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ZCH-2B8a(IgG2a)是作者研究室最近采用髓细胞性白血病细胞系KG1a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HL-DA8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≥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)。2B8a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上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-01、HL-60、K562、KG1a和HEL92.1.7上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。
ZCH-2B8a (IgG2a) is a monoclonal antibody recently developed by the author laboratory for the use of myeloid leukemia cell line KG1a as a self-developed mouse anti-human hematopoietic differentiation antigen and has been submitted to the 8th International Symposium on Human Leukocyte Differentiation Antigen ( HLDA8) identification. According to the feedback information of HL-DA8 headquarters, the antigen recognized by the antibody is unknown in nature, and the latter belongs to a new differentiated antigen of the blood cell membrane which is not yet known in the world. The purpose of this study was to analyze the relationship between ZCH-2B8a monoclonal antibody and normal human peripheral blood cells, bone marrow and G-CSF-mobilized peripheral blood CD34 + hematopoietic stem / progenitor cells and hematological tumor cell lines. The multi-parameter flow cytometry was used to analyze the reactivity of 2B8a antibody with the surface of normal and malignant tumor cells. The experiment was repeated 3 times for each cell component, and the positive cells were ≥20% positive. The results showed that the expression of 2B8a antigen on peripheral blood B cells (3/3 cases, the average number of positive cells was 26.29%), but not in T lymphocytes and NK cells (0/3 cases); in granulocytes and single The number of positive cells on nucleus was 2/3, and the average number of positive cells was 23.72% and 59.84% respectively. No expression was found on DCs, erythrocytes and platelets (0/3). The positive expression rate of 2B8a antigen in bone marrow CD34 + cells was 3/3 and the average number of positive cells was 39.33%. The positive expression of 2B8a antigen was only 1/3 in CD34 + cells mobilized from G-CSF, and the average number of positive cells was 1.25 %. The average number of positive cells of 2B8a antigen in the B lines Raji, SMS-SB, Nalm-6 and Nall-1 were 98.78%, 98.61%, 94.93% and 5.68% respectively. On the T cell line Molt-3 The average number of positive cells was 31.40%, but not on Molt-4, JM and CCRF-CEM cells; the average positive expression on myeloid cell lines U937, Meg-01, HL-60, K562, KG1a and HEL92.1.7 The cell numbers were 67.78%, 33.40%, 29.70%, 28.19%, 16.23% and 8.02%, respectively. In neuroblastoma cell lines SK-N-SH, KCNR, BE, LAN-1 and SK- Colon cancer cell line HR8348 cells were not expressed, but in the amniotic cell line FL cells showed a certain positive expression, the average number of positive cells was 45.03%. Conclusions: In normal peripheral blood, 2B8a antibody mainly reacts with B lymphocytes and monocyte-macrophages, and also has different degrees of reactivity in bone marrow and peripheral blood CD34 + hematopoietic stem / progenitor cells. On the cell lines, 2B8a antibody mainly interacts with B cell line, monocyte-macrophage cell line and epithelial tumor cell line, suggesting that the antibody may be used for the above immunological diagnosis and treatment of tumors.