Studies on monoclonal anti-isotypic and anti-idiotypic antibodies against leukemia and myeloma: V. The effects of monoclonal antibodies and interferon on the levels of cyclic nucleotides in leukemic cell lines

Summary After the leukemic cell lines were treated with monoclonal antibodies (McAbs) and interferon (IFN-α), the changes of cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP) levels in the corresponding leukemic cell lines were measured by radioimmunoassay. The results showed that when the ratio of antigen to antibody was 80 to 1, the cAMP levels in the leukemic cell lines were obviously higher than those in the controls while the cGMP levels were obviously lower after being treated with the corresponding McAbs for 16–24 h (P<0.001). The average level of intracellular cAMP was remarkably increased and that of cGMP underwent no significant changes in the leukemic cell lines after treatment with IFN-α. This work was supported by grant no. 38707803 from the National Natural Sciences Foundation of China and from the Young Teachers Foundation of National Educational Committee of China.     

一组抗人肝癌单抗相应抗原的表达及位点分析

为分析肿瘤细胞的异质性，解决导向诊治中的问题。方法：我们用流工细胞仪定量地测定了四株抗人肝癌单抗在五种不同人肝癌细胞株ＳＭＭＣ－７７２１，ＢＥＬ－７４０２，ＱＧＹ－７７０１人肝癌细胞系ＨＨＣＣ，ＨＨＣＣ－９２０４和正常人肝细胞ＱＺＧ中的相应抗原表达及抗原位点分析。结果：Ｈ１８，Ｈ２７和Ｅ５相应抗原在五种人肝癌细胞株中均有表达，Ｆ１１在ＳＭＭＣ－７７２１，ＱＧＹ－７７０１两种肝癌细胞株有表达，但表达     

Preparation and antigenic characterization of rat monoclonal antibodies againts Hantavirus

ＰｒｅｐａｒａｔｉｏｎａｎｄａｎｔｉｇｅｎｉｃｃｈａｒａｃｔｅｒｉｚａｔｉｏｎｏｆｒａｔｍｏｎｏｃｌｏｎａｌａｎｔｉｂｏｄｉｅｓａｇａｉｎｔｓＨａｎｔａｖｉｒｕｓ￥ＸｕＨａｉｆｅｎｇ（徐海峰）；ＬｉｕＹｕａｎ（刘原）；ＬｉａｎｇＺｈｅ（梁喆）；ＭｉＬ...     

人脑肌酸激酶单克隆抗体的制备及鉴定

用事故死亡者脑组织中提纯的人脑肌酸激酶（ＣＫ－ＢＢ）对ＢＡＬＢ／Ｃ小鼠进行免疫，应用杂交瘤技术将免疫小鼠脾细胞分别与２种骨髓瘤细胞（ＮＳ－１，６５３）融合，筛选得到８株能稳定分泌ＩｇＭ抗体的杂交瘤细胞。对分泌抗体经酶联免疫电泳转移试验（Ｗｅｓｔ－Ｂｌｏｔ）进行了鉴定，其中４种抗体能特异地与ＣＫ－ＢＢ的ＳＤＳ－ＰＡＧＥ电泳带结合。提示这些单克隆抗体有可能应用于肌酸激酶的结构功能研究和临床检验中。     

抗人IgM单克隆抗体的制备及其特征研究

以人IgM作为抗原免疫Balb/C小鼠,采用杂交瘤技术获得5株分泌抗人IgM单抗的杂交瘤细胞株.通过酶联免疫吸附试验、免疫双向扩散试验证明:该组单抗仅与人IgM起反应,而不与其他各类免疫球蛋白反应。间接免疫荧光试验证明:该组单抗测得不同人群外周血淋巴细胞阳性数与用抗B细胞单抗和直接免疫荧光法测得的结果接近.免疫转印试验证明抗体作用的分子量为50～70kd(dalten法定单位为u,换算系数0.9921,下同).提示为IgM.本文还对单抗应用价值进行了讨论。     

Studies on monoclonal anti-isotypic and anti-idiotypic antibodies against leukemia and myeloma: I. preparation and characteristics of monoclonal anti-isotypic antibodies to igm from B chronic lymphocytic leukemia

Five hybridomas producing McAbs against human serum IgM-isotype were obtained by fusing the myeloma NS-1 cell and the RALB/C murine spleen cell which had been immunized by serum IgM of patient with B chronic lymphocytic leukemia (B-CLL). These McAbs only reacted with human IgM, not with other immunoglobulins in ELISA and immune double diffusion test. An approximate positive rate of peripheral blood lymphocyte (PBL) was got when these McAbs and the McAbs aginst B cell were tested by indirecl immunofluorescent assay. The positive rate was similar to that obtained by direct immunofluorescent test. Immunoblotting showed that the molecular weight of the antigen to these McAbs was 70–90 Kd, indicating that the antigen was IgM. The practical value of the McAbs against human IgM was also discussed.     

微波对白血病耐药细胞株K562/MDR的影响

目的 研究微波对白血病耐药细胞株K562／MDR耐药性的影响。方法 使用流式细胞仪测量K562／MDR细胞内罗丹明（Rh123)含量及多药耐药基因产物P－糖蛋白（P－gp)的表达，使用DPH标记的荧光分光光度计法测量K562／MDR细胞膜流动性。结果 微波处理后，K562／MDR细胞内罗丹明含量增加，P－gp糖蛋白表达下降，细胞膜流动性增加。结论 微波能对白血病耐药细胞株K562／MDR的耐药性起一定的逆转作用。     

分泌抗心脏肌凝蛋白及其轻链单克隆抗体杂交瘤细胞系的建立及初步鉴定

心脏肌凝蛋白(Cardiac myosin,CM)及其轻链(CM—LC)是心肌特有的收缩蛋白质,其化学结构和免疫学性质均与其它组织来源者不同。当急性心肌梗塞(AMI)造成心肌细胞缺血坏死时,血清CM—LC水平升高。故可用于临床诊断AMI。这一诊断指标的特异性、敏感性、与梗塞灶大小     

Studies on monoclonal anti-idiotypic antibodies to human B cell leukemia

Murine immunocytoma cell line, NS-1, was fused with spleen cells of Balb/C mice which had been stimulated by tolerogenic disaggregated human gamma globulin and immunized by purified serum IgM from the patient with chronic B cell leukemia (B-CLL). 10 hybridoma cell lines secreting monoclonal anti-idiotype (anti-Id) antibodies to human CLL were obtained. The McAbs were subclasses belonging to IgM and of IgG mouse. Specificity and biologic characters of the monoclonal anti-Id antibodies from culture fluid or ascites were assayed by ELISA, indirect mixed ELISA sandwich, ELISA inhibition, immunofluorescence (IF) and IF inhibition. The study also proved that monoclonal anti-Id antibodies could react with homologous IgM, but not with Ig from normal donors or a panel of patients with myeloma. The results of IF and IF inhibition assay showed that monoclonal anti-Id antibodies were bound to lymphocytes of patient with B-CLL. Their reactivity was inhibited by homologous IgM, but not by lymphocytes of patients with ALL or lymphoma. Monoclonal anti-Id antibodies were heterogenous reactive patterns with cell lines in vitro.     

SjAWMMcAb对吡喹酮杀虫效果的影响

目的 :制备日本血吸虫成虫表膜单克隆抗体 (SjAWMMcAb) ,探讨其与吡喹酮在宿主体内的联合杀虫作用。方法 :用SP2 / 0骨髓瘤细胞与日本血吸虫成虫表膜抗原免疫的BALB/c小鼠脾细胞融合 ,经筛选和克隆化后建立分泌日本血吸虫成虫表膜抗原单克隆抗体杂交瘤细胞株 ,通过单层细胞培养法和小鼠体内接种法收集单克隆抗体 ;采用Westernblotting和免疫荧光测定 (IFA)对其进行鉴定 ;小鼠实验观察单抗与吡喹酮的联合杀虫作用。结果 :建立了 3个分泌SjAWMMcAb的细胞株 ,Westernblotting显示 3个McAb可识别 5种不同分子量的蛋白 ,IFA表明 3个McAb均能与血吸虫成虫表膜发生特异性反应 ,3B6和 1C2两株McAbs与吡喹酮联用杀虫率分别可提高 4 1.78%和 2 4 .12 %。结论 :某些SjAWMMcAb与吡喹酮可发挥联合杀虫作用     

分泌抗体的杂交瘤细胞株的制备

目的应用杂交瘤技术制备出分泌抗肺癌抗体杂交瘤细胞株,为肺癌的早期诊断和免疫治疗提供可能靶点．方法用肺癌细胞株作免疫原免疫健康BALB／C小鼠,取其脾细胞与小鼠骨髓瘤细胞融合,获得分泌高滴度抗肺癌抗体的杂交瘤细胞株,用间接ELISA和检测单抗体的特异性,双抗体夹心法测定免疫球蛋白同种型．结果通过细胞融合,筛选出2株持续分泌抗肺癌抗体的杂交瘤细胞株C5、B8,其分泌的抗体为IgM亚类,κ亚型,并和肺癌细胞发生特异性反应．结论成功完成细胞融合,制备了分泌抗体的杂交瘤细胞株．     

抗人精子单克隆抗体的制备和特性鉴定

用杂交瘤技术制备了12株稳定地分泌抗人精子单克隆抗体的杂交瘤细胞株,用间接免疫荧光法观察这些抗体分别结合到精子不同部位,有7个抗体使精子产生头—头凝集或尾—尾凝集,有4个抗体使精子制动,只有1个抗体与淋巴细胞有交叉反应。测定了10个单克隆抗体的免疫球蛋白亚类,7个属于IgG,13个属于IgM。     

细粒棘球绦虫囊液McAb的制备及鉴定

McAbs were prepared by fusing SP2/0 cells with spleen cells of BALB/c mice immunized with hydatid cyst fluid (HCF) antigen. Antibodies in hybrid culture supernatant were detected by ELISA, and positive hybrids were cloned by technique of limiting dilutions. Numbers of chromosomes of five hybridoma cell lines (1D1, 1D8, 2D6, 4B10, 4F4) were demonstrated in the range of 99-108. The immunoglobulin class/subclass of McAb 4F4 was IgM and the others belonged to IgG1. IHA, ELISA and I-ELISA were used for identifying titer and specificity of five McAbs. The titers of two McAbs (1D1, 1D8) reached 1:10(6)-10(7) in IHA and ELISA. Two of these McAbs (1D1, 4F4) with other parasitic antigens revealed cross-reactions. This indicated that the McAbs 1D1 and 4F4 were against common antigen determiners of different parasitic antigens and the McAbs 1D8, 2D6, 4B10 against specific antigen determiners of HCF.     

细粒棘球绦虫囊液McAb的制备及鉴定

作者用人源包虫囊液抗原免疫的BALB/c小鼠脾细胞与SP 2/0骨髓瘤细胞杂交,获得成功。融合率100％,抗体阳性率10.7％。经克隆化培养获得分泌抗人源细粒棘球绦虫抗原单克隆抗体的杂交瘤细胞5株,其染色体数为99～108。经鉴定其中3株(1D_3、2D_6、4B_(10))是特异针对细粒棘球绦虫抗原决定簇的,均属IgG_1亚类;另外2株(4F_4、1D_1)则针对细粒棘球绦虫与其他几种寄生虫抗原的共同决定簇,它们分属IgM和IgG_1类免疫球蛋白。     

B7H3分子在人髓性白血病细胞株U937和K562上的表达及生物学意义

目的:研究共刺激分子B7H3分子在人髓性白血病细胞株U937和K562上的表达及其生物学意义?方法:应用流式细胞术及RT-PCR法检测B7H3分子在人髓性白血病细胞株U937和K562上的表达;并用鼠抗人B7H3单克隆抗体(mAb)作用于U937和K562细胞株,细胞计数法检测鼠抗人mAb B7H3对U937和K562细胞株生长的影响?结果:流式细胞术分析显示U937和K562细胞株均表达B7H3膜蛋白,RT-PCR检测到U937和K562细胞有B7H3 mRNA产物;鼠抗人mAb B7H3对U937和K562细胞株有抑制生长的作用?结论:U937和K562细胞株组成性表达B7H3分子;鼠抗人mAb B7H3与U937和K562细胞表面的B7H3分子交联后可以抑制白血病细胞的生长?     

一组抗人髓系细胞分化抗原单克隆抗体的制备和初步应用

用新鲜白血病细胞或细胞株免疫BALB/c小鼠,经细胞融合技术制备得6株能稳定分泌抗人髓系细胞分化抗原单克隆抗体(以下简称单抗)的杂交瘤株.经鉴定该组单抗各自所识别的特异性抗原分子量为46～150kd.这些抗原在粒单系细胞膜上均有程度不等的表达,6种单抗中有5种具有结合兔补体的能力,其对白血病细胞株补体依赖的特异性细胞毒为73%～94%.两种单抗(Zh_(820),Zh_(114))对正常CFU-GM生长有抑制作用外,余4种单抗无明显影响.6种单抗均与FAB M_1～M_5型急性髓系白血病细胞有不同程度的阳性反应.     

肾综合征出血热病毒大鼠单克隆抗体制备及其对病毒抗原特性的鉴定

用肾综合征出血热病毒（ＨＦＲＳＶ）免疫Ｌｏｕ／ｃ大鼠脾细胞与ＩＲ９８３Ｆ骨髓瘤细胞融合，获得１１株能分泌特异性抗体的杂交瘤细胞系。用放射免疫沉淀、Ｗｅｓｔｅｒｎｂｌｏｔｔｉｎｇ及免疫酶技术等对这一组大鼠单克隆抗体（单抗）进行了特性鉴定和病毒抗原性分析。发现１１株单抗中，９株为抗核壳蛋白（ＮＰ）特异性，２株为抗Ｇ２糖蛋白。大多数抗ＮＰ单抗无中和活性及血凝抑制（ＨＩ）活性，但有２株例外，具有一定程度的中和或（和）ＨＩ活性。而抗Ｇ２单抗均只有一定程度的中和或（和）ＨＩ活性。在ＨＦＲＳＶ抗原性分析方面，可初步将１１株大鼠单抗分成组、亚组、野鼠型及亚型特异性单抗，并对上述单抗特性进行了讨论。     

Study and Application of Monoclonal Antibodies against Virus of Haemorrhagic Fever with Renal Syndrome

This paper summarizes the study and application of monoclonal antibodies (McAbs)against haemorrhagic fever with renal syndrome (HFRS) virus in our department over the past sever-al years. The following six points are discussed: (1) establishment and characterization of thehybridoma cell lines secreting McAbs against HFRS virus and hemagglutinin of the virus: (2) theantigenic analysis of HFRS viruses in China by McAbs: (3) purification and application of theMcAbs: (4) purification and characterization of HFRS virus 50K stnactural protein by McAba-affini-ty chromatography and the McAbs possessing different characteristics: (5) detection of HFRS virusantigen in peripheral blood lymphocytes from HFRS patients by the McAb-IFAT; and (6)development of McAb-ELISA indirect sandwich methods, and detection of HFRS virus antigen andIgM, IgG and/or HI antibodies in human and animals. The results of the studies show that theMcAbs can be used for early diagnosis, epidemiological investigation, preparation of vaccine andimmunotherapy of HFRS.     

抗赭曲霉毒素A单克隆抗体的制备及其ELISA检测方法的建立

目的制备抗赭曲霉毒素A（OA）的单克隆抗体（McAb）并对其进行初步鉴定,在此基础上建立竞争抑制酶联免疫吸附试验（ELISA）用于OA的检测。方法采用小剂量长周期的免疫方案,以OA 牛血清白蛋白（BSA）偶联物免疫雌性BALB/c小鼠,采用细胞融合法获得分泌抗OA的McAb的杂交瘤细胞株,用竞争抑制ELISA法进一步检测McAb的特异性,腹水诱生法大量制备McAb,以OA为竞争抗原,建立检测OA的竞争抑制ELISA。结果OA BSA免疫的BALB/c小鼠血清效价为1:512?000,与BSA有强烈的交叉反应。细胞融合后,ELISA筛选抗体分泌阳性的杂交瘤细胞株,抗OA BSA的McAb与BSA的交叉反应率仅为3.5%,对分泌抗OA BSA特异的McAb的细胞株经3轮克隆化,抗体分泌阳性率达到100%,建立了1株能稳定分泌抗OA BSA McAb的杂交瘤细胞株,竞争抑制法进一步证明了该抗体是特异针对OA的,腹水诱生法制备了大量的McAb。竞争抑制ELISA线性范围为0.24～125?ng/mL,线性方程y=-0.113?2logx+0.901?6,相关系数r=0.98,最低检出浓度为0.24?ng/mL。样品的加标回收率为97.07%～107.83%。结论获得了分泌抗OA的McAb的杂交瘤细胞株,建立了OA检测的简单、灵敏、高效的ELISA检测方法。     

E1B缺陷性腺病毒对p53缺失性白血病细胞的作用

目的 研究Ｅ１Ｂ缺陷性腺病毒（ｄｌｌ５２０）对３种ｐ５３缺失性白血病细胞Ｋ５６２、Ｊｕｒｋａｔ、ＨＬ－６０的作用。方法　以病毒 空斑试验检测 ｄｌｌ５２０在上述３种宿主细胞中的复制情况,用锥虫蓝染色检测ｄｌｌ５２０的杀细胞作用。结果 ｄｌｌ５２０在三 种白血病细胞中的复制明显低于阳性对照组,且对白血病细胞没有显著的致死作用。结论ｄｌｌ５２０对上述三种血液恶性 细胞不能产生有效的抑制作用。