通过新型单克隆抗体MA24分离的普通狨猴CD34细胞的造血活性。

Hematopoietic activity of common marmoset CD34 cells isolated by a novel monoclonal antibody MA24.

作者信息

Izawa Kiyoko, Tani Kenzaburo, Nakazaki Yukoh, Hibino Hitoshi, Sugiyama Hajime, Kawasaki Akemi, Sasaki Erika, Nishioka Chieko, Ishii Hajime, Soda Yasushi, Yagita Hideo, Tanioka Yoshikuni, Tojo Arinobu, Asano Shigetaka

机构信息

Institute of Medical Science, University of Tokyo, Tokyo, Japan.

出版信息

Exp Hematol. 2004 Sep;32(9):843-51. doi: 10.1016/j.exphem.2004.06.007.

DOI:10.1016/j.exphem.2004.06.007

PMID:15345286

Abstract

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.

摘要

目的

我们聚焦于一种小型新大陆猴——普通狨猴（Callithrix jacchus），以建立一种治疗血液系统疾病的非人灵长类动物模型。在本研究中，我们制备了首个针对狨猴CD34的单克隆抗体（MAb），并测试了使用其中一种MAb分离的细胞群体的体外和体内造血活性。

方法与结果

使用逆转录聚合酶链反应和cDNA末端快速扩增技术，从骨髓（BM）来源的RNA中克隆编码人CD34同源物的狨猴cDNA。狨猴CD34的氨基酸序列与人类序列具有81%的同源性。制备了5种针对狨猴CD34转染子的小鼠单克隆抗体。一种代表性的单克隆抗体MA24（IgM）与约0.5%至1%的BM单个核细胞（MNC）发生反应，其中粒细胞/巨噬细胞集落形成单位（CFU-GM）与整个BM MNC相比富集了约11至75倍。异种移植1个月后，通过流式细胞术证实了狨猴CD34+细胞在NOD/SCID小鼠中的多系分化。