抗人粒细胞单克隆抗体的研制和鉴定及其放射免疫显像

Preparation and Radioimmunoimaging of a Monoclonal Antibody Against Human Granulocytes

以人粒细胞常规免疫和融合BALB/c小鼠 ,建立一株稳定的高效价的抗人粒细胞单克隆抗体杂交瘤细胞株SZ 10 2 ,并对其组织特异性进行鉴定及探讨在炎症模型显像中的潜在价值。结果表明 ,McAbSZ 10 2免疫球蛋白亚类为IgG1。流式细胞术测定显示其与外周血液中的粒细胞和单核细胞呈强阳性反应 ,而与淋巴细胞、红细胞和血小板不起反应 ,但与正常人骨髓中的较成熟的粒细胞、单核前体细胞发生反应。SP免疫组化法检测正常人组织切片中SZ 10 2抗原阳性细胞分布的结果显示 ,该抗原主要表达在肝、肺、脾、胸腺、淋巴结组织中的巨噬细胞表面。以亚氨基噻吩 (2 IT)修饰 ,99Tcm 标记葡庚糖酸钠 (GH)配体交换法标记SZ 10 2。给具有左下肢炎症模型的家兔 ,由耳缘静脉注入99Tcm 抗人粒细胞单克隆抗体SZ 10 2进行SPECT显像 ,以99Tcm 标记非特异性鼠IgG作为阴性对照 ,结果表明99Tcm SZ 10 2在家兔炎症部位显像清晰 ,其最佳显像时间为注射后 2 - 4小时 ,而99Tcm 标记非特异性鼠IgG在家兔炎症部位未能清晰显像。结论 :99Tcm SZ 10 2具有活体内炎症定位导向能力 ,显像时间短 。

The aim of this research is to prepare a novel monoclonal antibody against granulocytes by the intraperitional routine procedure and evaluate the usefulness of 99 Tc m labelled anti granulocyte monoclonal antibody (McAb) SZ 102 for the detection of experimental inflammatory areas in rabbits. It was characterized as IgG1 subclass by the double immunodiffusion analysis. The flow cytometry demonstrated that SZ 102 reacted selectively with human granulocytes, monocytes, and with their bone marrow precursors, while human lymphocytes, red blood cells and platelets remained negative. In addition, SZ 102 antigen was expressed on the macrophages of liver, lung, thymus, spleen and lymph node by immunohistochemical SP methods. It is suggested that McAb SZ 102 is mainly against granulocytes. SZ 102 was labelled with 99 Tc m using 2 iminothiolane modification McAb and 99 Tc m glucoheptonate(GH) transchelation method. The experimental rabbit model of inflammatory areas was prepared,through injecting with 99 Tc m SZ 102 by ear edge vein, and then imaged by SPECT. 99 Tc m labelled murine IgG was used as a negative control. The inflammatory areas in rabbits were clearly imaged at 2 to 4 hour after injection of 99 Tc m SZ 102, while the control group after injection of 99 Tc m labeled murine IgG was negative. In conclusion, the McAb SZ 102 may be a potential agent for the diagnosis and localization of inflammatory areas of carcinomas and clinically concealed infectious diseases.