抗毒死蜱单克隆抗体的研制

用与牛血清蛋白(BSA)交联的毒死蜱人工抗原(CHBu-BSA)免疫的BALB/C鼠脾细胞与SP2/0鼠骨髓瘤细胞融合,经筛选和克隆,得到了1株能稳定分泌毒死蜱抗体的单克隆细胞株(1G10),1G10 的抗体类型及亚类均为IgG1, 其轻链为κ链.制备单克隆抗体腹水,腹水的间接ELISA效价在1×10-6以上.该单克隆抗体与毒死蜱特异性结合反应的50%抑制质量浓度为80.8 μg·L-1,与其他毒死蜱结构类似物无交叉反应.     

A monoclonal antibody to limbic system neurons

A monoclonal antibody produced against hippocampal cell membranes labeled the surface of neurons in the rat limbic system. With a few exceptions, all nonlimbic components were unstained. This specific distribution of immunopositive neurons provides strong evidence of molecular specificity among functionally related neurons in the mammalian brain and supports the concept of a limbic system.     

2种植物多糖对PRRSV灭活苗免疫猪抗体及T淋巴细胞亚群的影响

将24头35日龄断奶仔猪随机分为6组,2种多糖(山药多糖和黄芪多糖)分别以高、低2个剂量(山药多糖剂量17.10、8.55mg/kg,黄芪多糖剂量为11.50、5.75mg/kg)和猪PRRSV灭活苗同时注射,于免疫后不同时间点采血,监测PRRSV抗体水平和外周血中T淋巴细胞亚群的变化.结果表明,2种多糖均能增强免疫猪的体液免疫,其中以高剂量的黄芪多糖效果较好;2种多糖均能促进猪CD3+细胞的增殖,其中以山药多糖效果较好,提示2种多糖对PRRSV灭活苗免疫效果均有较好的增强作用.     

Genetic control of the antibody response: relationship between immune response and histocompatibility (H-2) type

The immune responses of inbred mice to a related series of three synthetic polypeptide antigens are genetically controlled traits which are closely correlated with the genotype for the major histocompatibility (H-2) locus. All strains of the same H-2 type exhibit the same pattern of immune response, independent of the remainder of a given strain's genetic background. There is marked antigen-specific polymorphism between strains of different H-2 types with respect to their patterns of response.     

环丙沙星单克隆抗体的制备

【目的】制备环丙沙星(Crprofloxacin,CPFX)单克隆抗体(monoclonal antibody,mAb),并对其免疫学特性进行鉴定,为畜产品CPFX残留的快速检测提供技术支持。【方法】采用碳二亚胺法(EDC),将载体蛋白牛血清白蛋白(Bovine serum albumin,BSA)和鸡卵白蛋白(ovalbumin,OVA)分别与CPFX偶联合成免疫抗原CPFX-BSA和包被抗原CPFX-OVA,经紫外(UV)扫描和聚丙烯酰胺凝胶电泳(SDS-PAGE)鉴定后免疫BALB/c小鼠,应用细胞融合技术筛选分泌CPFX mAb的杂交瘤细胞株,采用体内诱生腹水法制备CPFX mAb,并对其染色体核型、效价、敏感性和特异性等免疫学特性进行鉴定。【结果】UV和SDS-PAGE鉴定结果表明,人工抗原偶联成功;免疫的5只小鼠血清抗体效价均达到了1∶105左右,其中4号小鼠血清CPFX抑制效价较高且IC50最低(17.21μg/L),融合后筛选出1株敏感特异的杂交瘤细胞,命名为3D2。3D2细胞培养上清效价为1∶(1.28×103),腹水效价为1∶(1.6×105)。CPFX mAb对CPFX的IC50为9.95μg/L,对恩诺沙星的交叉反应率为128.39%,与其他同系的喹诺酮类抗生素的交叉反应率很低。【结论】获得了高效价、敏感、特异的抗CPFX mAb,为CPFX残留的快速检测奠定了技术基础。     

人类免疫缺陷病毒单克隆抗体在小麦胚乳中的表达

为了建立利用小麦生产抗人类免疫缺陷病毒(HIV)单克隆抗体的方法,将植物胚乳特异启动子AsGlo分别与该抗体的重链(HC)和轻链(LC)连接构建小麦表达载体,以基因枪法转入小麦品种扬麦158。转基因T0～T3代PCR鉴定表明,HC和LC在小麦基因组中共整合和稳定遗传。RT-PCR分析表明,IgG重链和轻链基因可在小麦胚乳中正常转录。酶联免疫吸附(ELISA)进一步证实了IgG在小麦胚乳中的积累和抗原结合活性,表明IgG蛋白能够在小麦胚乳中稳定表达,可利用小麦种子生产抗HIV抗体。     

抗鸭IL-2单克隆抗体的制备与鉴定

用纯化的重组鸭IL-2蛋白免疫BALB/c 小鼠,取其脾细胞与SP2/0骨髓瘤细胞融合,对杂交瘤细胞进行筛选,阳性孔经三次有限稀释法克隆,成功获得5株能稳定传代并分泌抗鸭IL-2单克隆抗体(mAb)的杂交瘤细胞,分别命名为:1H4、2B3、4G12、5F6、5H6,其中一株对鸡IL-2具有交叉反应性.各单克隆抗体腹水间接ELISA效价在1∶32000～1∶512000之间,抗体类型除mAb 5H6为IgG2a亚类外,其余4株mAb均为IgG1亚类,5株mAb的轻链都为κ链.经Western-blot分析表明,所获得的5株抗鸭IL-2单克隆抗体与鸭IL-2具有反应性,推测它们是针对序列决定簇的.     

Genetics of the antibody response to dextran in mice

The immune response to dextran having the alpha-1,3 linkage may be under the control of antibody structural genes. Mice that respond well to this antigen produce antibody restricted with respect to light chain class (lambda) and to an antigenic determinant resulting from a particular heavy and light chain interaction. The response to dextran is controlled by a locus linked to the-heavy chain locus.     

抗家蚕浓核症病毒单克隆抗体实用化研究

制备了家蚕浓核症诊断试剂盒,将单克隆抗体与双向免疫技术相结合,用于家蚕浓核症的诊断.采用此试剂盒,对家蚕浓核症的最早检出时间为感染病毒后48小时.若将此单克隆抗体与免疫酶技术或萤光抗体技术结合,则能在感染病毒后12～16小时的家蚕中肠组织内检测到病毒抗原.配制的单克隆抗体应用液效价稳定,在37℃下放置一个月,室温下放置两个月仍然有效.单克隆抗体能代替常规抗血清用于家蚕浓核症的诊断.     

抗重组鸡IL-2单克隆抗体的制备与鉴定

以纯化的重组鸡IL-2蛋白(rchIL-2)为免疫原,按常规方法免疫BALB/c小鼠,取其脾细胞与SP2/0骨髓瘤细胞融合,对杂交瘤细胞进行筛选,阳性孔经3次有限稀释法克隆,最终获得4株持续且稳定分泌抗鸡IL-2单克隆抗体(MAb)的杂交瘤细胞株,分别命名为:1A12、4A3、3E7和4E1.4株MAb的抗体类型均为IgG1,轻链为κ;MAb的ELISA效价分别为5.12×105、1.02×106、2.56×105和4.09×106.Western-blot分析表明,所获得的1A12、4A3、3E7和4E1 4株单克隆抗体与rchIL-2均有反应.MAb的制备成功为进一步开展chIL-2的功能研究奠定了基础.     

抗鸡传染性支气管炎病毒单克隆抗体的制备与应用

将鸡传染性支气管炎病毒M41 株尿囊液差速离心纯化 ,制备抗原 ;应用杂交瘤技术建立了 3株分泌抗鸡传染性支气管炎病毒的杂交瘤细胞株 ,其免疫球蛋白亚类分别属于IgG1 、IgG2a、IgG2b。 3株单克隆抗体与鸡新城疫病毒、传染性法氏囊病病毒、产蛋下降综合征病毒及禽流感病毒均无交叉反应 ,YNZ - 55和YNZ - 62可识别多数鸡传染性支气管炎标准毒株及地方分离株 ,其腹水ELISA效价达 1 0 - 5,且识别的抗原位点互不重叠 ,为高亲和力的群特异性单克隆抗体 ,是制备鸡传染性支气管炎快速诊断产品的理想试剂     

Herpes simplex virus glycoprotein D: human monoclonal antibody produced by bone marrow cell line

Normal bone marrow cells from a donor positive for herpes simplex virus were transformed with Epstein-Barr virus. The resulting lymphoblastoid cell line has secreted immunoglobulin G1 of the kappa type continuously for 2 years. This immunoglobulin, detected both on the cell surface and in the cytoplasm, reacts with cells infected with herpes simplex virus. It defines an antigen that comigrates with the 55-kilodalton glycoprotein D of herpes simplex virus type 1 and neutralizes the infectivity of herpes simplex viruses 1 and 2.     

蜱诱导宿主免疫的研究进展

蜱对人畜的危害很大,传统灭蜱措施主要是化药控制,而耐药性的出现,使得人们转而使用新的方法控制蜱。本文介绍了利用免疫方法控制蜱的一些成就,并对该研究方向做了进一步展望。     

Stress-induced modulation of the immune response

After mice were exposed to a daily auditory stressor for varying lengths of time, the responses of their splenic lymphoid cells in vitro were assessed. Both the blastogenic activity of concanavalin A or lipopolysaccharide and the ability of immune lymphocytes to lyse P815 target cells showed the same patterns of immunosuppression and enhancement.     

睾酮的单克隆抗体酶免疫分析法研究

利用睾酮单克隆抗体,用非平衡法建立鉴定了睾酮的固相微滴板酶免疫分析法,整个测试过程在1小时内完成,结合率(E/E_0)与睾酮浓度的自然对数(InT)之间典型的直线回归方程式为:E/E_0=162.29 InT—19.45(r=-0.9783).用建立的方法测定了海佩科肉鸡59日龄血清睾酮浓度,公鸡为82.08±30.53ng/DL,母鸡为58.84±14.45ng/DL,肉鸡血清睾酮浓度与其体重存在显著的正相关(r=0.4002,n=32).     

H3N2亚型SIV血凝蛋白单克隆抗体的制备

【目的】制备H3N2亚型猪流感病毒(SIV)的单克隆抗体,为SIV的鉴别诊断奠定基础。【方法】将A/swine/Henan/1/2010(H3N2)猪流感病毒初步浓缩后,免疫6周龄的BALB/c小鼠,取小鼠脾细胞与瘤细胞NS0融合,采用间接ELISA方法筛选杂交瘤细胞,对杂交瘤细胞分泌的单克隆抗体进行Western blot检验和抗原结合活性检测。【结果】经过3次有限稀释法克隆纯化,最终得到1株能稳定分泌单克隆抗体的阳性细胞株,命名为1C10,经检测其腹水抗体效价为1∶51 200。单克隆抗体亚型鉴定结果表明,1C10为IgG1亚型,轻链类型为κ链。Western blot检测结果表明,这株单克隆抗体能与H3N2特异性结合,而且能特异性识别血凝素蛋白(HA),而不与H1N1亚型猪流感病毒、伪狂犬病毒、猪圆环病毒2型、猪细小病毒发生交叉反应。【结论】制备获得1株抗猪流感H3N2病毒HA蛋白的单克隆抗体,可用于猪流感病毒鉴别诊断方法的建立。     

Reversal of experimental allergic encephalomyelitis with monoclonal antibody to a T-cell subset marker

Administration of a monoclonal antibody (GK1.5) that recognizes the L3T4 marker present on helper T cells prevented the development of experimental allergic encephalomyelitis (EAE) in mice. Furthermore, treatment with GK1.5 reversed EAE when the antibody was given to paralyzed animals. In vivo injection of GK1.5 selectively reduced the number of L3T4+ cells in the spleen and the lymph nodes. These results suggest that manipulation of the human equivalent of the murine L3T4+ T-cell subset with monoclonal antibodies may provide effective therapy for certain autoimmune diseases.