Gambacorti-Passerini C, Rivoltini L, Fizzotti M, Rodolfo M, Sensi M L, Castelli C, Orazi A, Polli N, Bregni M, Siena S
Division of Experimental Oncology, Istituto Nazionale Tumori-Milan, Italy.
Br J Haematol. 1991 Jun;78(2):197-205. doi: 10.1111/j.1365-2141.1991.tb04416.x.
The ability of recombinant interleukin 2 (rIL2) activated lymphocytes (LAK) to purge BM samples contaminated by tumour cells was evaluated. Human BM mononuclear cells were contaminated with 10% of the lymphoma line CA46 and then cultured in liquid medium containing 1000 U/ml of rIL2 and/or LAK autologous to the used BM. At the end of coculture the growth of residual tumour cells and of CFU-GM were evaluated by clonogenic assay. No tumour cell growth was observed in 5/5 independent experiments after 18 h of coculture with LAK. No significant inhibition of CFU-GM growth was also noted. Subsequently, the effect of LAK on BM obtained from four leukaemic patients and contaminated with 20-50% of their own AML and ALL cells was studied using MAb as a tool for identifying leukaemic cells. LAK eliminated 24-78% of contaminating cryopreserved uncultured autologous leukaemic cells. In five cases the BM was contaminated by a low (2%) amount of ALL cells. In these patients the monoclonal heavy chain rearrangement typical of ALL was no longer visible after coculture with LAK. Evidence for selective tumour cytotoxicity by LAK was confirmed by using autologous BM cells as hot and cold targets in a 51Cr release assay. Finally, successful haematologic reconstitution of lethally irradiated BALB/c mice was obtained using syngeneic BM cocultured with LAK. These results support the investigational use of rIL2 and LAK in the treatment of human leukaemia.
评估了重组白细胞介素2（rIL2）激活的淋巴细胞（LAK）清除被肿瘤细胞污染的骨髓样本的能力。人骨髓单个核细胞被10%的淋巴瘤细胞系CA46污染，然后在含有1000 U/ml rIL2和/或与所用骨髓自体的LAK的液体培养基中培养。共培养结束时，通过克隆形成试验评估残留肿瘤细胞和CFU-GM的生长情况。与LAK共培养18小时后，在5个独立实验中均未观察到肿瘤细胞生长。也未发现CFU-GM生长受到显著抑制。随后，以单克隆抗体（MAb）作为识别白血病细胞的工具，研究了LAK对从4例白血病患者获得的、被20%-50%自身急性髓系白血病（AML）和急性淋巴细胞白血病（ALL）细胞污染的骨髓的作用。LAK清除了24%-78%的冷冻保存的未培养自体白血病污染细胞。在5例中，骨髓被少量（2%）ALL细胞污染。在这些患者中，与LAK共培养后，ALL典型的单克隆重链重排不再可见。通过在51Cr释放试验中使用自体骨髓细胞作为热靶和冷靶，证实了LAK具有选择性肿瘤细胞毒性。最后，使用与LAK共培养的同基因骨髓，成功实现了对致死性照射的BALB/c小鼠的血液学重建。这些结果支持rIL2和LAK在人类白血病治疗中的研究应用。
