抗人结肠癌相关抗原单克隆抗体的制备及初步研究

目的：制备抗人结肠癌相关抗原的单克隆抗体4D10，运用组织芯片技术研究其与人结肠癌组织反应的情况。方汪：用人结肠癌细胞株LOVO免疫小鼠，用间接细胞ELISA和免疫组化的方法筛选抗结肠癌相关抗原的单抗。结果：获得一株能够稳定分泌抗人结肠癌相关抗原的杂交瘤细胞株。4D10能够与不同分期的结肠癌组织切片反应，且与正常组织和其他癌组织几乎不反应。结论：4D10对结肠癌的病理分期及预后结果都具有一定的临床参考价值。     

Immunohistological characterization of a monoclonal antibody (OV632) against epithelial ovarian carcinomas

Summary A hybridoma cell line (OV632) producing monoclonal antibody against ovarian carcinomas was developed from the spleen cells of a mouse immunized with cystic fluid from a serous cystadenocarcinoma. Immunohistological studies in frozen sections showed that 22 out of 28 nonmucinous ovarian carcinomas, which included serous, endometrioid, clear cell, and undifferentiated tumours, reacted with this antibody. Three out of 7 mucinous ovarian carcinomas were positive, whereas only 7 out of 122 extra-genital malignant lesions, predominantly adenocarcinomas, were positive. The negative cases included 38 breast carcinomas and 24 colon carcinomas, tumours which are responsible for most of metastatic disease in the ovary. On the basis of these findings, the antibody OV632 is considered appropriate for histodiagnostic purposes as an aid in the distinction between primary and secondary ovarian cancer.     

戊型肝炎病毒第Ⅳ基因型毒株中和抗原表位的鉴定

目的确定在我国新发现的戊型肝炎病毒（HEV）第Ⅳ基因型毒株的中和抗原表位特征及其与世界各地其他基因型HEV毒株中和抗原表位的异同。方法制备HEV第Ⅳ基因型ORF2重组衣壳蛋白p166Chn及其单克隆抗体（McAb），采用体外中和试验鉴定McAb的中和活性；通过间接ELISA和免疫印迹法测定中和性McAb与不同基因型HEVORF2编码蛋白p166的免疫反应性，并结合相加ELISA确定p166Chn中和抗原表位的分布和性质。结果获得6株稳定分泌抗-p166Chn McAb的杂交瘤细胞株。所得McAb能在体外中和HEV中国毒株（第Ⅳ基因型）对PLC/PRF／5细胞的感染性，并与来源于HEV4种不同基因型代表株的7种p166重组蛋白（p166Chn、p166Bur、p166Mor、p166Pak、p166Mex、p166Us和p166Nz）均能发生阳性反应。抗体间的相加ELISA结果为阴性。表明6株McAb识别p166上的同一个中和抗原表位。结论在我国新发现的HEV第Ⅳ基因型毒株与分布在世界各地的其他基因型毒株具有共同的中和抗原表位。     

An ELISA that detects cells-associated and released rat IL-2 receptors in a soluble form

A mouse anti-rat interleukin-2 receptor (IL-2R) monoclonal antibody (mAb), ART-65, has recently been developed which recognizes the rat IL-2R but binds to an epitope distinct from that recognized by the mAb ART-18. The availability of two mAb against the rat IL-2R molecule permitted the development of an enzyme-linked immunosorbent assay (ELISA) which detects soluble and solubilized rat IL-2R. Using this ELISA, time course studies of ConA and LPS-activated rat splenocytes revealed that IL-2R are released into their environment during the culture period, and that IL-2R can be detected in rat sera. The significance of the results is discussed.     

用间接ELISA检测抗H9亚型AIV独特型单克隆抗体的研究

目的为制备抗H9亚型A型流感病毒(AIV)独特型杂交瘤细胞系,运用异种动物间共有独特型理论建立的间接ELISA进行检测.方法本试验采用兔抗H9亚型低致病力AIV IgG作为检测抗原,通过预试验确定了包被抗原的工作浓度为800μg/mL,以间接ELISA方法检测融合细胞培养上清液,筛选到产生抗AIV鸡和兔种间共有独特型抗体的杂交瘤细胞.结果间接ELISA试验中P/N值达12,效价达到2-4,证明杂交瘤细胞株分泌目的单克隆抗体的能力强.结论该检测方法排除了分泌抗鸡同种型表位以及超变区其他表位抗体的杂交瘤细胞,从而成为筛选分泌抗H9亚型AIV独特型单克隆抗体杂交瘤细胞株的可靠方法.     

Monoclonal antibodies against CEA-related components discriminate between pancreatic duct type carcinomas and nonneoplastic duct lesions as well as nonduct type neoplasias

Summary The expression of CEA and related antigens in formalin-fixed paraffin-embedded tissues of normal pancreas and different pancreatic neoplasms was studied immunocytochemically using three monoclonal antibodies (MAbs) recognizing different epitopes on CEA and related antigens. Additionally, a number of extrapancreatic malignancies were tested. The epitope recognized by MAb 250 (present on CEA and NCA 95) was expressed in all but one pancreatic ductal adenocarcinoma and ampullary carcinoma (42/43). The MAb 431 defined epitope (present only on CEA) was less frequently found (27/43). MAb 374, defining an epitope on CEA, NCA 95 and NCA 55 proved to be nearly as sensitive tive as MAb 250, but also reacted with normal duct epithelium. In contrast, MAb 250 and MAb 431 discriminated clearly between reactive duct lesions and malignant duct changes. Moreover, these MAbs differentiated between pancreatic duct carcinomas and nonduct type carcinomas as well as benign pancreatic tumours. In duct type carcinomas, the strongest staining was observed in well differentiated tumours. No discrimination was possible between pancreatic carcinomas and other adenocarcinomas of the gastrointestinal tract nor between most of the lung carcinomas and some other malignancies, specified below. MAb 250 and MAb 431 failed to react with hepatocellular carcinomas, renal cell carcinomas, carcinoids, sarcomas and melanomas. The findings suggest that paraffin-embedded tissues of pancreatic duct type carcinomas, in contrast to nonduct type tumours and normal ducts, are distinguished by the presence of a CEA and NCA 95 related epitope.Presented in part at the American Pancreatic Association Meeting, Chicago, 1984Supported by Deutsche Forschungsgemeinschaft Kl 366/4-1 and SFB 215     

Egf/r3单克隆抗体治疗肺腺癌的实验研究

目的非小细胞肺癌(NSCLC)存在表皮生长因子受体(EGFR)过度表达。Egf/r3为EGFRMcAb,属IgC2a亚型,具有一定的治疗肺癌作用,但其机理尚不完全清楚。为更进一步全面了解Egf/r3对肺癌的治疗作用,本文以高表达EGFR的SPC-A-1和A549肺腺癌细胞系做为靶细胞,观察Egf/r3对肺腺癌细胞周期的影响、Egf/r3与CDDP和8-Mop是否存在协同抗肺癌作用。方法细胞周期检测采用流式细胞仪PI染色法分析,协同作用采用MTT法。结果SPC-A-1和A549G1期细胞分别增加5·0%和7·8%,S期分别减少5·6%和8·6%,G2-M期变化不明显。Egf/r3分别与CDDP和8-Mop联用,对SPC-A-1和A549杀伤率分别达到35%、39%和36%、39·5%。结论Egf/r3阻抑G1期癌细胞进入S期,使细胞同步化,增加了靶细胞对化疗药物的敏感性,减少癌细胞化疗后损伤修复,与CDDP、8-Mop存在协同抗肺腺癌作用。     

美蓝抗胃癌单克隆抗体在胃癌术中的淋巴结标记

目的　探讨胃癌术中淋巴结的标记方法。方法　 2 2 4例胃癌患者随机分为 3个组 :美蓝 抗胃癌单克隆抗体 (MAb3H 11)注射组 (观察组 ) 76例 ;美蓝组 73例 ;对照组 75例。结果　观察组 76例清扫淋巴结 2 2 0 1个 ,平均每例清扫 2 9.0个 ,平均每例清扫转移淋巴结 7.8个 ( 5 93 / 76) ;美蓝组 73例清扫淋巴结 1619个 ,平均每例清扫 2 2 .2个 ,平均每例清扫转移淋巴结 6.0个( 4 41/ 73 ) ;对照组 75例清扫淋巴结 13 77个 ,平均每例清扫 18.4个 ,平均每例清扫转移淋巴结 4.9个 ( 3 68/ 75 ) ;平均每例清扫的淋巴结数及转移淋巴结数 ,观察组均明显多于美蓝组和对照组 (P <0 .0 1)。结论　应用美蓝 抗胃癌单克隆抗体 (MAb 3H 11)术中胃壁注射 ,可有效的标记胃的区域淋巴结     

抗人钠碘转运体单克隆抗体杂交瘤细胞系的建立与初步鉴定

目的 制备抗人钠碘转运体(NIS)单克隆抗体。方法 以人NIS蛋白质的二个片段(第2膜外段和第14f段)为抗原免疫小鼠，运用杂交瘤技术，制备鼠抗人NIS单克隆抗体(rhNIS MAb)，采用小鼠Ig亚类测定试剂条测定单抗的亚类，并通过ELISA、Western-blot、免疫组化技术等鉴定其特异性。结果 运用杂交瘤技术成功地制备出能稳定分泌抗人NIS单克隆抗体的细胞系。Western-blot分析表明：此单抗所识别的靶分子的相对分子量主要在97KD，免疫组化染色显示：甲状腺滤泡细胞基底膜上有棕黄色着色。结论 成功地制备出抗人NIS单克隆抗体，为NIS抗原-抗体的研究提供了新的工具，NIS单克隆抗体不仅可以应用于甲状腺、乳腺等领域的基础研究，而且可以用于临床，为甲状腺疾病的诊断和治疗提供切实保障。     

Snake venom metalloproteinases: structure/function relationships studies using monoclonal antibodies.

Snake Venom Metalloproteinases (SVMPs) are synthesized as zymogens and undergo proteolytic processing resulting in a variety of multifunctional proteins. Jararhagin is a P-III SVMP, isolated from the venom of Bothrops jararaca, comprising metalloproteinase, disintegrin-like and cysteine-rich domains. The catalytic domain is responsible for the hemorrhagic activity. The disintegrin-like/cysteine-rich domains block alpha2beta1 integrin binding to collagen and apparently enhance the hemorrhagic activity of SVMPs. The relevance of disintegrin-like domain is described in this paper using a series of mouse anti-jararhagin monoclonal antibodies (MAJar 1-7). MAJar 3 was the only antibody able to completely neutralize jararhagin hemorrhagic activity. Neutralization of catalytic activity was partial by incubation with MAJar 1. MAJars 1 and 3 efficiently neutralized jararhagin binding to collagen with IC50 of 330 and 8.4 nM, respectively. MAJars 1 and 3 recognized the C-terminal portion of the disintegrin domain, which is apparently in conformational proximity with the catalytic domain according to additivity tests. These data suggest that disintegrin-like domain epitopes are in close contact with catalytic site or functionally modulate the expression of hemorrhagic activity in SVMPs.