Sondel P M, Hank J A, Wendel T, Flynn B, Bozdech M J
J Clin Invest. 1983 Jun;71(6):1779-86. doi: 10.1172/jci110933.
Lymphocytes from a healthy HLA-identical bone marrow transplant donor were tested for their ability to destroy her brother's acute myelogenous leukemia blasts in vitro. Primary mixed lymphocyte culture (MLC) and cell-mediated lysis (CML) responses between the patient's remission (pretransplant) and donor's lymphocytes were negative. Stimulation of donor lymphocytes for 7 d in vitro with irradiated leukemia cells, leukemia cells plus allogeneic irradiated lymphocytes, or a pool of irradiated lymphocytes from 10 donors, did not activate any cytotoxic cells able to destroy the HLA identical leukemic blasts. Further culturing for 7 additional d in T cell growth factor (TCGF) generated lymphocytes that induced effective cytotoxicity against the leukemic blasts, but not against autologous lymphocytes. Effective killing against the leukemia was observed only in cultures initially stimulated with the irradiated leukemia cells. These cytotoxic cells were maintained in TCGF and mediated persistent killing against the leukemic target cells. They were also able to destroy lymphocytes from the patient's mother and father, but not from an unrelated cell donor. This suggested specific recognition of non-HLA antigens inherited by the patient, that were foreign to the HLA identical bone marrow donor. These lymphocytes were cloned by a limiting dilution technique and one clone maintained cytotoxicity to the AML blasts and the father's lymphocytes, but not lymphocytes from the mother or an HLA-identical donor. This cytotoxicity was inhibited by a monoclonal anti-HLA antibody. Thus, in vitro sensitization of this sibling's lymphocytes with AML blasts followed by TCGF expansion, and cloning, enabled the detection of HLA-restricted cytotoxic cells that recognize minor locus histocompatibility antigens. This immune recognition may be relevant to the "graft vs. leukemia" effect that has been observed in leukemic animals and patients following histocompatible hematopoietic transplants.
对来自一名健康的 HLA 相同的骨髓移植供体的淋巴细胞进行体外测试，以检测其破坏她哥哥急性髓性白血病原始细胞的能力。患者缓解期（移植前）与供体淋巴细胞之间的初次混合淋巴细胞培养（MLC）和细胞介导的裂解（CML）反应均为阴性。用辐照的白血病细胞、白血病细胞加异体辐照淋巴细胞或来自 10 名供体的辐照淋巴细胞池对供体淋巴细胞进行体外刺激 7 天，未激活任何能够破坏 HLA 相同白血病原始细胞的细胞毒性细胞。在 T 细胞生长因子（TCGF）中再培养 7 天产生的淋巴细胞可诱导对白血病原始细胞有效细胞毒性，但对自体淋巴细胞无效。仅在最初用辐照白血病细胞刺激的培养物中观察到对白血病的有效杀伤。这些细胞毒性细胞在 TCGF 中得以维持，并介导对白血病靶细胞的持续杀伤。它们还能够破坏患者父母的淋巴细胞，但不能破坏无关细胞供体的淋巴细胞。这表明对患者遗传的非 HLA 抗原具有特异性识别，这些抗原对于 HLA 相同的骨髓供体来说是外来的。这些淋巴细胞通过有限稀释技术进行克隆，一个克隆对 AML 原始细胞和父亲的淋巴细胞保持细胞毒性，但对母亲或 HLA 相同供体的淋巴细胞无细胞毒性。这种细胞毒性被单克隆抗 HLA 抗体抑制。因此，用 AML 原始细胞对该同胞的淋巴细胞进行体外致敏，随后进行 TCGF 扩增和克隆，能够检测到识别次要组织相容性抗原的 HLA 限制性细胞毒性细胞。这种免疫识别可能与在组织相容性造血移植后的白血病动物和患者中观察到的“移植物抗白血病”效应相关。
