Rohatiner A, Gelber R, Schlossman S F, Ritz J
Transplantation. 1986 Jul;42(1):73-80. doi: 10.1097/00007890-198607000-00016.
Graft-versus-host-disease (GVHD) remains the principal complication of allogeneic bone marrow transplantation. In animal models mature T lymphocytes have been shown to be responsible for GVHD and, therefore, in vitro treatment of donor bone marrow using monoclonal T cell specific antibodies and complement is currently being investigated as a strategy for the prevention of GVHD. In the present studies anti-T12 and anti-T11 monoclonal antibodies and rabbit complement were used to remove T lymphocytes from normal bone marrow. The efficacy of depletion was investigated by immunofluorescence assays and by in vitro culture of the residual cells using nonspecific mitogens or allogeneic B cells as the proliferative stimulus in the presence of lymphocyte-conditioned medium containing interleukin 2 (IL-2). Immunofluorescence analysis showed complete depletion of T12+ and T11+ cells after treatment with the respective antibodies and with the combination. Nevertheless, culture of treated bone marrow with phytohemagglutinin (PHA) or concanavalin A (Con A) and conditioned media containing IL-2 resulted in the proliferation of mature T cells (T3+, T4+ or T8+, T11+). Stimulation of treated marrow with allogeneic cells (Laz 388) resulted in the growth of a population with natural killer (NK) cell phenotype (T3-, T11+, NKH1+). The latter population was found to be strongly cytotoxic against K562 cells, a standard NK target. As expected, NK cells that are T11+ and T12- appeared to be more effected by in vitro treatment with anti-T11 than with anti-T12. A clonogenic assay was then used to quantitate the efficacy of target cell depletion in vitro. Three sequential incubations of bone marrow with either anti-T12 or anti-T11 plus complement resulted in depletion of 1-2 logs of clonogenic cells. Treatment with both antibodies concurrently resulted in elimination of 2-3 logs of clonogenic target cells. Although multiple treatments with both anti-T12 and anti-T11 were more effective than similar treatment with only one antibody, it remains to be established whether such combinations will be necessary in the clinical setting or whether more selective depletion of T cells without removal of NK cells might be optimal.
移植物抗宿主病（GVHD）仍然是同种异体骨髓移植的主要并发症。在动物模型中，已证明成熟T淋巴细胞是GVHD的病因，因此，目前正在研究使用单克隆T细胞特异性抗体和补体对供体骨髓进行体外处理，作为预防GVHD的一种策略。在本研究中，使用抗T12和抗T11单克隆抗体以及兔补体从正常骨髓中去除T淋巴细胞。通过免疫荧光测定以及在含有白细胞介素2（IL-2）的淋巴细胞条件培养基存在下，使用非特异性有丝分裂原或同种异体B细胞作为增殖刺激物对残留细胞进行体外培养，来研究去除效果。免疫荧光分析显示，在用各自抗体及联合使用两种抗体处理后，T12 +和T11 +细胞完全被去除。然而，用植物血凝素（PHA）或刀豆球蛋白A（Con A）以及含有IL-2的条件培养基培养经处理的骨髓，会导致成熟T细胞（T3 +、T4 +或T8 +、T11 +）增殖。用同种异体细胞（Laz 388）刺激经处理的骨髓，会导致具有自然杀伤（NK）细胞表型（T3 -、T11 +、NKH1 +）的细胞群体生长。发现后一细胞群体对标准NK靶细胞K562细胞具有强烈的细胞毒性。正如预期的那样，T11 +和T12 -的NK细胞似乎比抗T12抗体更易受抗T11抗体体外处理的影响。然后使用克隆形成试验来定量体外靶细胞去除的效果。用抗T12或抗T11加补体对骨髓进行三次连续孵育，导致克隆形成细胞减少1 - 2个对数。同时用两种抗体处理导致克隆形成靶细胞减少2 - 3个对数。尽管用抗T12和抗T11两种抗体进行多次处理比仅用一种抗体进行类似处理更有效，但在临床环境中是否需要这种联合处理，或者是否在不去除NK细胞的情况下更有选择地去除T细胞可能是最佳选择，仍有待确定。
