肿瘤抗原肽研究新进展

肿瘤抗原肽的寻找已成为当前肿瘤研究领域的一大热点。它包括癌基因蛋白抗原肽、致癌病毒基因抗原肽、肿瘤特异性抗原肽和组织特异性分化抗原肽等。这些抗原肽在与MHC类分子结合后 ,提呈到细胞表面被相应T细胞受体识别 ,从而激活细胞毒性T淋巴细胞的活性 ,使机体产生特异性抗肿瘤免疫应答。因而它们在肿瘤免疫接种和免疫治疗等方面有着广阔的应用前景。     

利用MHC-I/肽结合的预测与免疫学实验相结合的方法鉴定肿瘤抗原表位

目的:建立中国人群常见的人类白细胞抗原(HLA)分子限定的肿瘤抗原免疫表位的鉴定技术,为分离及鉴定肿瘤特异性T细胞以及克隆肿瘤抗原特异性T细胞受体基因提供实验基础。方法:以肿瘤/睾丸抗原NY-ESO、MAGE-A1和KK-LC-1作为模型,选用中国人群常见的HLA-I分型,利用主要组织相容性复合体(MHC)/肽预测软件对这3个抗原的MHC-I类表位进行预测。用PCR技术测定健康志愿者的HLA分型,以志愿者含有的中国人群常见HLA-A*11:01和HLA-B*46:01分型选择预测的抗原表位,设计合成KK-LC-1长肽,进行T细胞刺激实验,用ELISPOT和流式细胞仪分析产生干扰素γ(IFN-γ)的T细胞数量和CD137上调反应,以验证肽对T细胞特异性刺激的有效性。对其中一个反应最好的KK-LC-1长肽,开展短肽的验证。结果:(1)3个肿瘤/睾丸抗原在我国人群35种常见HLA分型中存在众多的强结合表位,在不同蛋白质的序列中分布不均匀;(2)KK-LC-1长肽可刺激T细胞产生T细胞活化反应,即释放IFN-γ和T细胞上调CD137分子,与预测的HLA分型基本吻合;(3)KK-LC-1短肽比长肽刺激效果更好,有更精确的抗原表位。结论:利用MHC/肽预测法并结合抗原特异性刺激实验可快速鉴定肿瘤抗原的T细胞表位;根据长肽的结果缩短肽段,可确定抗原表位。本研究为快速确定肿瘤抗原表位提供了新的途径。     

HLA—B27和强直性脊椎炎

HLA－B27与强直性脊椎炎呈强关联,但它在强直性脊椎炎中作用仍不祥,对HLA－B27分子的结构特征、抗原肽与B27亚型以及CTL细胞对抗原肽－B27复合体表位识别进行深入的研究,有助于了解其在强直性脊椎炎发病中的确切作用,本文拟就有关方面的研究进展作一综述。     

084 HLA-B27和强直性脊椎炎

HLA-B27与强直性脊椎炎呈强关联,但它在强直性脊椎炎中作用仍不详,对HLA-B27分子的结构特征、抗原肽与B27亚型以及CTL细胞对抗原肽-B27复合体表位识别进行深入的研究,有助于了解其在强直性脊椎炎发病中的确切作用,本文拟就有关方面的研究进展作一综述.     

B细胞受体(BCR)抗原特异性的研究进展

B细胞受体(BCR)是B细胞特异性识别和结合抗原产生适应性体液免疫应答的关键分子。B细胞分化过程中的基因重排和体细胞高频突变是形成BCR多样化的主要机制。BCR极大的多样性和独特的分子结构决定了抗原识别的多样性和特异性，形成了具有广泛抗原特异性的复杂B细胞库。因此，BCR抗原特异性信息是理解不同疾病适应性免疫特征的关键。随着B细胞相关研究技术的发展，如单细胞分选技术、高通量测序(HTS)技术、 B细胞受体与抗原特异性结合测序(LIBRA-seq)等，提高了将BCR序列与抗原特异性联系起来的能力。这有助于更好地理解体液免疫反应、探索疾病发病机制、监测疾病进展、设计疫苗、研发治疗性抗体和药物。我们主要总结了感染、疫苗接种、自身免疫性疾病及肿瘤的抗原特异性BCR相关研究，并以SLE为例对其自身抗体序列进行分析，其序列特征使自身抗原的识别成为可能。     

人食管癌TE-1细胞MHC-Ⅰ相关抗原肽的初步研究

目的寻找TE-1细胞MHC-Ⅰ类分子提呈的抗原肽。方法用弱酸洗涤法获得TE-1细胞表面与MHC-Ⅰ类分子结合的抗原肽，经SepPak-C18柱、Centrion-3超滤器和RP-HPLE去盐纯化后，用负载该抗原肽的树突状细胞刺激生成肿瘤特异性杀伤细胞，以Cr^51释放法测定CTL的杀伤活性。结果高效液相色谱图显示多个峰；负载抗原肽DC刺激的CTL比未负载抗原肽DC刺激的CTL对TE-1细胞杀伤率高，两者差异有统计学意义；负载抗原肽DC刺激的CTL对与抗原肽孵育的T2细胞比对未经与抗原肽孵育的T2细胞杀伤率高，两者差异有统计学意义。结论弱酸洗涤法可从TE-1细胞上获得有效的抗原肽；混合抗原肽中有可与HLA-A2分子结合并能激发CTL的抗原肽。     

CTL对同种异体靶细胞杀伤作用的实验研究

目的：研究特异性CTL杀伤的效应与HLA型别之间的关系。方法：用已知型别的EB病毒转化的B淋巴母样细胞（Epstein-Barr-transformed B lymphoblastoid cell line,EBV-LCL）与同种异体外周血单个核细胞（PBMC）共培养，激活同种异体抗原特异性细胞毒性T细胞（cytotoxicity T cell,CTL），然后利用同位素释放法观察CTL对HLA－I类表型不同的EBV－LCL的杀伤活性。结果：用EBV－LCL－1刺激自身PBMC－1所诱翌的CTL－1a，只能杀伤EBV－LCL－1，而不能杀伤HLA－I类表型不同的EBV－LCL－2；但用EBV－LCL－2刺激PBMC－1所诱导的CTL－1b，却能有效杀伤HLA－I类表型不同的EBV－LCL－2。结论：①MHC表型不同的免疫细胞之间可以发生相互作用；②TCR既不单独识别靶细胞表面的抗原肽，也不直接识别靶细胞表面的MHC分子（MHC特异性抗 原决定簇），而是识别MHC－抗原肽复合物的表达综合信息，后者可能是由MHC－抗原肽复合物表面的空间构象、电荷性质及其分布等信息所构成；③所谓CTL的特异性杀伤作用，是MHC－抗原肽复合物表面信息激活该信息性T细胞克隆的结果。     

MHC I类分子及与之结合的抗原肽

细胞内生性抗原需降解成抗原肽，由ＭＨＣ Ｉ类分子呈递给ＣＤ８＾＋细胞识别，从而诱导特异性细胞免疫应答。ＭＨＣ Ｉ类分子与抗原肽之间的结合表现出ＭＨＣ等位基因特异性，根据此特点可以预测抗原的Ｔ细胞识别表位，对于阐明自身免疫病和器官移植排斥反应的机理，指导人工合成肽疫苗用于病毒感染及肿瘤的免疫防治均具有重要意义。     

sHLA-A*0201-抗原肽四聚体的构建与功能检测

目的：构建有功能的sHLA-A＊0201-抗原肽四聚体，建立一套HLA Ⅰ类分子／抗原肽的四聚体制备技术。方法：利用基因工程及体外折叠技术构建sHLA-A＊0201-LMP2A426-434复合物分子，并在BirA酶的作用下生物素化。与荧光标记的亲和素衍生物以分子数4：1结合而形成HLA-A＊0201-LMP2A426-434四聚体。获得的四聚体对长期混合淋巴细胞培养中增殖的抗原特异性CTL进行检测，同时用细胞毒实验验证。结果：荧光标记的亲和素与生物素化的HLA-A＊0201-抗原肽复合物分子正确结合，制备的四聚体能够与长期混合淋巴细胞培养出的特异性T细胞结合。结论：从结构与功能上证实成功地获得有功能的HLA-A＊0201-抗原肽复合物四聚体。为进一步研究T细胞识别机制与功能奠定了基础，也为过程复杂的HLA-抗原肽四聚体制备提供了可行的免疫学监控方法。     

十肽表位HPV-16E711-20氨基酸置换修饰的研究

目的 采用氨基酸置换对人乳头状瘤病毒16型口抗原的人白细胞抗原A2分子限制性细胞毒性T细胞表位HPV-16E7 11-20进行修饰和鉴定。方法 运用量化模体方案对置换后的多肽与人白细胞抗原A2分子的结合系数进行比较，以分子模拟方法确定合成序列，采用标准Fmoc方案进行合成与纯化多肽以及标准^51 Cr释放试验检测特异性细胞毒性T细胞诱导活性。结果 修饰多肽符合人白细胞抗原A2分子限制性细胞毒性T细胞的表位要求，十肽YLLDLQPEVT具有特异性细胞毒性T细胞诱导活性。结论 修饰表位YLLDLQPEVT具有更好的结合力和较强的抗原性，可以代替原有序列E7 11-20（YMLDLQPEIT）作为人乳头状瘤病毒感染治疗性肽疫苗分子设计的新表位。     

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.     

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.