Schmidt-Wolf I G, Negrin R S, Kiem H P, Blume K G, Weissman I L
Department of Medicine, Stanford University Medical Center, California 94305.
J Exp Med. 1991 Jul 1;174(1):139-49. doi: 10.1084/jem.174.1.139.
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.
C.B-17重度联合免疫缺陷（SCID）小鼠缺乏功能性B淋巴细胞和T淋巴细胞，能够接受异种移植，因此可用于研究人类恶性肿瘤的生物学特性。将两种携带t(14;18)染色体易位的不同人类B细胞淋巴瘤细胞系SU-DHL-4和OCI-Ly8注射到C.B-17 SCID小鼠体内。静脉或腹腔注射的小鼠会以剂量依赖的方式发生肿瘤并死亡。通过克隆形成肿瘤试验、用抗人B细胞抗原（CD19）的单克隆抗体（mAb）对冰冻切片进行染色以及聚合酶链反应技术，可以证明各种鼠组织中存在肿瘤细胞。开发了一种使用细胞毒性效应细胞的方案，并用于从骨髓中选择性清除肿瘤细胞。这些细胞是通过在γ干扰素（IFN-γ）、抗CD3 mAb和白细胞介素2（IL-2）存在的情况下培养外周血单个核细胞而产生的。IFN-γ治疗的时机至关重要，如果在IL-2治疗之前添加IFN-γ则效果最佳。先由IFN-γ刺激，然后用IL-2刺激的细胞，可以通过用抗CD3的mAb处理而扩增。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。按照此方案，通过克隆形成试验测得肿瘤细胞杀伤率为3个对数。有趣的是，尽管这些细胞对淋巴瘤细胞具有高细胞毒性活性，但对一部分正常人造血前体细胞（粒细胞/巨噬细胞集落形成单位）几乎没有毒性。通过处理被人类淋巴瘤细胞系SU-DHL-4污染的鼠骨髓，并将这些细胞注射到SCID小鼠体内以检测体内肿瘤生长，对这些细胞进行了进一步测试。如果在输注前用细胞因子诱导的杀伤细胞处理骨髓，那么注射了被SU-DHL-4细胞污染的骨髓的动物存活率会提高。SCID小鼠为评估淋巴瘤治疗的新治疗方法提供了一个有用的体内模型。本文所述产生的细胞因子诱导的杀伤细胞可能对自体骨髓移植的骨髓净化以及过继性免疫治疗产生重要影响。
