通过抗CD3×抗CD19双特异性抗体联合阿糖胞苷重定向CD4+和CD8+ T淋巴细胞以及有效裂解患者来源的B-ALL细胞。
Redirection of CD4+ and CD8+ T lymphocytes via an anti-CD3 × anti-CD19 bi-specific antibody combined with cytosine arabinoside and the efficient lysis of patient-derived B-ALL cells.
作者信息
Fan Dongmei, Li Wei, Yang Yuqi, Zhang Xiaolong, Zhang Qing, Yan Yan, Yang Ming, Wang Jianxiang, Xiong Dongsheng
机构信息
State Key Laboratory of Experimental Hematology, Institute of Hematology and Hospital of Blood Diseases, Chinese Academy of Medical Sciences and Peking Union Medical College, Tianjin, 300020, People's Republic of China.
Department of Maxillofacial and E.N.T. Oncology, Tianjin Medical University Cancer Institute and Hospital, National Clinical Research Center of Cancer, Key Laboratory of Cancer Prevention and Therapy, Tianjin, 300060, People's Republic of China.
出版信息
J Hematol Oncol. 2015 Oct 6;8:108. doi: 10.1186/s13045-015-0205-6.
BACKGROUND
B-acute lymphoblastic leukemia (B-ALL) is derived from B cell progenitors. Recently, the development of appropriate combinations of chemotherapy and immunotherapy represents a promising approach for eliminating cancer. We previously constructed an anti-CD3 × anti-CD19 bi-specific antibody in a diabody configuration and its disulfide-stabilized format (ds-diabody). The combination of the diabody or ds-diabody and Ara-C was highly effective in enhancing the cytotoxicity of T cells against the CD19+ human leukemia cell-line, Nalm-6, both in vitro and in vivo. This study verified whether B-ALL patient-derived cells were sensitive to the diabody or ds-diabody and low-dosage Ara-C combination.
METHODS
This study aimed to detect the B7 family members B7.1 (CD80) and B7.2 (CD86) that were expressed in B-ALL patient-derived cells pre-treated by Ara-C (0.25 μM) and to determine the targeted killing ability of T cell subtypes induced by the diabody or ds-diabody combination with Ara-C both in vitro and in vivo. We also determined the levels of the cytokines that were released by activated CD4+ or CD8+ T cells during therapy.
RESULT
Low-dose Ara-C enhanced CD80 and CD86 expression in nearly 50 % of specimens of B-ALL patient-derived cells. A combination of diabody or ds-diabody and Ara-C enhanced T cell against B-ALL cells in vitro and in vivo. Both CD8+ and CD4+ T cells were potently activated. Expression of CD25 and CD69 was augmented equally by CD4+ or CD8+ T cells. However, CD8+ T cells made the major contribution by redirecting target cell lysis in a granzyme B and perforin-dependent mechanism. CD4+ T cells played an important immunomodulatory role by secreting IL2. Consequently, IL3, IL6, TNFα, and IFNγ were also released by CD4+ or CD8+ T cells following diabody-mediated T cell activation.
CONCLUSION
T cell therapy induced by diabody or ds-diabody combined with low dose of Ara-C was effective against cancer cell-lines and in clinical trials. In vivo, the ds-diabody was more efficient than its parent diabody due to its enhanced stability.
背景
B 细胞急性淋巴细胞白血病（B-ALL）起源于 B 细胞祖细胞。最近，化疗与免疫疗法的适当联合应用为癌症治疗带来了新的希望。我们之前构建了一种双体结构的抗 CD3×抗 CD19 双特异性抗体及其二硫键稳定形式（ds-双体）。双体或 ds-双体与阿糖胞苷联合使用，在体外和体内均能显著增强 T 细胞对 CD19+人白血病细胞系 Nalm-6 的细胞毒性。本研究旨在验证 B-ALL 患者来源的细胞对双体或 ds-双体与低剂量阿糖胞苷联合治疗是否敏感。
方法
本研究旨在检测经阿糖胞苷（0.25 μM）预处理的 B-ALL 患者来源细胞中表达的 B7 家族成员 B7.1（CD80）和 B7.2（CD86），并确定双体或 ds-双体与阿糖胞苷联合诱导的 T 细胞亚群在体外和体内的靶向杀伤能力。我们还测定了治疗过程中活化的 CD4+或 CD8+ T 细胞释放的细胞因子水平。
结果
低剂量阿糖胞苷可使近 50%的 B-ALL 患者来源细胞标本中 CD80 和 CD86 的表达增强。双体或 ds-双体与阿糖胞苷联合使用可在体外和体内增强 T 细胞对 B-ALL 细胞的杀伤作用。CD8+和 CD4+ T 细胞均被有效激活。CD4+或 CD8+ T 细胞均可同等程度地增强 CD25 和 CD69 的表达。然而，CD8+ T 细胞通过颗粒酶 B 和穿孔素依赖的机制重定向靶细胞裂解发挥主要作用。CD4+ T 细胞通过分泌 IL2 发挥重要的免疫调节作用。因此，双体介导的 T 细胞活化后，CD4+或 CD8+ T 细胞也会释放 IL3、IL6、TNFα 和 IFNγ。
结论
双体或 ds-双体联合低剂量阿糖胞苷诱导的 T 细胞疗法对癌细胞系有效，并已进入临床试验阶段。在体内，ds-双体由于稳定性增强，比其亲本双体更有效。
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