冻融抗原负载树突状细胞诱导细胞毒性T淋巴细胞特异性杀伤急性白血病细胞的实验研究

我们应用基因重组人白介素 4 (rhIL 4 ) ,基因重组人粒 /单细胞集落刺激因子 (rhGM CSF)联合培养体系自健康志愿者骨髓单个核细胞诱导产生DC ,使其负载急性髓系白血病 (AML)细胞冻融抗原 ,体外诱导细胞毒性T细胞 (CTL) ,观察负载AML细胞冻融抗原后DC的状态对所激发的T淋巴细胞表型及功能的影响及CTL对AML细胞特异性杀伤活性。我们发现负载AML冻融抗原后DC的CD1a、CD83、CD86、CD11C、HLA DR表达率为较培养前明显增高 (P <0 .0 1) ,且负载AML冻融抗原的DC诱导的CTL中CD3 CD8 T细胞比例 ,较诱导前明显增高 (P <0 .0 1) ,而负载AML细胞冻融抗原的DC诱导CTL对AML细胞有较强的杀伤作用 ,明显强于加或不加IL 2培养的T细胞对照组 (P <0 .0 1) ,而对K5 6 2细胞无明显的杀伤活性。通过以上实验我们认为经rhGM CSF ,rhIL 4培养产生的DC为CD14CD1a DC ,能诱导CTL对AML细胞产生明显的特异性杀伤作用 ,另外 ,负载AML细胞冻融抗原DC诱导的CTL中CD3 CD8 T细胞比例明显增高 ,提示CD8 T细胞在抗肿瘤免疫中具有极其重要的作用。

Investigation on Specific Killing Acute Leukemia Cell Reaction of the Cytotoxic T Lymphocyte Induced by Dendritic Cell Pulsed with Frozen-thawed Antigen

We investigated the effects of dendritic cell(DC) pulsed with acute leukemia cell frozen-thawed antigen on inducing the cytotoxic T lymphocyt e (CTL) to get specific anti-tumor activity in vitro. DC was generated from healthy human bone marrow mononuclear cell (BMMC) in the presence of granulocyt e/mac rophage-colony stimulating factor(GM-CSF),interleukin-4(IL-4) in vitro. DC pulsed with acute leukemia ce ll frozen-thawed antigen was co-cultured to induce T cell into specific CTL. T hen we observed the effects of CTL induced by DC pulsed with acute leukemia cell frozen- t hawed antigen killing acute leukemia cell specially and the influence of dendrit ic cell affecting the function and CD expression on CTL.The levels of CD1a, CD86 , HLA-DR ex pression on DC pulsed with acute leukemia cell frozen-thawed antigen were obvio usly higher than those before culture (P<0 01). There were more CD3 CD8 T ce lls in the CTL induced by DC pulsed with acute leukemia cell frozen-thawed anti gen, compared with those in the T cell uncultured group (P<0.01).The CTL ind uced by DC pulsed with acute leukemia cell frozen-thawed antigen significantly had higher activity in killing acute leukemia cell than in killing k562 cell ( P<0.01),and the CTL induced by DC pulsed with acut e leukemia cell frozen-thawed antigen was also more effective for killing acute leukemia cell as compared with the CTL induced by DC simply,T cell co-culture d with IL-2 and T cell simply(P<0.01).The DC generated from human bone marr ow mononuclear cell(BMM C) in the presence of granulocyte/macrophage-colony stimulating factor(GM-CSF) ,i nterleukin-4(IL-4) was CD14- CD1a CD83 DC,and it could also induce the c ytotoxic T lymphocyte(CTL) to get specific anti-tumor activity in vitro.Otherwise,th e inc reasing of CD3 CD8 T cells in the CTL induced by DC pulsed with acute leuk e mia cell frozen-thawed antigen implied the main role of the CD3 CD8 T cell s in the anti-tumor immunity.