体C57/BL小鼠,取其脾细胞及腹腔巨噬细胞（Mφ）,分别过继转移给荷CEA^ -HePa肝癌细胞的C57/BL小鼠,检测该公共体小鼠脾细胞、腹腔Mφ及相应受体的脾细胞体外杀瘤细胞的效应,结果 接种rV-CEA的供体小鼠的脾细胞及腹腔Mφ过继免疫给受体小鼠,具有明显抑制受体CEA阳性肿瘤生长的作用,体外实验表明,该供体脾细胞及接种了供体Mφ的受体脾细胞对同一靶细胞的杀伤活性明显增强,但供体Mφ体外的细胞毒活性无明显增加,结论 rX-CEA对CEA^ 肿瘤的抑制作用,可能主要通过CEA特异性免疫反应激活T细胞而实现。Mφ作为抗原提呈细胞可通过激活T细胞而杀伤肿瘤细胞,具体机制值得进一步研究。     

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.     

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.     

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.     

'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.     

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.     

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.     

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.     

Der Einfluß von Nahrungsmitteln und Rauchen auf die klinisch-chemische Diagnostik von Phäochromozytom,Neuroblastom und Karzinoid-Syndrom

Zusammenfassung Der Einfluß von verschiedenen Nahrungsmitteln auf Methoden zur Bestimmung von Adrenalin (AD), Noradrenalin (NA), Vanillinmandelsäure (VMS), Metanephrinen (MN), Homovanillinsäure (HVS) und 5-Hydroxyindolessigsäure (5-HIE) im 24 h-Harn zur Diagnose des Phäochromozytoms bzw. Karzinoid-Syndroms wurde untersucht. Die in die Untersuchung einbezogenen Nahrungsmittel waren: Tee, Kaffee, Mandeln, Ananas, Käse, Walnüsse, Vanillepudding, Bananen, Tomaten und Milchschokolade. Außerdem wurde der Einfluß des Zigarettenrauchens auf die Bestimmung von AD, NA, VMS und MN untersucht.Walnüsse führten zu einer starken Erhöhung der 5-HIE-Ausscheidung. Bananen erhöhten die Ausscheidung von AD, NA, VMS, MN und 5-HIE. Kaffee und Ananas bewirkten eine geringe Zunahme der MN-Werte. Rauchen von 20–30 Zigaretten/Tag beeinflußte keine der vier Variablen.Wenn die beschriebenen Methoden benutzt werden, sollte lediglich auf den Verzehr von Bananen und Walnüssen vor und während der Harnsammelperioden verzichtet werden, da die oberen Normgrenzen im Harn überschritten werden könnten. Ein Verzicht auf Kaffee und Ananas in normalen Mengen ist nicht erforderlich. Es besteht kein Anlaß, weiterhin die bisherigen umfangreichen Restriktionen der übrigen Nahrungsmittel beizubehalten.