Use of a SCID mouse/human lymphoma model to evaluate cytokine-induced killer cells with potent antitumor cell activity.
Use of a SCID mouse/human lymphoma model to evaluate cytokine-induced killer cells with potent antitumor cell activity.
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DOI：
10.1084/jem.174.1.139
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发表时间：
1991-07-01
期刊：
影响因子：
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通讯作者：
Weissman IL
中科院分区：
文献类型：
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作者：
Schmidt-Wolf IG;Negrin RS;Kiem HP;Blume KG;Weissman IL
C.B-17严重联合免疫缺陷（SCID）小鼠缺乏功能性B淋巴细胞和T淋巴细胞，允许异种移植，因此可以用于研究人类恶性肿瘤的生物学。将携带t（14;18）染色体易位的两种不同的人B细胞淋巴瘤细胞株SU-DHL-4和OCI-Ly8注射到C.B-17 SCID小鼠体内。静脉注射或腹腔注射的小鼠出现肿瘤并呈剂量依赖性死亡。肿瘤细胞存在于多种小鼠组织中，可以通过克隆性肿瘤实验、针对人B细胞抗原（CD19）的单克隆抗体（mAb）染色冷冻切片和聚合酶链反应技术来证明。一种使用细胞毒性效应细胞的方案被开发出来，并用于选择性地从骨髓中消耗肿瘤细胞。这些细胞是通过在干扰素γ (ifn - γ)、抗CD3单抗和白细胞介素2 （IL-2）存在下培养外周血单个核细胞发育而成的。如果在IL-2治疗前添加ifn - γ，则ifn - γ治疗的时机是关键和最佳的。经ifn - γ和IL-2刺激的细胞可通过靶向CD3的单克隆抗体进行扩增。这些细胞可以被IL-1进一步激活，但不能被肿瘤坏死因子α激活。采用该方案，通过克隆测定获得了3 log的肿瘤细胞杀伤。有趣的是，尽管这些细胞对淋巴瘤细胞具有很高的细胞毒活性，但对正常人类造血前体细胞（粒细胞/巨噬细胞集落形成单位）的亚群几乎没有毒性。这些细胞通过处理被人淋巴瘤细胞系SU-DHL-4污染的小鼠骨髓，并将这些细胞注射到SCID小鼠体内，以检测肿瘤的生长情况。SU-DHL-4细胞污染的骨髓经细胞因子诱导的杀伤细胞处理后，可提高小鼠的存活率。SCID小鼠为评估淋巴瘤治疗的新方法提供了一个有用的体内模型。细胞因子诱导的杀伤细胞在自体骨髓移植和过继免疫治疗中对骨髓清除有重要影响。
C.B-17 severe combined immune deficient (SCID) mice, which lack functional B and T lymphocytes, allow xenografts and, therefore, can be used to study the biology of human malignancies. Two different human B cell lymphoma cell lines, SU-DHL-4 and OCI-Ly8, which both harbor the t(14;18) chromosomal translocation, were injected into C.B-17 SCID mice. Mice injected intravenously or intraperitoneally developed tumors and died in a dose-dependent manner. The presence of tumor cells in various murine tissues could be demonstrated by a clonogenic tumor assay, staining of frozen sections with a monoclonal antibody (mAb) against a human B cell antigen (CD19), and with the polymerase chain reaction technique. A protocol using cytotoxic effector cells was developed and used to selectively deplete the tumor cells from bone marrow. These cells were developed by growing peripheral blood mononuclear cells in the presence of interferon gamma (IFN-gamma), anti- CD3 mAb, and interleukin 2 (IL-2). The timing of IFN-gamma treatment was critical and optimal if IFN-gamma was added before IL-2 treatment. The cells that were stimulated by IFN-gamma, followed by IL-2, could be expanded by treatment with a mAb directed against CD3. These cells could be further activated by IL-1, but not by tumor necrosis factor alpha. With this protocol, a tumor cell kill of 3 logs was obtained as measured by a clonogenic assay. Interestingly, despite their high cytotoxic activity against lymphoma cells, these cells had little toxicity against a subset of normal human hematopoietic precursor cells (granulocyte/macrophage colony-forming units). These cells were further tested by treating murine bone marrow contaminated with the human lymphoma cell line SU-DHL-4, and injecting these cells into SCID mice to assay for tumor growth in vivo. The animals injected with bone marrow contaminated with SU-DHL-4 cells had enhanced survival if the bone marrow was treated with the cytokine-induced killer cells before infusion. The SCID mouse provides a useful in vivo model for evaluation of new therapeutic approaches for lymphoma treatment. The cytokine- induced killer cells generated as described here could have an important impact on bone marrow purging for autologous bone marrow transplantation as well as for adoptive immunotherapy.