抗基因工程人rIFN-γ杂交瘤细胞株的建立

用杂交瘤技术将基因工程人γ－干扰素（ｒＩＦＮ－γ）免疫的Ｂ淋巴细胞与ｓｐ２／０鼠骨髓瘤细胞融合，建立了 ３株抗 ｒＩＦＮ－γ杂交瘤细胞株。这些细胞株经两年多的传代培养仍保持稳定分泌抗体的能力，并与ｒＩＦＮ－γ及新型γ－干扰素呈特异性反应，可用于亲和层析纯化γ－干扰素．     

Rapid cloning of high-affinity human monoclonal antibodies against influenza virus

Pre-existing neutralizing antibody provides the first line of defence against pathogens in general. For influenza virus, annual vaccinations are given to maintain protective levels of antibody against the currently circulating strains. Here we report that after booster vaccination there was a rapid and robust influenza-specific IgG+ antibody-secreting plasma cell (ASC) response that peaked at approximately day 7 and accounted for up to 6% of peripheral blood B cells. These ASCs could be distinguished from influenza-specific IgG+ memory B cells that peaked 14-21 days after vaccination and averaged 1% of all B cells. Importantly, as much as 80% of ASCs purified at the peak of the response were influenza specific. This ASC response was characterized by a highly restricted B-cell receptor (BCR) repertoire that in some donors was dominated by only a few B-cell clones. This pauci-clonal response, however, showed extensive intraclonal diversification from accumulated somatic mutations. We used the immunoglobulin variable regions isolated from sorted single ASCs to produce over 50 human monoclonal antibodies (mAbs) that bound to the three influenza vaccine strains with high affinity. This strategy demonstrates that we can generate multiple high-affinity mAbs from humans within a month after vaccination. The panel of influenza-virus-specific human mAbs allowed us to address the issue of original antigenic sin (OAS): the phenomenon where the induced antibody shows higher affinity to a previously encountered influenza virus strain compared with the virus strain present in the vaccine. However, we found that most of the influenza-virus-specific mAbs showed the highest affinity for the current vaccine strain. Thus, OAS does not seem to be a common occurrence in normal, healthy adults receiving influenza vaccination.     

恒河猴犬瘟热病毒基因疫苗制备单克隆抗体的研究

从感染犬瘟热病毒死亡的恒河猴肝脏组织中提取总RNA,经RT-PCR扩增H全基因并克隆到真核表达质粒pVAX中,构建了基因疫苗pVAX-H。用制备的基因疫苗免疫BaLb/c小鼠,小鼠可以产生免疫应答,免疫鼠脾细胞与SP2/0骨髓瘤细胞融合后,经间接ELISA法筛选,获得5株稳定分泌抗CDV H蛋白的单克隆抗体的杂交瘤细胞株。该5株单抗对应不同的病毒抗原表位,且均不与CPV和MV发生反应, McAb诱生的腹水可体外中和犬瘟热病毒,其中2株中和效价大于1:256。结果表明,CDV H基因疫苗可制备具有一定中和效价的抗CDV的单克隆抗体。     

五种赤潮藻单克隆抗体的制备

目的：建立分泌抗海洋褐胞藻(Chattonella marina)、卵圆褐胞藻(Chattonella ovata)、赤潮异弯藻(Heterosigma akashiwo)、具齿原甲藻(Prorocentrum dentatum)、尖刺拟菱形藻(Pseudonitzschia pungens)等5种藻的高特异性单克隆抗体杂交瘤细胞株．方法：利用细胞融合技术制备杂交瘤细胞,并通过反相间接血凝、间接酶联免疫吸附测定(ELISA)等方法筛选克隆阳性杂交瘤细胞株．结果：获得15CA7、23BG2和23BG9等3株杂交瘤细胞分泌的单克隆抗体与5种藻具有一定的特异性,但抗体的亲和力较低,仅与海洋褐胞藻具有较高的亲和力;而获得的M18杂交瘤细胞株分泌的单克隆抗体可与以上除尖刺拟菱形藻外4种藻产生高效价的明显交叉反应。结论：获得了3株分泌对5种藻具有一定的特异性较低亲和力抗体;而M18细胞株分泌的单抗与除尖刺拟菱形藻外4种藻产生明显的高效价的交叉反应,显示此单抗可代替5种海洋藻的抗血清．     

Demonstration by monoclonal antibodies that carbohydrate structures of glycoproteins and glycolipids are onco-developmental antigens

The hope that hybridoma antibodies would reveal unique cell surface antigens during embryogenesis, differentiation and oncogenesis has been replaced by the realization that such antigens are mainly carbohydrate structures of glycoproteins and glycolipids occurring in many cell types. These findings either may reflect limitations in the methods of selection of hybridoma antibodies or may point to important roles for the diverse carbohydrate structures as receptors for regulators of cell growth and differentiation.     

Monoclonal antibodies against carbohydrate differentiation antigens identify subsets of primary sensory neurones

Dorsal root ganglion (DRG) neurones transmit cutaneous sensory information from the periphery to the spinal cord. Within the dorsal horn of the spinal cord, classes of sensory fibres that are activated by different cutaneous stimuli terminate in separate and highly restricted laminae. Although the developmental events resulting in the laminar organization of sensory afferent terminals have not been defined, it is likely that interactions between surface molecules on DRG and dorsal horn neurones are involved in the generation of afferent synaptic connections. The identification of surface antigens that distinguish functional subclasses of DRG neurones would represent a first step in establishing the existence and nature of such molecules. We report here that monoclonal antibodies directed against carbohydrate differentiation antigens identify cytoplasmic and cell surface molecules expressed selectively by functional subsets of DRG neurons.     

结核分枝杆菌Hsp16.3单克隆抗体的制备及其特性鉴定

制备抗结核分枝杆菌Hsp16.3的单克隆抗体,并对其生物学特性进行鉴定。将含目的基因的表达载体pProEXHTb-Hsp16.3,通过E.coli DH5α诱导表达,获得含有6譎is的Hsp16.3蛋白,采用Ni-NTA纯化试剂盒进行目的蛋白的纯化,并用透析方法进行蛋白复性。将复性的蛋白免疫BALB/c小鼠,利用杂交瘤技术进行细胞融合,间接ELISA筛选阳性杂交瘤细胞株。将获得的Hsp16.3阳性杂交瘤细胞株分别利用间接ELISA法和Western blot方法进行效价、相对亲和力及特异性的测定。获得了三株Hsp16.3的单克隆抗体,分别命名为3H6F9、1D5E1和4H8G6,其效价分别为1:1?07、1:1?06和1:1?06,相对亲和力分别为0.0001 mg/mL、0.001 mg/mL和0.001 mg/mL,并且无交叉反应性。所获得的结核分枝杆菌Hsp16.3的单克隆抗体效价高、特异性强,为进一步研究Hsp16.3在结核分枝杆菌潜伏感染中的作用提供了有效的工具。     

抗棉铃虫组织蛋白酶B单克隆抗体的制备及鉴定

分别以棉铃虫组织表达的组织蛋白酶B和基因重组的大肠杆菌表达的组织蛋白酶B为抗原免疫BALB／c小鼠，选择血清抗体滴度高的免疫小鼠，取其脾细胞和Sp2／0骨髓瘤细胞进行细胞融合．经ELISA法筛选阳性克隆和有限稀释法克隆细胞，共获得分泌抗组织蛋白酶B的单克隆抗体杂交瘤细胞3株，分别命名为9D5，100E2和100H6．以ELISA法和Western blotting法对3株杂交瘤细胞株分泌的单克隆抗体进行了特异性鉴定，其中9D5和100E2能够同时识别两种不同来源的组织蛋白酶B，而100H6只能识别基因重组的大肠杆菌表达的组织蛋白酶B．     

Two contrary modes of chemolithotrophy in the same archaebacterium

Sulphur-dependent archaebacteria, which are found around nearly boiling continental solfataric springs and mud holes, can be assigned to two distinct branches: the aerobic, sulphur-oxidizing Sulfolobales and the strictly anaerobic sulphur-reducing Thermoproteales. Here, we report the isolation of a group of extremely thermophilic solfataric archaebacteria that are able to grow either strictly anaerobically by reduction, or fully aerobically by oxidation of molecular sulphur, depending on the oxygen supply. We have also established that the ability to grow in these two ways is shared by Sulfolobus brierleyi, a well-known less thermophilic sulphur-oxidizing archaebacterium capable of ore-leaching. The phenomenon may be dependent on a fundamental switch in genome expression. These organisms might represent the primitive fore-runners of sulphur-oxidizing archaebacteria, meeting their energy requirements either by oxidation or by reduction of the same element.     

Preferential utilization of the most JH-proximal VH gene segments in pre-B-cell lines

The most JH-proximal VH gene segments are used highly preferentially to form VHDJH rearrangements in pre-B-cell lines. This result demonstrates that the rate at which immunoglobulin VH gene segments recombine is influenced by their chromosomal organization, and that the initial repertoire of VH genes expressed in pre-B cells is strikingly different from that seen in mature populations.     

Two different gene loci related to the spatial patterning of brain ventricle in vertebrate

Observations on living embryonic brains and the microstructure of brain ventricle of goldfish revealed that there are two brain ventricle phenotypes in gynogenetic haploid embryos. One phenotype is as normal as that of the control inbreeding diploid embryos, which has normal differentiated forebrain, midbrain and hindbrain. Another phenotype is obviously abnormal, the brain patterning is irregular, and no distinct brain ventricle can be observed. The ratio of haploid embryos with normal brain pattern to that with abnormal brain pattern is 1:3. This ratio indicates that there are two gene loci involved in the spatial patterning of the brain ventricle. Since the possibility that deleterious recessive mutant alleles exist on both of the two gene loci had been excluded in this experiment, the phenotype represented the expressional state rather than the genotype of these two genes. Therefore, the ratio of 1:3 suggests that the expressing probability for each copy of the two genes is 50%, and the regulatory mechanism of the expression is based on two sets of chromosomes, controlled by the rule of the diploid-dependent regulatory mechanism.     

Two different gene loci related to the spatial patterning of brain ventricle in vertebrate

Observations on living embryonic brains and the microstructure of brain ventricle of goldfish revealed that there are two brain ventricle phenotypes in gynogenetic haploid embryos. One phenotype is as normal as that of the control inbreeding diploid embryos, which has normal differentiated forebrain, midbrain and hindbrain. Another phenotype is obviously abnormal, the brain patterning is irregular, and no distinct brain ventricle can be observed. The ratio of haploid embryos with normal brain pattern to that with abnormal brain pattern is 1∶3. This ratio indicates that there are two gene loci involved in the spatial patterning of the brain ventricle. Since the possibility that deleterious recessive mutant alleles exist on both of the two gene loci had been excluded in this experiment, the phenotype represented the expressional state rather than the genotype of these two genes. Therefore, the ratio of 1∶3 suggests that the expressing probability for each copy of the two genes is 50%, and the regulatory mechanism of the expression is based on two sets of chromosomes, controlled by the rule of the diploid-dependent regulatory mechanism.     

潍坊方言中的"同指异称"与"同称异指"现象

本文所论是潍坊方言中主要人际称谓的“同指异称”与“同称异指”现象。一方面,潍坊方言中祖父母、父母等主要称谓对象,各县市区有着不同的称谓用词;另一方面,潍坊方言中“爷爷”、“爷”等同一个称谓词,各县市区往往又代表着不同的称谓对象。本文就潍坊方言中人际称谓存在的这两方面的问题,作了较为全面的分析归纳。