Rocha M, Umansky V, Lee K H, Hacker H J, Benner A, Schirrmacher V
Deutsches Krebsforschungszentrum Heidelberg, Abteilung Zelluläre Immunologie, FSP Tumorimmunologie, Germany.
Blood. 1997 Mar 15;89(6):2189-202.
Graft-versus-leukemia (GVL) and Graft-versus-host (GVH) reactions were compared after systemic transfer of allogeneic antitumor immune T lymphocytes from B10.D2 (H-2d; Mls(b)) into DBA/2 (H-2d; Mis(a)) mice. Before immune cell transfer, recipient DBA/2 mice were sublethally irradiated with 5 Gy to prevent host-versus-graft reactivity. Recipients were either bearing syngeneic metastatic ESb lymphomas (GVL system) or were normal, non-tumor-bearing mice (GVH system). We previously reported that this adoptive immunotherapy protocol (ADI) had pronounced GVL activity and led to immune rejection of even advanced metastasized cancer. In this study, monoclonal antibodies were used for immunohistochemical analysis of native frozen tissue sections from either spleen or liver to distinguish donor from host cells, to differentiate between CD4 and CD8 T lymphocytes, and to stain sialoadhesin-positive macrophages at different time points after cell transfer. The kinetics of donor cell infiltration in spleen and liver differed in that the lymphoid organ was infiltrated earlier (days 1 to 5 after transfer) than the nonlymphoid organ (days 5 to 20). After reaching a peak, donor cell infiltration decreased gradually and was not detectable in the spleen after day 20 and in the liver after day 30. The organ-infiltrating donor immune cells were mostly T lymphocytes and stained positive for CD4 or CD8 T-cell markers. A remarkable GVL-associated observation was made with regard to a subset of macrophages bearing the adhesion molecule sialoadhesin (SER+ macrophages). In the livers of tumor-bearing mice, their numbers increased between days 1 and 12 after ADI by a factor greater than 30. Double-staining for donor cell marker and SER showed that the sialoadhesin-expressing macrophages were of host origin. The SER+ host macrophages from GVL livers were isolated by enzyme perfusion and rosetting 12 days after ADI, when they reached peak values of about 60 cells per liver lobule, and were tested, without further antigen addition, for their capacity to stimulate an antitumor CD8 T-cell response. The results of this immunologic analysis suggest that these cells in the liver function as scavengers of the destroyed metastases and as antigen-processing and -presenting cells for antitumor immune T cells.
将来自B10.D2（H-2d；Mls(b)）的同种异体抗肿瘤免疫T淋巴细胞全身转移至DBA/2（H-2d；Mls(a)）小鼠后，对移植物抗白血病（GVL）反应和移植物抗宿主（GVH）反应进行了比较。在免疫细胞转移前，对受体DBA/2小鼠进行5 Gy的亚致死剂量照射，以防止宿主抗移植物反应。受体小鼠要么携带同基因转移性ESb淋巴瘤（GVL系统），要么是正常的、无肿瘤的小鼠（GVH系统）。我们之前报道过，这种过继性免疫治疗方案（ADI）具有显著的GVL活性，甚至能导致晚期转移性癌症的免疫排斥。在本研究中，使用单克隆抗体对脾脏或肝脏的天然冷冻组织切片进行免疫组织化学分析，以区分供体细胞和宿主细胞，区分CD4和CD8 T淋巴细胞，并在细胞转移后的不同时间点对唾液酸粘附素阳性巨噬细胞进行染色。供体细胞在脾脏和肝脏中的浸润动力学有所不同，即淋巴器官比非淋巴器官浸润得更早（转移后1至5天）（非淋巴器官浸润时间为5至20天）。达到峰值后，供体细胞浸润逐渐减少，在第20天后脾脏中以及第30天后肝脏中均检测不到。浸润器官的供体免疫细胞主要是T淋巴细胞，对CD4或CD8 T细胞标志物呈阳性染色。关于携带粘附分子唾液酸粘附素的巨噬细胞亚群（SER+巨噬细胞）有一个与GVL相关的显著观察结果。在荷瘤小鼠的肝脏中，它们的数量在ADI后第1天至第12天之间增加了30倍以上。对供体细胞标志物和SER进行双重染色显示，表达唾液酸粘附素的巨噬细胞来自宿主。在ADI后12天，当GVL肝脏中SER+宿主巨噬细胞达到每个肝小叶约60个细胞的峰值时，通过酶灌注和花环沉降法分离出这些细胞，在不添加进一步抗原的情况下，检测它们刺激抗肿瘤CD8 T细胞反应的能力。这项免疫分析结果表明，肝脏中的这些细胞起到清除被破坏转移灶的作用，并作为抗肿瘤免疫T细胞的抗原处理和呈递细胞。
