Barfield R C, Otto M, Houston J, Holladay M, Geiger T, Martin J, Leimig T, Gordon P, Chen X, Handgretinger R
Division of Stem Cell Transplantation, St Jude Children's Research Hospital, Memphis, TN 38105-2794, USA.
Cytotherapy. 2004;6(1):1-6. doi: 10.1080/14653240310004411.
The presence of T and B cells in allogeneic grafts contributes to GvHD and to EBV-associated lymphoproliferative disease (LPD). Depletion of T and B cells from the graft decreases the risk of these complications.
T and B cells were depleted from mobilized peripheral stem cells from volunteer donors (n=5) using anti-CD3 and anti-CD19 Abs conjugated to magnetic microbeads, and the CliniMACS device. The function of the stem cells after depletion was evaluated using colony assays and non-obese diabetic (NOD)/SCID repopulating experiments.
The mean mononuclear cell (MNC) count prior to T- and B-cell depletion was 2.19x10(10) (range 1.48-3.53). After depletion, the mean percentage of contaminating T cells was 0.02% (range 0.01-0.04%) with a mean log(10) depletion of 3.4 (range 3-3.8). The mean percentage of contaminating B cells was 0.1% (range 0.01-0.4%) with a mean log(10) depletion of 2.2 (range 1.4-3). The mean recovery of CD3- and CD19-negative MNCs after depletion was 70% (range 54-88%) and the mean recovery of CD34(+) stem cells was 69% (range 52-98%). The mean number of natural killer (NK) cells after T- and B-cell depletion was 5.2x10(8) (range 2-10x10(8)). In vitro colony assays and in vivo NOD/SCID repopulation assays showed no negative impact of this method on the function of the hematopoietic stem cells.
Our results show that the CliniMACS system can be used to efficiently deplete PBSC of T and B cells simultaneously, without adverse effect on the graft.
同种异体移植物中T细胞和B细胞的存在会导致移植物抗宿主病（GvHD）以及与EB病毒相关的淋巴增殖性疾病（LPD）。从移植物中去除T细胞和B细胞可降低这些并发症的风险。
使用与磁性微珠偶联的抗CD3和抗CD19抗体以及CliniMACS装置，从志愿供体（n = 5）动员的外周干细胞中去除T细胞和B细胞。使用集落测定法和非肥胖糖尿病（NOD）/重症联合免疫缺陷（SCID）再增殖实验评估去除后干细胞的功能。
T细胞和B细胞去除前单核细胞（MNC）的平均计数为2.19×10¹⁰（范围1.48 - 3.53）。去除后，污染T细胞的平均百分比为0.02%（范围0.01 - 0.04%），平均对数（10）去除率为3.4（范围3 - 3.8）。污染B细胞的平均百分比为0.1%（范围0.01 - 0.4%），平均对数（10）去除率为2.2（范围1.4 - 3）。去除后CD3和CD19阴性MNC的平均回收率为70%（范围54 - 88%），CD34⁺干细胞的平均回收率为69%（范围52 - 98%）。T细胞和B细胞去除后自然杀伤（NK）细胞的平均数量为5.2×10⁸（范围2 - 10×10⁸）。体外集落测定法和体内NOD/SCID再增殖测定法表明该方法对造血干细胞的功能没有负面影响。
我们的结果表明，CliniMACS系统可用于同时有效地去除外周血干细胞中的T细胞和B细胞，且对移植物无不良影响。
