Effects of PBSC reinfusion on tumor immunity in patients with urologic malignancies

外周血祖细胞（PBSC）回输促进了泌尿系肿瘤和血液系统恶性肿瘤患者清髓性化疗的进展。在人骨髓和外周血中作为祖细胞的实际集落形成细胞可以通过CD 34抗原的表达来鉴定。目前普遍认为，回输的外周血干细胞中CD 34阳性细胞的数量与造血恢复有关。使用流式细胞术分析，首先我们建立了用于检测外周血和PBSC收获细胞中的CD 34阳性细胞浓度的技术（发表在Biotrapy上）。我们的检测CD 34阳性细胞的技术已经很好地建立，并与集落形成试验的结果（发表在Igakukensa）相关。另一方面，PBSC收获的细胞含有大量的巨噬细胞。近年来研究发现，巨噬细胞能强烈分泌细胞因子刺激造血。第二部分研究外周血干细胞回输对患者免疫功能的调节作用。CD 69抗原是一种早期活化抗原，也是NK-基因家族复合物相关抗原，通过双色流式细胞术分析PBSC收获细胞（ISBT）中细胞因子分泌的循环单核细胞。我们证实CD 69抗原表达的变化是PBSC动员（ISBT）的良好指标。然而，在这个免疫学问题变得清晰之前，似乎还需要进一步的研究。总之，我们建立了检测外周血和PBSC收获细胞中CD 34阳性细胞的技术。外周血单核细胞表面CD 69抗原表达的改变可能反映了患者的免疫反应。

Peripheral blood progenitor cells (PBSC) reinfusion has contributed to the advance of myeloablative chemotherapy in patients with urologic cancers as well as hematological malignancies. Virtually colony forming cells as progenitors in human bone marrow and peripheral blood can be identified by the expression of the CD34 antigen. It is commonly believed that number of CD34 positive cells in PBSC reinfusion have been correlated with hematopoieticrestore. Using flow cytometric analysis, first we established a technique for detecting CD34 positive cells' concentration in peripheral blood and PBSC harvesting cells (published in Biothrapy). Our technique for detecting CD34 positive cells had been well established and correlated with the results of colony formation assay (published in Igakukensa).On the other hands, PBSC harvested cells contained a number of macrophages. It was recently reported the such macrophages strongly secleted cytokines for stimulating hematopoiesis. We secondly investigated on the effect of PBSC reinfusion to the immune-modulation in patients. CD69 antigen that is an early activating antigen and also an antigen related with NK-gene family complex was identified by dual color flow cytometric analysis on cytokine seclated circulating monocytes in PBSC harvesting cells (ISBT). We confirmed change in CD69 antigen expression to be a good indicator of PBSC moblization (ISBT). However, before this immunological issue become clear, it seemed further investigations will be required. In conclusion, our establishment technique for detecting CD34 positive cells in peripheral blood and PBSC harvesting cells is well established. Change in CD69 antigen expression on circulated monocytes might demonstrate some immunological reaction in patients.