Lemoli R M, Tazzari P L, Fortuna A, Bolognesi A, Gulati S C, Stirpe F, Tura S
Istituto di Ematologia Seragnoli, Università di Bologna, Italy.
Bone Marrow Transplant. 1994 Apr;13(4):465-71.
Human CD34+ hematopoietic cells were purified using the avidin-biotin immunoabsorption technique. The selected population showed 78.6 +/- 3% CD34+ cells and the overall recovery of CD34+ cells, CFU-GM and BFU-E from the starting population was 34 +/- 5%, 71 +/- 4% and 67 +/- 2%, respectively. Hematopoietic progenitor cell purification also resulted in 3 log of normal T cell depletion from the bone marrow (BM) by immunofluorescence analysis. Moreover, when unseparated BM cells were mixed with the CD34- lymphoma cell lines D430B and Raji, the removal of greater than 3 log of tumor cells from the enriched CD34+ cell fraction was demonstrated. To increase the neoplastic cell purging, several immunotoxins (IT) containing the ribosome-inactivating protein (RIP) saporin and directed toward the lymphoid-associated antigens CD30 and CD2 were prepared. Our experiments showed only a minimal toxicity of the immunoconjugates on colony-forming cells (CFC) derived from purified CD34+ cells. Conversely, all the ITs were very effective in inhibiting protein synthesis and growth of normal and neoplastic lymphoid cells. Further experiments demonstrated that the sequential combination of CD34+ cells purification and IT treatment resulted in 5 or more log of tumor cell purging with no additional loss of BM progenitor cells.
使用抗生物素蛋白-生物素免疫吸附技术纯化人CD34+造血细胞。所选群体显示78.6±3%的CD34+细胞，从起始群体中CD34+细胞、CFU-GM和BFU-E的总体回收率分别为34±5%、71±4%和67±2%。通过免疫荧光分析，造血祖细胞纯化还导致骨髓（BM）中正常T细胞减少3个对数。此外，当未分离的BM细胞与CD34-淋巴瘤细胞系D430B和Raji混合时，从富集的CD34+细胞组分中去除了超过3个对数的肿瘤细胞。为了增加肿瘤细胞清除率，制备了几种含有核糖体失活蛋白（RIP）皂草素并靶向淋巴相关抗原CD30和CD2的免疫毒素（IT）。我们的实验表明，免疫偶联物对源自纯化CD34+细胞的集落形成细胞（CFC）只有最小的毒性。相反，所有的IT在抑制正常和肿瘤性淋巴细胞的蛋白质合成和生长方面都非常有效。进一步的实验表明，CD34+细胞纯化和IT治疗的顺序组合导致肿瘤细胞清除5个或更多对数，而BM祖细胞没有额外损失。
