丙型肝炎病毒核酸疫苗的研究

从丙型肝炎病人的阳性血清中提取出丙型肝炎病毒ＲＮＡ，通过ＲＴ－ＰＣＲ的方法获得丙型肝炎病毒Ｃ区基因片段。并将此片段克隆到真核细胞表达载体ｐｃＤＮＡ３．１（＋），得到重组质粒ｐｃＤＮＡ－ＨＣＶ／Ｃ，再将其通过肌肉注射免疫ＢＡＬＢ／小鼠后，小鼠产生了抗ＨＣＶ／Ｃ区抗体。     

广西肝细胞癌与丙型肝炎的关系

用４种抗丙型肝炎病毒蛋白的单克隆抗体，免疫酶染色法直接检测肝细胞癌的病理组织中丙肝病毒蛋白。结果８／５２例（１５．４％），一至数种抗体阳性，阳性反应物主要定位于肝和癌细胞的胞浆中，值得注意的是本组几乎全部病例为乙肝，丙肝两种病毒混合感染，表明广西丙型肝炎的混合感染的比较高，乙型肝炎病毒（ＨＢＶ）加上黄曲霉毒素（ＡＦＢ１）或丙型肝炎病毒（ＨＣＶ）可能是广西肝细胞癌（ＨＣＣ）高发的原因。     

反显性突变Tat蛋白抑制人免疫缺陷病毒1型的复制

Ｔａｔ（Ｔｒａｎｓａｃｔｉｖａｔｏｒ）蛋白是人免疫缺陷病毒ＨＩＶ基因组编码的返式式激活因子。为了进一步研究Ｍ５Ｔａｔ和Ｍ６Ｔａｔ蛋白对ＨＩＶ病毒复制的抑制作用，将可被Ｔａｔ调控的启动子ＹＬ１５８替换基因治疗中常用的逆转录病毒载体ＬＮＣＸ中的ＣＭＶ启动子，构建了表达上述反显性Ｔａｔ蛋白的逆转录病毒载体，随后导入双向性包装细胞ＰＡ３１７细胞中，用产生出的复制缺陷病毒感染ＭＴ４Ｔ淋巴细胞。用ＨＩＶ－１病     

对丙型肝炎病毒具有潜在抑制作用的 L-苯丙氨酸衍生物前体的合成

丙型肝炎病毒（HCV ）感染与肝脏病变密切相关，是造成慢性肝炎、肝硬化及肝脏细胞肿瘤的主要原因之一．由于病毒自身的生物学特性，目前仍然没有有效的疫苗和治疗药物，研究开发高效的抗病毒药物具有重要意义．Reddy和他的小组设计合成了一系列 N，N-二取代的苯丙氨酸类似物，并证明了具有该种结构的化合物可对肝炎病毒聚合酶具有很好的抑制作用，以 L-苯丙氨酸为起始原料，经过酯化成氨基酸甲酯、与不同取代基的芳香醛缩合成希夫碱、将亚胺结构还原、酸化脱酯4步反应合成了8种对丙型肝炎病毒具有潜在抑制作用的此类化合物的前体，最高总产率达到77．4％．测定产物熔点，通过核磁氢谱对目标产物的结构进行表征，结果表明实现了目标化合物的合成．     

乳酸菌饲料添加剂对小鼠免疫功能的影响

观察了ＬＡＢＦＡ对小鼠免疫功能的影响．结果表明：ｉｇＬＡＢＦＡ１．２５、２．５Ｏ和５．００×１０７ＣＦＵ／只，每天１次，连续１２ｄ，能激活腹腔巨噬细胞吞噬活性，促进吞噬作用，对抗由Ｈｙｄ导致腹腔巨噬细胞吞噬功能的降低；能提高血清溶血素及ＩｇＭ水平，促进抗体生成；能促进淋巴细胞转化，提高由ＣＲＢＣ诱发的ＴＤＨ反应；可增加胸腺和脾脏重量．这些结果说明ＬＡＢＦＡ对小鼠免疫功能有一定的促进作用．     

庚型肝炎病毒研究进展

一、GB肝炎因子的研究历史 1967年,Deinhardt等在研究肝炎病毒的传播途径和动物模型时,用5份血清分别给绒猴注射时,其中2份血清诱发典型肝炎。这两份血清一份为混合血清,另一份来自一名34岁患黄疸3天后的临床医生,该医生名字的英文缩写为GB,故将该病人血清中的致病因子称为GB因子。绒猴感染GB因子发病后其血清仍可感染其它绒猴。通过比较研究发现,GB因子不是甲肝病毒(HAV),也非乙型肝炎病毒(HBV),而是另一类病毒,当时归为非甲非乙型肝炎病毒(NANBV)范畴,也有人称其为丙型肝炎病毒     

FK506对人外周血T淋巴细胞CD28分子表达的体外…

应用淋巴本外化模型和流式细胞分析技术，观察异体角膜体外对人外周血Ｔ淋巴细胞ＣＤ２８分子表达的调节和ＦＫ５０６对Ｔ淋巴细胞ＣＤ２８表达的抑制作用，结果显示：空白对照组Ｔ淋巴细胞ＣＤ２８表达为２１．３％；受异体角膜或ＰＤＢ刺激后，Ｔ淋巴细胞ＣＤ２８表达分别为５２．４％和８１．４％，同时加入ＦＫ５０６的实验组，Ｔ淋巴细胞ＣＤ２８表达分别为２４．５％和４１．９％，结果表明异体角膜体外能够刺激Ｔ淋巴细胞ＣＤ     

细胞与病毒中的RNA依赖的RNA聚合酶

就细胞与病毒的RdRP的结构及其功能，以丙型肝炎病毒的NS5B蛋白和RNA干扰系统为例，介绍了RdRP的研究进展．     

FK506对人外周血T淋巴细胞CD_(28)分子表达的体外抑制作用

应用淋巴细胞体外活化模型和流式细胞分析技术，观察异体角膜体外对人外周血Ｔ淋巴细胞ＣＤ２８分子表达的调节和ＦＫ５０６对Ｔ淋巴细胞ＣＤ２８表达的抑制作用．结果显示：空白对照组Ｔ淋巴细胞ＣＤ２８表达为２１．３％；受异体角膜或ＰＤＢ刺激后，Ｔ淋巴细胞ＣＤ２８表达分别为５２．４％和８１．４％，同时加入ＦＫ５０６的实验组，Ｔ淋巴细胞ＣＤ２８表达分别为２４．５％和４１．９％·结果表明异体角膜体外能够刺激Ｔ淋巴细胞ＣＤ２８分子表达，ＦＫ５０６体外能够明显抑制Ｔ淋巴细胞ＣＤ２８分子表达和活化．     

HCV感染的动物模型研究进展

丙型肝炎（HC）是由丙型肝炎病毒（HCV：一种正链RNA病毒[1]）引起的一种传染性疾病，该病毒的流行呈世界性分布。HC曾一度归入NANBH中，到1989年Choo[1]等，从实验感染的黑猩猩血液中成功地分离出丙肝病原体并成功克隆出其全部核苷酸，才确证了HCV为NANBH的主要病原体的证据，同年在日本召开的国际病毒性肝炎会议，正式将病毒性肝炎分为甲、乙、丙、丁、戊五型；并将相应的病毒命名为HAV、HBV、HCB、HDV、HEV。近年来的研究证实病毒性肝炎还存在另外两型即已型肝炎（HF）和唐型肝炎（HG）。HC病原体分离出来后，对HC的…     

绒白乳菇中提取的乳菇菌素-D免疫抑制作用的研究

研究从绒白乳菇菌中分离出的一种新型倍半萜类化合物-乳菇菌素D的免疫抑制作用及机制。运用体外培养法取BALB/C小鼠脾细胞,研究乳菇菌素D对静止脾细胞、Con A(刀豆蛋白A)和LPS(脂多糖)刺激后的脾细胞(含B淋巴细胞和T淋巴细胞)增殖的抑制作用;运用MTT法经酶标仪检测吸光度,并计算刺激指数(stimulating index,SI);体外培养小鼠脾细胞,运用ELISA检测法观察乳菇菌素D对脾脏中T淋巴细胞分泌IL-2的抑制作用。结果显示该化合物对静止的脾细胞无抑制作用;对Con A和LPS刺激后的脾细胞增殖有显著抑制作用(P0.05),最大浓度药物组刺激指数SI低至0.28;高浓度的乳菇菌素D对Con A刺激的小鼠脾细胞产生的IL-2,具有显著抑制作用(P0.05)。结果显示乳菇菌素D通过抑制抗原刺激后的T淋巴细胞和B淋巴细胞的增殖、抑制其分泌细胞因子,从而发挥免疫抑制作用。研究结果将为开发以乳菇菌素D为母体、经结构修饰、安全低毒的免疫抑制剂提供理论基础。     

乙型肝炎病毒蛋白及COX-2在乙肝相关性肝细胞癌发展与转移中的作用机制

目的探讨乙型肝炎病毒蛋白及环氧合酶-2(COX-2)在乙肝相关性肝细胞癌发展与转移中的作用机制.方法取42例慢性乙肝患者行穿刺活检时乙型肝炎病毒ccc DNA为阳性乙肝相关性肝细胞癌组织;另取同期手术切除的11例ccc DNA为阴性的非乙型肝炎相关性肝癌组织.免疫组化法检测乙型肝炎病毒X蛋白、COX-2、CD34的表达水平,Werdner法计算微血管密度;分析上述因子与乙肝相关性肝细胞癌组织微血管生成的相关性.RT-PCR和Western blot检测人肝癌细胞系(HepG2)和稳定转染乙型肝炎病毒X蛋白(HepG2-X)细胞中COX-2mRNA和蛋白表达情况;ELISA法检测细胞上清液中PGE2表达水平和不同浓度COX-2抑制剂塞来昔布作用后PGE2水平.结果乙型肝炎病毒X蛋白阳性表达组织中COX-2阳性率明显高于乙型肝炎病毒X蛋白阴性表达组织和非乙型肝炎相关性肝癌组织(P0.01).乙型肝炎病毒X蛋白阳性表达组织中早期癌症微血管密度明显低于进展期癌症组织,乙型肝炎病毒X蛋白阴性表达组织中微血管密度明显低于阳性表达组织(P0.01);COX-2阳性表达组织中微血管密度明显高于COX-2阴性表达组织(P0.01);非乙型肝炎相关性肝癌组织中微血管密度明显低于乙型肝炎病毒X蛋白、COX-2阳性表达组织(P0.01),与乙型肝炎病毒X蛋白阴性表达组织和COX-2阴性表达组织之间差异无统计学意义(P0.05);乙型肝炎病毒X蛋白、COX-2在乙型肝炎相关性人肝细胞癌组织微血管生成呈正相关.HepG2-X细胞中COX-2 mRNA和蛋白表达水平明显高于空载体对照HepG2细胞,并且细胞培养上清液中PGE2水平明显增加;与HepG2细胞相比,塞来昔布对HepG2-X细胞分泌PGE2具有更强的抑制作用.结论乙型肝炎病毒X蛋白、COX-2在乙肝相关性肝细胞癌组织中高表达,促进了癌组织微血管生成;乙型肝炎病毒X蛋白可通过COX-2/PEG2信号通路促进了肝癌的发生和发展.     

Similarities between interleukin-2 receptor number and affinity on activated B and T lymphocytes

Interleukin-2 (IL-2) is a T-cell-derived polypeptide hormone of 133 amino acids which exerts its growth-promoting activity via a surface receptor. Originally, IL-2 was believed to be a unique growth factor for activated T cells; more recent studies, however, have demonstrated that certain B-cell tumours as well as normal activated B lymphocytes express a surface molecule which is recognized by monoclonal antibodies directed against the IL-2 receptor. Furthermore, we and others have shown recently that activated B cells proliferate in response to either immunoaffinity-purified or recombinant IL-2. These controversial findings prompted us to undertake a detailed quantitative comparison of IL-2 receptor expression on activated B and T cells. We show here, using biosynthetically labelled IL-2(3H-IL-2) and anti-IL-2 receptor antibody (3H-PC61) that activated B and T cells express both high-affinity (apparent dissociation constant, Kd approximately 20 pM) and low-affinity (Kd approximately 1,000 pM) IL-2 receptors. Binding of IL-2 to both classes of receptor is inhibited by the monoclonal anti-IL-2 receptor antibody PC61. B blasts express half as many total IL-2 binding sites or PC61 binding sites as T blasts, and the ratio of the number of low- to high-affinity receptors for each cell type is approximately 10:1. Immunoprecipitation analysis of surface-labelled blasts indicates that B and T cells have IL-2 receptors of similar relative molecular mass. Taken together, these data suggest strongly that IL-2 can act as a growth hormone for both B and T lymphocytes.     

我国戊型肝炎全病毒抗原抗HEV·IgM酶联诊断盒的临床应用

我国戊型肝炎全病毒抗原抗ＨＥＶ·ＩｇＭ酶联诊断盒的临床应用廖绵初，干侣仙，黄如统，李晓萸，李德荣（厦门大学抗癌研究中心）（军事医学科学院五所）赵炳华（浙江医科大学第一附属医院）从１９８９年正式确立戊型肝炎ＨｅｐａｔｉｔｉｓＥ以来，许多国家都研制其诊断...     

Human interleukin-2 promotes proliferation of activated B cells via surface receptors similar to those of activated T cells

Human interleukin-2 (IL-2) is a glycoprotein of relative molecular mass (Mr) 15,000, which is released by T lymphocytes on stimulation with antigen or mitogen and functions as a T-cell growth factor (TCGF) by inducing proliferation of activated T cells. It is generally accepted that resting or activated B cells do not respond directly to IL-2 but require for their proliferation other T-cell-derived lymphokines usually referred to as B-cell growth factors (BCGFs). Recently, however, a monoclonal antibody reacting with the IL-2 receptor molecules expressed by activated T cells (anti-Tac) was shown to react also with certain B tumour cells; in addition, murine B cells proliferate in response to pure human IL-2. We now show that recombinant IL-2, derived from Escherichia coli expressing the human gene, is able to promote strong proliferation of human B cells activated with protein-A-rich Staphylococcus aureus Cowans strain I. Moreover, we demonstrate that the anti-Tac antibody also reacts with S. aureus-activated normal B cells and inhibits sharply the proliferative response of such cells to IL-2. Finally, immunoprecipitation experiments reveal that anti-Tac defines similar molecules on activated T and B cells.     

Mitotic EBNA-positive lymphocytes in peripheral blood during infectious mononucleosis

Infectious mononucleosis (IM) is usually a benign lymphoproliferative disease caused by Epstein-Barr virus (EBV). Although EBV induces a state of continuous proliferation in infected B lymphocytes in vitro, the most prominent lymphoproliferation during IM is of activated, or atypical, T lymphocytes presumably responding to the virus or virus-infected cells. However, EBV genome-carrying cells are known to be circulating during IM, as cultured peripheral blood leukocytes from patients with the disease give rise to continuous lymphoblastoid cell lines, each cell of which contains the EBV genome and expresses the EBV determined nuclear antigen (EBNA). The proposal that EBV-infected cells in IM blood are not endowed with enhanced growth potential but are merely latently infected is supported by demonstrations that cells infected in vivo enter a viral replicative cycle when placed in vitro and that most cell lines derived from cultured lymphocytes of IM patients are infected by virus released in vitro. However the cells could also be capable of proliferation in vivo, since virus production and transformation are not mutually exclusive properties of EBV-transformed cells. Recently, EBNA has been detected in a very small fraction of peripheral blood lymphocytes of IM patients after T cells were first removed and this has been interpreted to indicate that cell transformation occurs in vivo during IM. The isolation of colonies of EBNA-positive cells from IM blood leukocytes cultures in soft agar suggests that at least some infected cells are capable of direct outgrowth into transformed cells. We report here direct evidence that circulating EBV-infected cells exhibit increased growth properties during IM.     

Selective killing of hepatitis B envelope antigen-specific B cells by class I-restricted, exogenous antigen-specific T lymphocytes

Specific B lymphocytes can act as very efficient antigen-presenting cells. They bind antigen with high affinity via their immunoglobulin receptors, process it through the class II major histocompatibility complex (MHC) pathway, and present its fragments to class II-restricted T lymphocytes. In general, exogenous antigens and noninfectious viral particles enter the class II pathway and are selectively associated with class II MHC molecules. The presentation of an exogenous antigen in association with class I molecules has been reported for only a few antigens, including the hepatitis B envelope antigen (HBenvAg). Here we demonstrate that antigen-specific B cells can efficiently deliver HBenvAg to the class I pathway, presenting its fragments to class I-restricted cytotoxic T lymphocytes (CTLs) which kill the specific B cells. This could represent a mechanism of suppression of neutralizing anti-hepatitis B virus (HBV) antibody response, a phenomenon that accompanies the development of the chronic HBV-carrier state.     

恩诺沙星对实验小鼠细胞免疫的动态影响

目的研究恩诺沙星对实验小鼠免疫细胞的影响。方法24只SPF级小鼠随机分成恩诺沙星组与正常组。实验分3周进行,每组每周各杀4只,采用酸性α-醋酸萘酯酶（ANAE）方法检测T、B淋巴细胞的动态比值变化。结果喂药1周小鼠的T、B淋巴细胞比值与正常组差异极显著（P〈0.01）,提示该药对T、B淋巴细胞有影响;而喂药第2、3周小鼠的T、B淋巴细胞比值的影响差异不显著（P〉0.05）。结论恩诺沙星在短期内对实验小鼠免疫细胞有影响,尤其是T淋巴细胞。     

Precursor and effector phenotypes of activated human T lymphocytes

In mice, thymus-derived lymphocytes are differentiated into functional subclasses by their cell surface antigens. The Ly 1 determinants are present on T cells with a helper function, whereas Ly 2 and Ly 3 antigens are expressed on the surface of lymphocytes with suppressor or cytotoxic functions. In man also, T-cell subsets have been identified using allo- and heteroimmune sera and, more recently, using monoclonal antibodies, which seem to identify helper and suppressor or cytotoxic subpopulations. The major histocompatibility system (MHS)-encoded Ia antigens belong to several polymorphic families of membrane associated glycoproteins originally found on B lymphocytes; however, they have also been shown to be markers for suppressor T cells in mice. Recent studies have shown that in both mouse and man, T cells activated by a mixed lymphocyte reaction or by mitogens become Ia+. Furthermore, some human T lymphoid cells, either freshly isolated from peripheral blood or after in vitro activation by lectins or alloantigens, possess suppressor properties. We report here the phenotype of a T suppressor-cell subpopulation which was induced in long-term culture of lymphoid cells after activation with phytohaemagglutinin (PHA). Our results suggest that a subset of T cells was progressively expanded over a period of 8 days in culture and that, with the expression on the surface of these cells of 'Ia-like' antigens, they acquired the capacity to suppress the proliferative response of syngeneic or allogeneic lymphocytes to alloantigens or mitogens.     

C_2-神经酰胺对小鼠T淋巴细胞体外活化和增殖的影响

目的:研究C2-神经酰胺(C2-cer)对小鼠T细胞体外活化和增殖的影响,并对其免疫调节作用进行初步探讨.方法:分离小鼠淋巴结细胞,加入刀豆蛋白A(ConA)或佛波醇酯(PMA)和离子霉索(Ion)进行刺激,以不同终浓度的C2-cer与T细胞共培养.流式细胞术结合双色荧光抗体染色,检测T细胞早期活化抗原CD69的表达;用羧基荧光素双醋酸盐琥珀酰(CFDA-SE)染色结合流式细胞术,并用ModFit软件分析T细胞增殖相关指数.结果:终浓度为25、50和75 μmol/L的C2-cer可以显著抑制ConA诱导的T细胞CD69的表达,由ConA组的(60.13±1.18)%,分别降低为(54.56±1.14)%、(48.73±1.26)%和(27.09±1.07)%(P＜0.01);CFDA-SE染色结果显示,各浓度C2-cer对ConA诱导的小鼠淋巴细胞增殖,具有明显的抑制作用,增殖指数(PI)由ConA组的(1.81±0.25),分别降低为(1.46±0.01)、(1.25±0.04)和(1.18±0.03)(P＜0.05);各浓度C2-cer均可明显抑制PMA+Ion诱导的小鼠淋巴细胞增殖,增殖指数(PI)由PMA+Ion组的(1.47±0.01),分别降低为(1.27±0.01)、(1.11±0.01)和(1.05±0.01)(P＜0.05).结论:C2-cer能有效地抑制小鼠T细胞的体外活化和增殖.