自体及异体CD3单克隆抗体激活杀伤细胞对正常造血细胞的影响

Effect of Autologous or Allogeneic Anti-CD3 Monoclonal Antibody Activated Killer Cells on Normal Hematopoietic Cells

为探索自体及异体CD3单克隆抗体激活杀伤 (CD3AK)细胞对正常造血细胞的影响 ,以固化抗CD3单克隆抗体联合小剂量IL 2诱导CD3AK细胞。采用流式细胞术 (FCM )检测CD3AK细胞的表型变化 ,并检测正常骨髓经与自体或异体CD3AK细胞培养后CD34+ 细胞的比例变化 ,用集落培养检测正常骨髓经与自体或异体CD3AK细胞共同培养后CFU GM的计数变化。结果显示 :3- 5 μg ml固化抗CD3单克隆抗体联合 10 0U mlIL 2可有效地激活CD3AK细胞 ;正常骨髓与异体CD3AK细胞培养 6小时后CD34+ 细胞的比例下降 32 .37%,正常骨髓与自体CD3AK细胞共同培养 6小时后CD34+ 细胞比例升高 5 .4 9%;CFU GM集落培养显示正常骨髓与异体CD3AK细胞共同培养 6小时后CFU GM的计数下降 2 0 .4 4 %,而正常骨髓与自体CD3AK细胞共同培养后CFU GM的数量下降 3.39%。上述结果表明 ,异体CD3AK细胞与正常骨髓造血细胞短期共同培养可产生一定的抑制作用 ,自体CD3AK细胞与骨髓造血细胞短期共同培养无明显影响。

In order to investigate the effect of autologous and allogeneic anti-CD3 McAb activated killer cells(CD3AK) on normal hematopoietic cells, the immobilized anti-CD3 McAb and low concentration IL-2 were used to activate CD3AK. Flow cytometry was used to assay the phenotype change of CD3AK to analyze the proportional change of CD34 + cells in normal bone marrow mononuclear cells(BMMNC) cocultured with autologous or allogeneic CD3AK. The effect of CD3AK on normal hematopoietic progenitor cells was also assayed by methylcellulose clonogenic culture of CFU-GM. It was found that 3-5 μg/ml immobilized anti-CD3 McAb and 100 U/ml IL-2 could activate CD3AK effectively. There were 99.51% CD3 + cells in CD3AK groups. When BMMNCs from healthy volunteers were cocultured with allogeneic CD3AK for six hours, the percentage of CD34 + cells was decreased 32.37%. CD3AK had no significant influence on autologous BMMNC. Allogeneic and autologous CD3AK were cultured with BMMNC from healthy volunteers for six hours, and then CFU-GM was evaluated. Allogeneic CD3AK inhibited 20.44% CFU-GM formation, but autologous CD3AK had no inhibition on CFU-GM. It is concluded that CD3AK has no inhibition to autologous normal hematopoietic progenitor cells after cocultured with them from these results, while allogeneic CD3AK inhibits the normal hematopoietic progenitor cells significantly.