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31.
Institute of Biochemistry, Academy of Sciences of the Uzbek SSR, Tashkent. (Presented by Academician of the Academy of Medical Sciences of the USSR Yu. A. Pankov.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 113, No. 2, pp. 168–170, February, 1992. 相似文献
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目的:制备抗醛糖还原酶(AR)的单克隆抗体(mAb),并与本室制备的抗醛糖还原酶相似蛋白(ARL-1)mAb进行比较。方法:经RT-PCR获得AR基因,将基因插入pGEX-4T-1(His)6C中,构建重组质粒pGEX-4T-1(His)6C-AR,以重组质粒转化E.coliRosetta诱导表达GST-AR蛋白。以纯化的GST-AR蛋白免疫BALB/c小鼠,采用杂交瘤技术制备mAb。应用间接ELISA和Western blot方法对mAb进行筛选和鉴定。使用Clustalx和Antheprot软件,比较AR与ARL-1的同源性,表达GST-dAR[80~142氨基酸(aa)],与ARL-1差异较大;并分析AR的抗原性,表达GST-dA1(1~79aa)、GST-dA2(80~99aa)、GST-dA3(111~142aa)、GST-dA4(143~316aa)。利用AR全长及截短蛋白,采用Western blot分析制备的抗AR mAb识别AR抗原的部位。结果:获得3株稳定分泌抗AR mAb的杂交瘤细胞系ARB3、AR7B3G4和ARF10。3株抗GST-AR的mAb均为IgG1(κ型),腹水mAb效价为1∶4×105,细胞培养上清mAb效价为1∶1×104,3株mAb均可与胎盘组织中的AR蛋白起反应,而与GST-ARL-1和GST蛋白无交叉反应。它们分别为抗GST-dA1、GST-dA3和GST-dA4蛋白的mAb。结论:成功地制备了3株特异性抗AR mAb,可分别识别AR的1~79、111~142、143~316位氨基酸。将它们与抗ARL-1mAb联合应用,将有助于进一步研究AR与ARL-1蛋白的功能,并为深入探讨AR、ARL-1与相关疾病的关系及进行大规模的流行病学调查提供了有力的工具。 相似文献
34.
人膀胱癌T24细胞及其SCID小鼠移植瘤组织中KDR的表达 总被引:1,自引:1,他引:0
目的 检测KDR在人膀胱癌T24细胞及其SCID小鼠移植瘤组织中的表达。方法 常规培养T24细胞,建立荷人膀胱癌移植瘤SCID小鼠动物模型,使用抗KDR单克隆抗体,通过免疫荧光、免疫组化检测KDR在T24细胞及瘤体组织上的表达;结果 KDR在T24细胞及瘤体组织中均呈阳性表达。结论 本研究结果为进一步研究抗KDR单抗在荷T24细胞移植瘤动物体内分布及对瘤体生长的抑制效应奠定了实验基础。 相似文献
35.
WHAN KOOK CHUNG KYU YONG CHOI CHANG DON LEE JIN WU CHUNG HEE SIK SUN KYU WON CHUNG BOO SUNG KIM CHUNG SIK CHUN KYOO HONG CHO SEUNG JO KIM 《Journal of gastroenterology and hepatology》1987,2(1):13-17
Different doses of hepatitis B virus vaccine—prepared by Korea Green Cross Corporation, were given to healthy infants born to HBsAg-negative mothers at birth, 1 and 6 months of age. A dose of 2 μg was administered intradermally in Group A and, in the three other groups, the vaccine was given intramuscularly (i.m.). An adequate follow-up observation was possible for 9 months after birth in 22, 25, 23 and 21 infants in Groups A, B, C and D, respecvely.
Group C (5 μg, i.m.) produced seroconversion most rapidly, showing the highest rate (96%) at 9 months of age. The lowest seroconversion rate (5%) was found at the age of 1 month in Group A subjects, but the rate increased to 91% after a booster dose was given at 6 months of age.
While it can be concluded that a 5 μg i.m. dose of vaccine at 0, 1 and 6 months of age is optimum for the immunization of infants in efficacy and economy, a 2 μg intradermal dose can also be considered as an immunogenic and economical regimen, though the immune response is slower and a special technique is required for immunization. 相似文献
Group C (5 μg, i.m.) produced seroconversion most rapidly, showing the highest rate (96%) at 9 months of age. The lowest seroconversion rate (5%) was found at the age of 1 month in Group A subjects, but the rate increased to 91% after a booster dose was given at 6 months of age.
While it can be concluded that a 5 μg i.m. dose of vaccine at 0, 1 and 6 months of age is optimum for the immunization of infants in efficacy and economy, a 2 μg intradermal dose can also be considered as an immunogenic and economical regimen, though the immune response is slower and a special technique is required for immunization. 相似文献
36.
Isolation and characterization of a monoclonal anti-P30 antibody resistant mutant of Toxoplasma gondii 总被引:13,自引:0,他引:13
LLOYD H. KASPER 《Parasite immunology》1987,9(4):433-445
Of the possible iodine-labelled Toxoplasma gondii surface proteins, P30 (apparent Mr 30,000) is the principal one recognized by acute and convalescent anti-toxoplasma sera. This protein which comprises from 3 to 5% of the total parasite protein was used to raise a panel of parasiticidal monoclonal anti-P30 antibodies. One of these monoclonal antibodies was able to select a resistant mutant from a large population of chemically mutagenized wild-type P strain parasites. This mutant retained the wild type sensitivity to other non-P30 parasiticidal monoclonal antibodies as well as polyclonal anti-P30 rabbit sera. Analysis of surface radioiodinated wild type and mutant parasites showed that the mutant had a quantitative reduction in the amount of P30. A comparison of surface biotin labelled wild type and resistant parasites by two dimensional electrophoresis showed that the mutant lacked one and possibly two of several proteins that make up wild type P30. Western blot analysis indicated that the mutant was devoid of antigenically reactive P30. These findings further support the hypothesis that antigenic variants of T. gondii can be induced and may involve the major surface membrane antigens of the parasite. 相似文献
37.
A monoclonal antibody against choline acetyltransferase (ChAT), the acetylcholine-synthesizing enzyme, was used to study cholinergic synapses on identified (Golgi stained) granule cells in the rat fascia dentata. Choline acetyltransferase immunocytochemistry was applied to 40-microns Vibratome sections cut perpendicular to the longitudinal axis of the hippocampus. Light microscopy revealed fine varicose ChAT-immunoreactive axons in all layers of the fascia dentata, i.e., in the stratum moleculare, the stratum granulosum, and the subgranular polymorph zone. Most fibers were observed in the vicinity of granule cell bodies where they ran mainly parallel to the granular layer. Next, the immunostained Vibratome sections were sandwiched between small pieces of Parafilm and piled to form a block that was covered with agar and Golgi stained. After that, the sections were separated by cutting away the agar and removing the Parafilm. Sections containing well-impregnated granule cells were gold-toned (Fairén et al., '77), embedded in Araldite, and subjected to ultrathin sectioning for electron microscopy. A total of 14 gold-toned granule cells were examined in the electron microscope for synaptic contacts with cholinergic afferents. Choline acetyltransferase-immunoreactive axon terminals were observed that established symmetric synaptic contacts with the cell bodies and dendritic shafts of the gold-toned identified granule cells. Two types of contact were observed on spines arising from gold-toned granule cell dendrites. Immunoreactive terminals established asymmetric synaptic contacts with the head of small spines and symmetric contacts with the stalk of large, complex spines. The boutons forming asymmetric synaptic contacts with the cup-shaped spine head of the complex spines were not found to be immunoreactive. Our results demonstrate that cholinergic fibers to the rat fascia dentata establish characteristic types of synaptic contact with different postsynaptic elements of granule cells, suggesting a complex function of this afferent system. 相似文献
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免疫吸附治疗在肾移植致敏受者中的应用 总被引:2,自引:0,他引:2
目的:探讨蛋白A免疫吸附(immunoads Orption,IA)治疗对清除肾移植致敏受者体内特异性抗HLA抗体的疗效和安全性。方法:10例肾移植致敏(PRA〉50%)受者用彤新蛋白A免疫吸附柱行IA治疗,测定治疗前后血免疫球蛋白及PRA水平。结果:10例患者IA治疗次数为4~15次(中位数9)。所有患者IA治疗后血清总IgG水平都明显下降(P〈0.001),IgA和IgM也较治疗前显著降低(P〈0.01)。PRA8例转阴,1例〈30%,1例仍为100%。结论:对于肾移植致敏受者,IA是一种特异性高、安全有效的治疗措施。 相似文献
40.