医学毕业论文白细胞新单克隆抗体zch-2b8a的表达谱分析及其意义

XX大学 毕业论文 白细胞新单克隆抗体ZCH-2B8a的表达谱分析及其意义 姓 名 2014年6月25日 白细胞新单克隆抗体ZCH-2B8a的表达谱分析及其意义 作者汤永民郭莉，杨世隆，沈红强，钱柏芹，张怡，张海忠 【摘要】ZCH-2B8a （IgG2a ）是作者研究室最近采用髓细胞性白血病细胞系 KGla作为免疫原自行研制的鼠抗人造血细胞分化抗原的单克隆抗体，已提交第 8届国际人类白细胞分化抗原协作组会议（HLDA8）鉴定。根据HLDA8总部反 馈信息表明，该抗体识别的抗原性质不明，后者属国际上尚未认识的血细胞膜新 的分化抗原。本研究旨在分析ZCH-2B8a单克隆抗体与正常人外周血细胞成分、 骨髓及G-CSF动员的外周血CD34造血干/祖细胞和血液肿瘤细胞系的反应规律 及其意义。采用多参数流式细胞术分析2B8a抗体与正常及恶性肿瘤细胞表面 的反应性，每种细胞成分重复实验3次，以阳性细胞数之20为阳性。结果表明 2B8a抗原在外周血B细胞上表达（3/3例，平均阳性细胞数为26.29 ）,而在 T淋巴细胞和NK细胞上不表达（0/3例）；在粒细胞和单核细胞上阳性表达均为 2/3例，平均阳性细胞数分别是23.72 和59.84 ；在DC细胞、红细胞和血 小板上均不表达（0/3例）o2B8a抗原在骨髓CD34 细胞上的阳性表达是3/3例， 平均阳性细胞数39.33 ,而在G-CSF动员的外周血CD34细胞上的阳性表达 仅1/3例，平均阳性细胞数为1.25 。2B8a抗原在B系细胞系Raji、SMS-SB、 Nalm-6和Nall-1上的平均阳性细胞数分别为98.78 、98.61 、94.93 和 5.68 ；在T系细胞系Molt-3 的平均阳性细胞数为31.40 ,而在Molt-4、 JM和CCRF-CEM细胞上不表达；在髓系细胞系U937、Meg-0k HL-60、K562、 KGla和HEL92.1.7的平均阳性细胞数分别为67.78 、33.40 、29.70 、 28.19 、16.23 和8.02 ；在神经母细胞瘤细胞系 SK-N-SH、KCNR、BE、 LAN-1和SK-N-AS细胞以及结肠癌细胞系HR8348细胞上均不表达，而在羊膜 细胞系FL细胞上呈一定的阳性表达，平均阳性细胞数为45.03。结论在正 常外周血，2B8a抗体主要与B淋巴细胞和单核巨噬细胞反应，在骨髓和外周血 CD34造血干/祖细胞中也有不同程度的反应性；在细胞系上，2B8a抗体主要与 B系细胞系、单核巨噬细胞系及上皮性肿瘤细胞系反应，提示该抗体有可能用于 上述肿瘤的免疫学诊断与治疗。 病，淋巴瘤 Reactivity of a Novel Monoclonal Antibody ZCH-2B8a on Normal Hematopoietic Cells and Malignant Cell Lines and Its Significance AbstractsZCH-2B8a IgG2a is a novel monoclonal antibody McAb generated in laboratory of Children Hospital of Medical College, Zhejiang University recently using human myeloblastic leukemia cell line KGla as immunogen. This antibody has been ted to the 8th International Workshop and Conference on Human Leukocyte Differentiation Antigens HLDA8 and the results showed that the antibody recognized an unknown molecule on the surface of some blood cells. The aim of this study was to investigate the reactivity of this antibody on normal blood cells and malignant cell lines and to explore its possible application in clinical practice. The multi-parameter flow cytometry was used to analyze the expression pattern of 2B8a antigen in triplicate on normal blood components including T cells, B cells , natural killers NK, neutrophils, monocytes, dendritic cells DC, red blood cells RBC, platelets Pit, hematopoietic stem/progenitor cells derived from either bone marrow or G-CSF mobilized peripheral blood CD34 cells and malignant cell lines including 14 hematopoietic, 5 neuroblastoma, 1 colon cancer and 1 amniotic epithelium cell lines. The amount of positive cells 20 was considered as positivity. The results showed that 2B8a antibody reacted to 3/3 specimens of blood B cells with a positive rate of 26.29 and 2/3 specimens of monocytes with an average positive rate of 59.84. 2B8a was weakly reactive to neutrophils 23.72 and negative for T cells, NK, DC, RBC and Pit. The antibody reacted to all 3 marrow CD34 cells with an average positive rate of 39.33 while it was negative for G-CSF-mobilized CD34 peripheral blood stem/progenitor cells PBSC, 1.25. Cell line analysis showed that the an