抗人CD100单克隆抗体的制备与初步鉴定

目的：研制可识别天然CD100分子的单克隆抗体（mAb)。方法：采用真核表达的CD100分子免疫BALB/c小鼠，以间接ELISA法筛选阳性杂交瘤，并分别用人急性T淋巴细胞性白血病细胞系MOLT-4，猿T淋巴细胞系MLA-144及猴EB病毒转化的B淋巴细胞系（BLCL），进行间接免疫荧光染色和流式细胞术鉴定mAb的特异性。结果：共获得7株可稳定分泌mAb的杂交瘤细胞株。所有mAb均可识别人和猴细胞系的膜表面天然CD100分子，其中6株可识别猿细胞系膜表面的天然CD100分子，结论：成功地制备了7株抗CD100分子的特异性mAb。为研究人和猿，猴CD100分子的结构与功能提供了新的手段。

Preparation and identification of monoclonal antibodies against CD100 molecule

AIM: To prepare and identify the monoclonal antibodies (mAbs) against CD100. METHODS: BALB/c mice were immunized with eukaryotic expressed CD100 protein. The splenocytes from immunized mice were fused with Sp2/0 cells and the positive hybridoma clones were screened by indirect ELISA and mAbs reacting to GD100 antigen were identified by indirect immunofluorescent staining and flow cytometry with human acute T lymphoblastic leukemia cell line MOLT-4, activated human PBMC, gibbon T lymphocyte cell line MLA-144 and monkey EBV-transformed B lympho-blast cell line( BLCL). RESULTS: Seven hybridomas secreting mAbs against CD100 molecule were obtained. All the mAbs could react to natural CD100 expressed on human MOLT-4 and activated PBMCs as well as monkey BLCL cells, and six out of them could react to gibbon MLA-144 cells. CONCLUSION; A set of mAbs recognizing CD100 molecule expressed on human, monkey and gibbon cells are obtained, which may provide an useful tool for studying the structure-function relationship of GD100 molecule of human, as well as gibbon and monkey.