乙脑E蛋白主要抗原片段与结核杆菌hsp70在毕赤酵母中的融合表达

目的构建乙脑E蛋白主要抗原片段与结核杆菌热休克蛋白70（Mycobacterium tuberculosis heat shock protein70，Mt．hsp70）基因融合表达载体，并研究其在巴斯德毕赤酵母中的表达情况，为改善发展新的乙脑疫苗打基础。方法以酵母表达载体pPICZa-A为基本单位构建E蛋白基因主要抗原片段与hsp70的融合表达载体，融合基因以酶切位点BamHⅠ连接，用电穿孔法转化酵母X-33，用Zeocin平板筛选重组子，经甲醇诱导表达后，SDS-PAGE和免疫印迹分析表达产物。结果E蛋白基因与hsp70的融合表达载体构建成功，SDS-PAGE显示，其相对分子质量为114kD，与实际大小相符，经凝胶薄层扫描分析，表达量为33mg／L，经Western印迹验证，有良好的抗原性。结论E—hsp70融合蛋白在酵母中的表达为下一步研究hsp70是否能作为E蛋白免疫时的理想佐剂作准备。     

Presence of two distinct acinar cell populations in human pancreas based on their antigenicity

Summary The immunohistochemical localization of ABH- and Lewis (Le)-related blood group antigens, including CA 19-9, a sialylated Le^a antigen, was examined using monoclonal antibodies (MoAbs) in 18 normal human pancreases and compared with ABH blood group antigenicity of the individuals. Acinar cells expressed ABH, Le^b, Le^y, and in some cases, Le^x antigen in various proportions, but not Le^a and CA 19-9. The reactivity of Le^b and Le^y was similar with regard to cellular localization and specificity. In all specimens but one, the distribution of Le^b (and Le^y) and H antigens on the one hand, and of A or B antigens on the other, showed a reciprocal relationship, in that one group of acini expressed Le^b (and Le^y) and H antigens, but lacked and A or B antigens (type 1 acinar cell); another group of acinar cells had A or B antigens, but expressed neither Le^b (Le^y) or H antigens (type 2 acinar cell). In ductal cells, four of eight individuals with blood group A, two of three with blood group B, and five of six with blood group O expressed the appropriate antigen, while the remainder did not. Le^a antigen was expressed primarily by centroacinar and terminal ductular and ductal cells of medium-sized ducts of all specimens, whereas Le^b was present in the cells of small and large ducts in all but four cases. The reactivity of ductal and ductular cells to Le^x was negative, except for one case. MoAb-Le^y and MoAb 19-9 reacted only with a few ductal cells in six (33%) and 12 cases (67%), respectively. There was no relationship in the expression of Le-related antigens between acinar and ductal/ductular cells; nor were there any sex difference with regard to the binding patterns of any antibodies. However, age appeared to influence the reactivity of some antibodies with acinar cells. Islet cells did not react with any of the antibodies. The results indicate that, although the antigenicity of epithelial cells can be affected by the host blood group types, there might be several regulatory systems for expression of blood group antigens in a cell-specific pattern.     

河北省麻疹野病毒抗原性变异分析

目的了解河北省麻疹野病毒与疫苗株抗原性之间的差异,为制定加速控制和消除麻疹的策略和措施提供科学依据。方法用麻疹疫苗免疫后的初免人群血清与河北省分离的麻疹野病毒（Hebei07-4和Hebei08-1）和麻疹疫苗株沪191分别进行中和试验,分析中和抗体之间的差异。结果中和抗体的均数为Hebei07-4 1∶16.77、Hebei08-1 1∶17.51、沪191 1∶157.15,初免人群血清中和麻疹野病毒的能力明显低于疫苗株。结论河北省麻疹野病毒与麻疹疫苗株的抗原性之间存在差异。     

肿瘤免疫学在未来面临的挑战

目前，人类肿瘤免疫学已进入了一个全新的的阶段，开始飞速发展。然而，在未来，肿瘤免疫学仍面临着许多挑战，仍有许多问题需要解决，例如人类肿瘤的抗原性问题，尚未阐明的机体对肿瘤免疫应答的细胞分子机制和免疫监视理论等等。     

Familial hereditary thrombocytopenia and HLA

Three generations of a Jewish family with hereditary thrombocytopenia (HT) are described. The disease was manifested clinically by mild bleeding tendency since infancy. Circumcision, however, did not result in excessive bleeding. HLA study in this family indicated that the HT locus is not linked to HLA.     

结核分支杆菌14KDa蛋白的基因克隆、表达、纯化及其抗原性的研究

目的基因克隆、表达、纯化结核分枝杆菌14KDa蛋白,研究其抗原性,评价其在血清学诊断中的价值。方法以结核分枝杆菌H37Rv基因组DNA为模板,应用PCR技术扩增14KDa蛋白基因片断,将其插入到高效表达载体PET-22b（＋）上,构建重组质粒PET-22b（＋）/14KDa,重组质粒在大肠杆菌中由IPTG诱导表达,表达产物经金属离子鳌合亲和层析方法纯化,免疫印迹和酶联免疫吸附（ELISA）试验分析重组蛋白的抗原性。结果构建了具有正确基因序列的14KDa蛋白重组质粒,在大肠杆菌BL21（DE3）中以可溶性蛋白形式表达,重组蛋白的表达量占菌体蛋白的40.8%,经过一步金属离子鳌合亲和层析后得到纯度为94.3%的目的蛋白。免疫印迹试验结果表明该蛋白能与羊抗结核血清发生特异免疫结合反应。应用ELISA方法对结核血清参考品进行检测,敏感性和特异性分别为75.6%和96.0%。结论获得了能高效表达14KDa蛋白的大肠杆菌工程菌,目的蛋白以可溶性形式表达,该重组蛋白具有良好的免疫原性,可望成为结核血清学的诊断抗原之一。     

Secretory granules of pituitary adenomas: quantitative study of hormonal antigenicity

Ultrastructurally, the antigenicity of major pituitary hormones in secretory granules was quantitatively investigated in five growth hormone (GH)-secreting adenomas, five prolactin (PRL)-secreting adenomas and eight clinically non-functioning (CN-F) adenomas. Sparsely granulated cells with a few or several small secretory granules (60–100 nm) exhibiting little or only weak antigenicity of various biochemically unrelated hormones were commonly observed in CN-F adenomas and occasionally in GH- and PRL-secreting adenomas. GH- or PRL-secreting adenomas consisted of many densely granulated cells with medium-sized (200–250 nm) or large (over 250 nm) secretory granules and a few or several sparsely granulated cells with small secretory granules. The densely granulated cells showed intense GH or PRL antigenicity and slight to moderate antigenicity for other hormones in large secretory granules and little or only weak antigenicity for various hormones including GH or PRL in small secretory granules. Their secretory granules larger than 160 nm or 140 nm significantly exhibited intense GH or PRL antigenicity (Fisher’s exact test; P < 0.05 and < 0.01, respectively). Two CN-F adenomas showed sparsely and densely granulated cells as well as intermediate cells. The densely granulated cells closely resembled GH-secreting cells. The intermediate cells simultaneously included small and medium-sized or large secretory granules exhibiting little/slight and intense GH-antigenicity, respectively. This study indicates that sparsely granulated cells of different categories showing slight antigenicity for various hormones, antigenically share the same origin, and that their hormonality, single or multiple, may be selectively activated in the developmental course of secretory granules. Received: 21 January 1999 / Revised, accepted: 13 July 1999     

浙江省近年来流感病毒株的抗原性及人群免疫状况分析

目的 为了解1998－1999年浙江省分离的流感病毒株的抗原性以及杭州市区人群对各类流感病毒的体液免疫力。方法 我们采用常规大量半加敏双向血凝抑制（HI）试验检测相应毒株的交叉HI滴度并计算相应的抗原比。用微量半加敏HI试验检测人群流感HI抗体滴度。结果 浙江省近2年来共分离到12株流感病毒代表株，其中11株为甲3（H3N2）亚型，1株为乙型。A3／浙防／10／98毒株与A3／汉防／359／95、A3悉尼／5／97、A3／沪防／1／98毒株的抗原比分别为1．4、4．0和16．0；A3／浙防／7／99毒株与上述3种标准毒株的抗原比分别为22．6、2．0和1．4。B／浙防／31／99毒株与B／京防／172／97和B／深防／12／97毒株的单向HI滴度分别为≥10240和40。杭州市区人群各型流感HI抗体阳性率为62．4％－100．0％，GMT在11．67－275．49之间。提示，除A3／浙防／10／98毒株抗原性类似于A3／汉防/359/95外，其余甲3亚型毒株的抗原性均类似于A3／沪防／1／98病毒。B型毒株抗原性属于巴拿马类病毒。结论 杭州市区人群对甲3亚型和B型巴拿马类毒株普遍具有HI抗体免疫力。     

乙型脑炎病毒SA14-14-2株NS1蛋白基因在原核细胞中的高效表达及其表达产物的抗原性分析

目的将乙型脑炎病毒NS1蛋白基因克隆至pET28-a( )表达载体,构建原核表达载体pET-NS1,并使该基因在E.coli中高效表达,为进一步研制乙脑早期诊断试剂奠定基础。方法根据GenBank中提供的乙型脑炎病毒SA14-14-2株全基因序列设计引物,通过反转录及巢式PCR方法扩增目的片段,测序后连接pET28-a( )载体,构建重组表达载体pET-NS1,转化大肠杆菌BL21(DE3)后,利用IPTG诱导获得高效表达。结果扩增出了1300bp的基因片段,与预期大小一致,测序后经Blast验证与已发表乙型脑炎病毒SA14-14-2株NS1蛋白基因序列同源性为100%,成功克隆至表达载体pET28-a( )并获得高效表达,表达产物分子量约为45kD,Western blot分析表明该表达产物具有良好抗原性。结论该表达产物的稳定高效表达及其抗原特异性为乙脑的诊断试剂开发提供了依据。     

松黄蛋白组分的分离及抗原性的实验分析

目的：分析松黄蛋白抗原中致敏成分。方法：用硫酸铵沉淀、葡聚糖凝胶层析分离松黄蛋白质组分，以致敏大鼠腹腔肥大细胞组胺释放试验和被动皮肤过敏反应试验检测松黄蛋白组分的致敏活性。结果：松黄蛋白中含有３种抗原性不同的组分，组分Ⅱ是松黄主要抗原。结论：松黄蛋白中含有致敏组分，此蛋白组分具对热和酸碱不稳定的性质。