Attisano C, Bianchi A, Montacchini L, Carlesso N, Peola S, Bruno B, Roux V, Ferrero D, Gallo E, Boccadoro M
Dipartimento di Medicina e Oncologia Sperimentale, Universita' di Torino, Ospedale Maggiore S. Giovanni Battista, Italy.
Br J Haematol. 1994 Jul;87(3):494-502. doi: 10.1111/j.1365-2141.1994.tb08303.x.
T cells in multiple myeloma (MM) patients are highly susceptible to activation with the anti-CD3 monoclonal antibody (mAb) OKT3. When short-term OKT3 stimulation is carried out on bone marrow mononuclear cells (BMMC), large numbers of CD3+ CD25+ HLA-DR+ cells are rapidly generated and autologous malignant plasma cells are killed. OKT3 may thus be exploited in autologous bone marrow transplantation (ABMT) to purge residual plasma cells and simultaneously activate T cells to induce graft-versus-leukemia-like (GVL-like) activity upon reinfusion. However, the possible impact of ex-vivo short-term OKT3 stimulation on haematological recovery is unknown. The aim of this work was to investigate the effect of OKT3 stimulation in vitro on autologous haemopoietic progenitor cells (HPC) of MM patients. Colony formation by granulocyte-macrophage progenitor cells (granulocyte-macrophage colony-forming units, CFU-GM) was highly suppressed, although supernatants of OKT3-activated T cells contained up to 2,500 pg/ml of granulocyte-macrophage colony-stimulating factor (GM-CSF). T cell depletion completely prevented this suppression. Neutralizing antibodies against TNF-alpha, TNF-beta and IFN-gamma (which are also produced by OKT3-activated MM T cells) did not prevent it, and Transwell cultures showed that cell-to-cell contact was the main mechanism involved. OKT3-activated T cells also suppressed erythroid burst-forming units (BFU-E) and CFU-GM generation from HPC responsible for long-term maintenance of in vitro myelopoiesis. When tested on normal allogeneic BM, MM supernatants of OKT3-stimulated BMMC partially suppressed the generation of day 7 CFU-GM, but had no effect on day 14 CFU-GM. These data indicate that short-term stimulation of BMMC with OKT3 can be used to generate anti-tumour effector T cells for autologous adoptive immunotherapy. It is not a feasable approach for ex-vivo purging and activation procedures in ABMT because of its potent inhibition of autologous haemopoiesis.
多发性骨髓瘤（MM）患者的T细胞极易被抗CD3单克隆抗体（mAb）OKT3激活。对骨髓单个核细胞（BMMC）进行短期OKT3刺激时，会迅速产生大量CD3 + CD25 + HLA-DR + 细胞，并且自体恶性浆细胞会被杀死。因此，OKT3可用于自体骨髓移植（ABMT），以清除残留的浆细胞，并在重新输注时同时激活T细胞以诱导移植物抗白血病样（GVL样）活性。然而，体外短期OKT3刺激对血液学恢复的可能影响尚不清楚。这项工作的目的是研究体外OKT3刺激对MM患者自体造血祖细胞（HPC）的影响。尽管OKT3激活的T细胞的上清液中含有高达2500 pg/ml的粒细胞-巨噬细胞集落刺激因子（GM-CSF），但粒细胞-巨噬细胞祖细胞（粒细胞-巨噬细胞集落形成单位，CFU-GM）的集落形成受到高度抑制。T细胞清除完全阻止了这种抑制作用。针对肿瘤坏死因子-α、肿瘤坏死因子-β和干扰素-γ（它们也由OKT3激活的MM T细胞产生）的中和抗体并不能阻止这种抑制作用，并且Transwell培养表明细胞间接触是主要的作用机制。OKT3激活的T细胞还抑制了负责体外骨髓生成长期维持的HPC的红系爆式集落形成单位（BFU-E）和CFU-GM的生成。当在正常异基因骨髓上进行测试时，OKT3刺激的BMMC的MM上清液部分抑制了第7天CFU-GM的生成，但对第14天CFU-GM没有影响。这些数据表明，用OKT3对BMMC进行短期刺激可用于产生抗肿瘤效应T细胞用于自体过继免疫治疗。由于其对自体造血的强烈抑制作用，它不是ABMT中体外清除和激活程序的可行方法。
