Lee T H, Paglieroni T, Ohto H, Holland P V, Busch M P
Research and Scientific Services, Blood Centers of the Pacific (formerly Irwin Memorial Blood Centers), San Francisco, CA, USA.
Blood. 1999 May 1;93(9):3127-39.
We recently reported detection of a transient increase in circulating donor leukocytes (WBCs) in immunocompetent recipients 3 to 5 days posttransfusion (tx) (Blood 85:1207, 1995). We have now characterized survival kinetics of specific donor WBC subsets in additional tx populations. Eight female elective surgery patients (pts) were sampled pre-tx and on days 1, 3, 5, 7, and 14 post-tx. Ten female trauma pts transfused with a total of 4 to 18 U of relatively fresh red blood cells were sampled up to 1.5 years post-tx. WBC subsets from frozen whole blood were isolated using CD4, CD8 (T cell), CD15 (myeloid), and CD19 (B cell) antibody-coated magnetic beads. Donor WBCs were counted by quantitative polymerase chain reaction (PCR) of male-specific sex determining region (SRY) sequences. PCR HLA typing and mixed leukocyte reaction (MLR) between recipient and donor WBCs were performed on two of the trauma tx recipients who had long-term chimerism of donor cells post-tx. In 6 of 8 female surgery pts, circulating CD4(+) male donor cells peaked at day 3 or 5 (0.01 to 1 cell/microL), followed by clearance by day 14. In 7 of 10 female trauma pts, we observed multilineage persistence of male donor WBCs (CD4, CD8, CD15, CD19) for 6 months to 1.5 years post-tx at concentrations of 10 to 100 cells/microL. In 2 trauma recipients studied, MLR showed no, or very low, response to WBC of the single donor implicated as the source of microchimerism by HLA typing. Establishment of long-term multilineage chimerism in trauma recipients is probably caused by engraftment of donor stem cells and mutual tolerance between recipient and donor leukocytes. A better understanding of factors determining clearance versus chimerism of transfused leukocytes is critical to prevention of alloimmunization and transfusion-induced graft-versus-host disease, and, potentially, to induction of tolerance for transplantation.
我们最近报道，在免疫功能正常的受者输血后3至5天，循环中的供体白细胞（WBC）出现短暂增加（《血液》85:1207，1995）。我们现在已经对其他输血人群中特定供体白细胞亚群的存活动力学进行了特征描述。对8名择期手术的女性患者在输血前以及输血后第1、3、5、7和14天进行采样。对10名总共输注了4至18单位相对新鲜红细胞的女性创伤患者，在输血后长达1.5年的时间内进行采样。使用包被有CD4、CD8（T细胞）、CD15（髓系细胞）和CD19（B细胞）抗体的磁珠从冷冻全血中分离白细胞亚群。通过对男性特异性性别决定区域（SRY）序列进行定量聚合酶链反应（PCR）来计数供体白细胞。对两名输血后有供体细胞长期嵌合现象的创伤患者进行了PCR HLA分型以及受者和供体白细胞之间的混合淋巴细胞反应（MLR）。在8名女性手术患者中的6名中，循环中的CD4(+)男性供体细胞在第3天或第5天达到峰值（0.01至1个细胞/微升），随后在第14天清除。在10名女性创伤患者中的7名中，我们观察到男性供体白细胞（CD4、CD8、CD15、CD19）多谱系持续存在于输血后6个月至1.5年，浓度为10至100个细胞/微升。在研究的2名创伤受者中，MLR显示对HLA分型确定为微嵌合体来源的单一供体的白细胞无反应或反应非常低。创伤受者中建立长期多谱系嵌合现象可能是由于供体干细胞的植入以及受者和供体白细胞之间的相互耐受。更好地理解决定输注白细胞清除与嵌合的因素对于预防同种免疫和输血诱导的移植物抗宿主病，以及潜在地诱导移植耐受性至关重要。
