Development of human disease model system using small nonhuman primate

(1)普通狨猴与人免疫血液学比较；使用抗人淋巴细胞抗体（anti - human lymphocyte Ab），采用MACS珠法分离T细胞。使用市售的MoAbs， T细胞的纯化效率高于B细胞。分离的T细胞，对抗人类CD4或CD8抗体有反应，被发现实际上是被丝裂原刺激的。(2)普通狨猴多能干细胞分析；我们克隆了狨猴CD34和c-kit的cdna。核苷酸序列分析表明，前者与人类对应序列的同源性分别为89%和95%。我们一直在生产抗绒猴CD34和c-kit单克隆抗体来分离绒猴多能造血干细胞。(3)绒猴主要组织相容性复合体（MHC）分析；利用我们开发的DNA分型方法，在狨猴MHCⅱ类DRB1等位基因中发现了7个新的等位基因。进一步，我们分析了I类等位基因，并将其定位在狨猴4号染色体上。(4)人体疾病模型的制作；我们制作了注射胶原蛋白后的狨猴关节炎模型。构建了LacZ和IL-13腺病毒载体。关节内注射腺病毒提示IL-13腺病毒抑制狨猴关节炎的有效性。我们还利用BCR/ABL逆转录病毒在普通狨猴中构建了人类白血病。我们一直在安全地进行实验，没有产生任何具有复制能力的逆转录病毒。

(1) Immuno-hematological comparison between common marmoset and human ; T cell fraction was isolated by MACS beads method using antihuman lymphocyte Ab. The purification efficacy was higher for T cells than B cells using commercially available MoAbs. The isolated T cells, reactive to antihuman CD4 or CD8 Abs, found to be actually stimulated by mitogen.(2) Analysis of common marmoset multipotent stem cells ; We have cloned cDNAs for marmoset CD34 and c-kit. Nucleotide sequence analysis showed that the former had 89% and the latter had 95% homologies respectively with human counterpart sequences. We have been producing antimarmoset CD34 and c-kit monocloncal Ab to isolate marmoset multipotent hematopoietic stem cells.(3) Analysis of marmoset major hisocompatibility complex (MHC) ; We have found 7 new alleles in marmoset MHC class II DRB1 alleles using DNA typing method which we developed. Furthermore, we analyzed class I alleles and localized the genes to marmoset chromosome 4.(4) Production of human disease model ; We have produced marmoset arthritis model after collagen injection. We constructed LacZ and IL-13 adenoviral vectors. The intraarticular injection of the adenovirus suggested the usefulness of IL-13 adenovirus to suppress arthritis in marmosets. We also have been constructing human leukemia in common marmoset using BCR/ABL retrovirus. We have been performing the experiment safely, without producing any replication competent retrovirus.