van der Harst D, Brand A, van Luxemburg-Heys S A, Kooy-Winkelaar E M, van Rood J J
Department of Immunohematology and Blood Bank, University Medical Center, Leiden, The Netherlands.
Blood. 1989 Nov 15;74(7):2464-70.
Nine patients with leukemic B-lymphoproliferative diseases (B-LPD) were evaluated for development of in vitro recombinant interleukin-2 (rIL-2)-activated killer (LAK) cells. B-cell cultures were established from peripheral blood mononuclear cells (PBMNCs) containing 63% +/- 29% malignant cells. Short-term cultures were tested after 5-day activation with 500 U rIL-2/mL. Long-term cultures were maintained for 4 to 6 weeks by weekly addition of 500 U rIL-2 and autologous irradiated feeder cells. In the first week, the cells decreased considerably in the long-term cultures but thereafter cells proliferated (mainly T cells) on the average 300-fold (range 30- to 1,000-fold). In the short-term cultures, there was a 36% reduction of malignant B cells. In long-term cultures, B cells were reduced from 63% to 8%; three cultures still contained greater than 15% B cells. The CD16-positive cell percentage was comparable in both types of cultures and ranged from 2% to 17%. Effector cells lysing the natural killer (NK)-sensitive cell line K562 could be induced in all patients. Except in patients with chronic lymphocytic leukemia (CLL) and high malignant cell numbers, NK activity was already restored after 5 days. Optimal NK activity was obtained after 1.5 to 2.5 weeks. LAK cells killing NK-resistant lymphoma cell lines showed optimal activity after 2 to 3 weeks of culture. However, LAK cells killing greater than 10% of autologous malignant cells were obtained in only one third of the patients. The discrepancy between strong cytolytic activity against the NK-sensitive (K562) target cells obtained in all patients and the cytotoxic activity against NK-resistant cell lines contrasts with the poor development of LAK cells against autologous tumor cells. This discrepancy does not appear to be explained by soluble inhibitory factors released during the tumor cultures, as allogeneic LAK cells were not inhibited by supernatants from patients' cultures. Further investigations are warranted to reveal cell-mediated inhibition by tumor cells or suppressor cells.
对9例白血病性B淋巴细胞增殖性疾病（B-LPD）患者进行了体外重组白细胞介素-2（rIL-2）激活的杀伤细胞（LAK）培养发育评估。从含有63%±29%恶性细胞的外周血单个核细胞（PBMNC）建立B细胞培养物。用500 U rIL-2/mL激活5天后对短期培养物进行检测。通过每周添加500 U rIL-2和自体照射的饲养细胞将长期培养物维持4至6周。在第一周，长期培养物中的细胞显著减少，但此后细胞增殖（主要是T细胞），平均增殖300倍（范围为30至1000倍）。在短期培养物中，恶性B细胞减少了36%。在长期培养物中，B细胞从63%降至8%；三个培养物中仍含有超过15%的B细胞。两种培养物中CD16阳性细胞百分比相当，范围为2%至17%。所有患者均可诱导出裂解自然杀伤（NK）敏感细胞系K562的效应细胞。除慢性淋巴细胞白血病（CLL）患者和恶性细胞数量高的患者外，5天后NK活性已恢复。1.5至2.5周后获得最佳NK活性。培养2至3周后，杀伤NK抗性淋巴瘤细胞系的LAK细胞显示出最佳活性。然而，仅三分之一的患者获得了杀伤超过10%自体恶性细胞的LAK细胞。所有患者对NK敏感（K562）靶细胞具有强大的细胞溶解活性与对NK抗性细胞系的细胞毒性活性之间的差异，与LAK细胞对自体肿瘤细胞的发育不良形成对比。这种差异似乎不能用肿瘤培养过程中释放的可溶性抑制因子来解释，因为患者培养物的上清液并未抑制同种异体LAK细胞。有必要进行进一步研究以揭示肿瘤细胞或抑制细胞的细胞介导抑制作用。
