Hematopoietic activity of common marmoset CD34 cells isolated by a novel monoclonal antibody MA24
Hematopoietic activity of common marmoset CD34 cells isolated by a novel monoclonal antibody MA24
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DOI：
10.1016/j.exphem.2004.06.007
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发表时间：
2004-09-01
影响因子：
2.6
通讯作者：
Asano, S
中科院分区：
文献类型：
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作者：
Izawa, K;Tani, K;Asano, S
目标。我们以一种小的新大陆猴子，普通的绒猴(Callithrix Jacchus)为研究对象，建立了一种治疗血液疾病的非人类灵长类动物模型。在本研究中，我们研制了第一批抗绒猴CD34的单抗，并用其中一种单抗分离的细胞群体进行了体外和体内造血活性的检测。利用逆转录聚合酶链式反应和快速扩增cDNA端的方法，从骨髓来源的RNA中克隆了编码人CD34同源基因的绒猴基因。绒猴CD34的氨基酸序列与人类的同源性为81%。制备了5株抗绒猴CD34转基因细胞的鼠单抗。一种具有代表性的单抗MA24(IgM)与大约0.5%~1%的骨髓单个核细胞(MNC)发生反应，其中集落形成单位粒细胞/巨噬细胞(CFU-GM)的浓度大约是整个BM单个核细胞的11-75倍。异种移植1个月后，流式细胞仪检测证实NOD/SCID小鼠绒毛CD34(+)细胞向多系分化。这些结果表明MA24可用于临床前实验的绒猴模型中造血祖细胞的分析和浓缩。(C)2004年国际实验血液学学会。由爱思唯尔公司出版。
Objective. We focused on a small New World monkey, the common marmoset (Callithrix jacchus), to establish a nonhuman primate model of the treatment of hematological disorders. In this study, we developed the first monoclonal antibodies (MAbs) against marmoset CD34 and tested the in vitro and in vivo hemopoietic activity of cell populations isolated using one of these MAbs.Methods and Results. Marmoset cDNA encoding a human CD34 homologue was cloned from bone marrow (BM)-derived RNA using reverse transcription polymerase chain reaction and rapid amplification of cDNA ends. The amino acid sequence of the marmoset CD34 had 81% homology with the human sequence. Five mouse MAbs were raised against marmoset CD34 transfectant. One representative MAb, MA24 (IgM), reacted with approximately 0.5 to 1% of BM mononuclear cells (MNCs), where the colony-forming unit granulocyte/macrophage (CFU-GM) was enriched approximately 11- to 75-fold as compared with the whole BM MNCs. Multilineage differentiation of marmoset CD34(+) cells in NOD/SCID mice was confirmed by flow cytometry 1 month after xenotransplantation.Conclusion. These results demonstrated that MA24 is useful for the analysis and enrichment of hematopoietic progenitor cells in the marmoset model for preclinical experiments. (C) 2004 International Society for Experimental Hematology. Published by Elsevier Inc.