Basic study towords reconstitution of normal and leukemic human hemopoiesis in small monkey of common marmoset

为了建立临床前人类造血功能正常和白血病的猴模型，我们引进了普通绒猴小猴。1)正常和白血病人绒猴造血细胞的构建：我们在超声引导下，将CD34阳性的人脐血细胞移植到绒猴胎儿体内。到目前为止，我们已经将CD34阳性的人脐血细胞移植到8只母猴的16只胎儿体内。我们还将人白血病细胞BV173、IMS-Bc1或Meg-01移植到7只母猴的14只胎儿体内。在开始时，经历了堕胎和移植，但使用高灵敏度的超声检查技术已经得到了改进。2)常见绒猴淋巴细胞的特性：我们分离到的绒猴淋巴细胞组分呈人类CD4、CD8或CD56单抗阳性。人类CD4或CD8抗体阳性的淋巴细胞在绒猴中也被认为是有功能的。CD56阳性的淋巴细胞还需要进一步鉴定，以阐明其特征。迄今为止，我们已经证明了人类与普通绒猴的造血细胞和淋巴细胞的相似特征。这些发现支持用人/绒猴造血嵌合体构建绒猴的可能性。这类绒猴将非常有利于在临床前水平上研究新开发的基因转移载体和生物修饰剂的安全性和有效性。虽然我们还没有成功构建这样的嵌合体绒猴，但我们已经开发了几种新的策略，将使异种移植更加安全和有效。构建绒猴临床前模型系统以研究正常人和白血病患者的造血功能还需要进一步的实验。

For the purpose of constructing preclinical monkey models with normal and leukemic human hematopoiesis, we have introduced small monkey of common marmoset. The followings are the progress in this year.1) Construction of normal and leukemic human hematopoiesis in marmoset :We transplanted CD34 positive human umbilical cord blood cells intraperitoneally to marmoset fetus under the guide of ultrasonograph. We have so far transplanted the CD34 positive human cord blood cells to 16 fetuses in 8 mother marmosets. We have also transplanted human leukemia cells of BV173, IMS-BC1 or Meg-01 to 14 fetuses in 7 mother marmosets. In the beginning abortions followed by the transplantations were experienced but the techniques have been improved using highly sensitive ultrasonography. Now we are following up carefully the in vivo constitution of human/marmoset chimera hematopoiesis in newborn marmosets.2) Characterization of common marmoset lymphocytes : We have isolated marmoset lymphocyte fractions which were positive for human CD4, CD8 or CD56 monoclonal antibodies. The lymphocytes positive for human CD4 or CD8 antibody were considered also to be functional in marmosets. CD56 positive lymphocytes should furthermore be characterized to elucidate its characters.We have so far demonstrated the similar characters of hematopoietic cells and lymphocytes between human and common marmosets. These findings support the possibility of constructing marmoset with human/marmoset hematopoietic chimera. Such marmosets would be very beneficial to study the safety and efficacy of newly developed gene transfer vectors as well as biological modifiers at preclinical levels. Although we have not successfully constructed such chimera marmosets yet, we have developed several new strategies which would make the xenografting more safely and efficiently. Further experiment is required to construct marmoset preclinical model system to study normal and leukemic human hematopoiesis.