T-cell immunotherapy with a chimeric receptor against CD38 is effective in eradicating chemotherapy-resistant B-cell lymphoma cells overexpressing survivin induced by BMI-1.
T-cell immunotherapy with a chimeric receptor against CD38 is effective in eradicating chemotherapy-resistant B-cell lymphoma cells overexpressing survivin induced by BMI-1.
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DOI：
10.1038/bcj.2012.21
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发表时间：
2012-06
影响因子：
12.8
通讯作者：
Takihara, Y.
中科院分区：
文献类型：
--
作者：
Bhattacharyya, J.;Mihara, K.;Kitanaka, A.;Yanagihara, K.;Kubo, T.;Takei, Y.;Kimura, A.;Takihara, Y.
BMI-1（B淋巴瘤Mo-MLV插入区1同源物）是多梳组基因（PcG）的一个成员，其表达与恶性肿瘤患者的不良预后和治疗失败密切相关，所述恶性肿瘤例如骨髓增生异常综合征、慢性髓性白血病、急性髓性白血病和淋巴瘤。1-3最近，我们发现BMI-1通过诱导生存素的表达使B细胞淋巴瘤细胞对几种抗癌药物无效。临床上迫切需要找到治疗方法来治疗过度表达BMI-1和生存素的淋巴瘤细胞患者。尽管在前利妥昔单抗时代（利妥昔单抗，IDEC Pharmaceuticals，San Diego，CA，USA），弥漫性大B细胞淋巴瘤（DLBCL）患者的长期缓解率为50- 60%，但利妥昔单抗的加入已导致生存率的巨大改善。由于利妥昔单抗比具有生发中心B细胞型DLBCL的患者更显著地改善了具有不良预后的非生发中心B细胞型DLBCL患者的总体存活、无事件存活和无进展存活，5-9使用抗体如利妥昔单抗的免疫疗法可以通过克服包括Bcl-2、Bcl-6和Bcl-xL的耐药基因而起作用。我们先前开发了具有针对CD 38的嵌合抗原受体（CAR）的T细胞，并报道这些CD 38特异性T细胞在体外和体内有效地消除了B细胞淋巴瘤细胞。11，12这是因为CD 38在B细胞淋巴瘤细胞中广泛且高度表达（40-50%的B细胞型患者），13特别是在艾滋病相关淋巴瘤细胞14和具有侵袭性的携带t（14; 18）的DLBCL细胞中（100%的这些患者）。因此，我们研究了携带抗CD 38-CAR的T细胞是否对表达BMI-1和存活素的B细胞淋巴瘤细胞产生细胞毒性。在这里，我们报告说，CD 38特异性T细胞有效地消除化疗耐药B淋巴瘤细胞过度表达BMI-1和生存素，并提出T细胞免疫疗法与CAR可能是有用的治疗难治性B细胞淋巴瘤。两个淋巴瘤细胞系（HT和RL），获自美国典型培养物保藏中心（ATCC）（Manassas，VA，USA）在补充有10%胎牛血清（FCS）的RPMI-1640培养基中于37 ℃培养。用含有MSCV-BMI-1-Flag-IRES-GFP或单独的MSCV-IRES-GFP的水泡性口炎病毒G糖蛋白（VSVG）假型逆转录病毒转导细胞，并通过FACS Aria（BD，圣何塞，CA，USA）分选GFP阳性细胞。从淋巴瘤患者的淋巴结、胸腔积液和脾脏以及健康供体的外周血细胞获得原代细胞，并进行Ficoll密度离心。获得所有患者和供体的知情同意书。经广岛大学机构审查委员会批准，对B细胞淋巴瘤患者和供体进行检查。先前制备了由GFP、CD 8a、4-1BB、CD 3 z的跨膜结构域和抗CD 38 scFv组成的逆转录病毒载体构建体。11
The expression of BMI-1 (B lymphoma Mo-MLV insertion region 1 homolog), a member of the polycomb-group genes (PcG), is well correlated with a poor prognosis and treatment failure among patients with malignancies such as myelodysplastic syndrome, chronic myeloid leukemia, acute myeloid leukemia and lymphoma. 1–3 Recently, we found that BMI-1 renders B-cell lymphoma cells refractory to several anti-cancer drugs by inducing the expression of survivin. 4 There is an urgent clinical need to find therapeutics to treat patients with lymphoma cells overexpressing BMI-1 and survivin. Although in the pre-rituximab era (Rituximab, IDEC Pharmaceuticals, San Diego, CA, USA), the long-term remission rate for patients with diffuse large B-cell lymphoma (DLBCL) was 50–60%, the addition of rituximab has led to an enormous improvement in survival. As rituximab has more significantly improved the overall survival, event-free survival and progression-free survival of patients with non-germinal center B cell-type DLBCL, which have a poor prognosis, than those with germinal center B cell-type DLBCL, 5–9 immunotherapy with an antibody such as rituximab may function by overcoming drugresistant genes including Bcl-2, Bcl-6 and Bcl-xL. 10 We previously developed T cells with a chimeric antigen receptor (CAR) against CD38 and reported that these CD38-specific T cells effectively eliminated B-cell lymphoma cells in vitro and in vivo. 11, 12 This is because CD38 is widely and highly expressed in B-cell lymphoma cells (40–50% of patients with the B-cell typed), 13 especially in AIDS-associated lymphoma cells14 and DLBCL cells bearing t (14; 18) with aggressiveness (100% of these patients). 15 We, thus investigated whether T cells bearing an anti-CD38-CAR exerted cytotoxicity against B-cell lymphoma cells expressing BMI-1 and survivin. Here we report that, the CD38-specific T cells efficiently eliminated chemotherapy-resistant B-lymphoma cells overexpressing BMI-1 and survivin, and propose that T-cell immunotherapy with CAR may be useful for treating refractory B-cell lymphoma.Two lymphoma cell lines (HT and RL) obtained from American Type Culture Collection (ATCC)(Manassas, VA, USA) were cultured in RPMI-1640 medium supplemented with 10% fetal calf serum (FCS) at 37 1C. The cells were transduced with a vesicular stomatitis virus G glycoprotein (VSVG)-pseudotyped retrovirus containing MSCV-BMI-1-Flag-IRES-GFP or MSCV-IRES-GFP alone and the GFP-positive cells were sorted by FACS Aria (BD, San Jose, CA, USA). Primary cells were obtained from the lymph node, pleural effusion, and spleen of patients with lymphoma, and peripheral blood cells from healthy donors, and subjected to Ficoll-density centrifugation. Informed consent was obtained from all of the patients and donors. Patients with B-cell lymphoma and donors were examined as approved by the institutional review board at Hiroshima University. The retroviral vector construct consisting of GFP, the transmembrane domain of CD8a, 4-1BB, CD3z and anti-CD38 scFv was made previously. 11
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影响因子：
45.3
作者：
Fu, Kai;Weisenburger, Dennis D.;Vose, Julie M.
通讯作者：
Vose, Julie M.
带有嵌合抗原受体的T细胞具有有效的抗肿瘤作用，可以在晚期白血病患者中建立记忆。
影响因子：
17.1
作者：
Kalos M;Levine BL;Porter DL;Katz S;Grupp SA;Bagg A;June CH
通讯作者：
June CH
影响因子：
10.1
作者：
Le Gouill, Steven;Talmant, Pascaline;Avet-Loiseau, Herve
通讯作者：
Avet-Loiseau, Herve
影响因子：
6.5
作者：
Mihara, Keichiro;Yanagihara, Kazuyoshi;Kimura, Akiro
通讯作者：
Kimura, Akiro
DOI：
10.1056/nejmoa1103849
发表时间：
2011-08-25
期刊：
The New England journal of medicine
影响因子：
--
作者：
Porter DL;Levine BL;Kalos M;Bagg A;June CH
通讯作者：
June CH