抗人粒细胞单克隆抗体的特异性鉴定及其核素^(99)Tc^m标记

Identification and ^(99)Tc^m labelling of a monoclonal antibody against human granulocytes

目的以人粒细胞免疫Balb/c小鼠制备抗粒细胞单克隆抗体(mAb),并对其组织特异性进行鉴定,同时评价其在兔炎症模型中的显像价值。方法常规免疫制备抗粒细胞mAb经间接ELISA法、流式细胞仪和SP免疫组化进行特异性鉴定。99Tcm间接法标记SZ102。兔左下肢炎症模型制备完毕,耳缘静脉注入99TcmSZ1024h后探讨其在兔炎症模型中的体内分布。结果制备抗粒细胞mAbSZ102,其免疫球蛋白亚类为IgG1。流式细胞仪测定表明其与外周血粒细胞、单核细胞呈强阳性反应,而与淋巴、红细胞、血小板不反应,并且与正常骨髓中较成熟粒细胞、单核前体起反应。SP免疫组化法检测正常人组织中SZ102抗原分布,显示抗原表达在肝、肺、脾、胸腺、淋巴结组织中的巨噬细胞表面。注射99TcmSZ1024h后,离体炎症肌肉/血液和对侧正常肌肉/血液单位质量放射性比值分别为0.22±0.02和0.02±0.01。结论SZ102是一株稳定高效价抗粒细胞杂交瘤细胞株,99TcmSZ102具有活体内炎症定位导向能力,对隐匿性炎症的放免显像可能有一定的诊断价值。

Objective To prepare a mAb （SZ-102） against granulocytes, identify its characteristics, and evaluate its role in the method of ^99Tc^m labelling for the detection of inflammatory areas in rabbits. Methods Balb/c mouse was immunized through intraperitional route. The mAb was screened by indirect ELISA and the tissue specificity of the mAb was identified by flow cytometry and SP staining. The mAb SZ-102 was indirectly labelled with 99Tcm , and then injected into the rabbit experimental models of inflammatory areas. The distribution of ^99Tc^m-SZ-102 in rabbit was determined at 4 h postinjection. Results The mAb SZ-102 against grantdocytes was obtained and its Ig subclass was IgG1. The mAb SZ-102 could react with human grantdocytes, monocytes, and their bone marrow precursors, but not with human lymphocytes or red blood cells. SZ-102 antigen was expressed on the macrophage of liver, lung, thymus gland, spleen, and lymph node. The ratio of inflammatory muscle to blood was 0.22 ±0.02, while the ratio of normal muscle to blood was 0.02± 0.01 at 4 h postinjection of 9^99Tc^m-SZ-102. Conclusion One mAb SZ-102 against granulecytes with higher immunoactivity and tissue specificity will be of certain value for detecting inflammatory areas in clinically concealed infectious diseases.