Kim Sung-Bae, Baskar Sivasubramanian, Kwak Larry W
Department of Medicine, Asan Medical Center, University of Ulsan, Seoul, South Korea.
Leuk Lymphoma. 2003 Jul;44(7):1201-8. doi: 10.1080/1042819031000076954.
In vitro priming of T cells with dendritic cells (DC) pulsed with clinically relevant, but weak antigens such as tumor idiotype (Id), is an attractive strategy to generate tumor-specific T lymphocytes. In order to enhance the specific antitumor effect of allogeneic stem cell grafts, we investigated whether induction of tumor specific T cells using autologous DC pulsed with patient's myeloma Id could be maintained and potentiated by in vitro priming. For induction of T cells, DC (5 x 10(5)/well) were cultured with autologous nonadherent cells (DoNA) (5 x 10(6)/well) and antigen (TT10 microg/ml, KLH 100 microg/ml and Id 100 microg/ml). The T cells were restimulated every 8-10 days with the corresponding antigen and autologons DC. After 2-4 cycles of in vitro priming, the T cells were compared with nonadherent cells obtained after 2h attachment on day 0 (DoNA) for antigen-specific cytokine production. In vitro primed T cells (2-4 cycles of stimulation with Ag and DC) showed significant antigen-specific cytokine responses (IFN-gamma, TNF-alpha, GM-CSF) to TT. Similarly, in vitro priming of T cells to Id-pulsed DC resulted in marked increases in cytokine production for both myeloma Id proteins tested. These data suggest that multiple in vitro immunization using DC could be beneficial in generating tumor specific T cells from normal donor PBMC, which may be used for adoptive immunotherapy (e.g. "tumor-specific" donor lymphocyte infusion) of B cell malignancies. In vitro immunization may also offer an alternative to immunization of healthy stem cell transplant donors with tumor antigen.
用与临床相关但较弱的抗原（如肿瘤独特型（Id））脉冲处理的树突状细胞（DC）在体外启动T细胞，是生成肿瘤特异性T淋巴细胞的一种有吸引力的策略。为了增强异基因干细胞移植的特异性抗肿瘤作用，我们研究了使用用患者骨髓瘤Id脉冲处理的自体DC诱导肿瘤特异性T细胞是否可以通过体外启动得以维持和增强。为了诱导T细胞，将DC（5×10⁵/孔）与自体非贴壁细胞（DoNA）（5×10⁶/孔）和抗原（TT 10μg/ml、KLH 100μg/ml和Id 100μg/ml）一起培养。T细胞每8 - 10天用相应抗原和自体DC重新刺激一次。经过2 - 4个体外启动周期后，将T细胞与第0天2小时贴壁后获得的非贴壁细胞（DoNA）比较抗原特异性细胞因子的产生。体外启动的T细胞（用抗原和DC刺激2 - 4个周期）对TT显示出显著的抗原特异性细胞因子反应（IFN -γ、TNF -α、GM - CSF）。同样，T细胞对Id脉冲处理的DC的体外启动导致所测试的两种骨髓瘤Id蛋白的细胞因子产生均显著增加。这些数据表明，使用DC进行多次体外免疫可能有利于从正常供体外周血单个核细胞中生成肿瘤特异性T细胞，这可用于B细胞恶性肿瘤的过继免疫治疗（如“肿瘤特异性”供体淋巴细胞输注）。体外免疫也可能为用肿瘤抗原免疫健康干细胞移植供体提供一种替代方法。
