Siena S, Di Nicola M, Bregni M, Mortarini R, Anichini A, Lombardi L, Ravagnani F, Parmiani G, Gianni A M
Cristina Gandini Transplantation Unit, Istituto Nazionale Tumori, Milan, Italy.
Exp Hematol. 1995 Dec;23(14):1463-71.
We report that blood cell autografts, collected by single leukapheresis in cancer patients (n = 11) at the time of mobilization of hematopoietic progenitors into peripheral blood following anticancer therapy with high-dose cyclophosphamide (HD-CTX) plus interleukin-3 (IL-3) and granulocyte colony-stimulating factor (G-CSF/filgrastim), comprise 1.98 +/- 0.39 x 10(5)/kg (mean +/- SE) CD34+ progenitors of dendritic cells (DCs). This number corresponds to 140-fold more progenitors than in a control autograft collected in the steady state. DCs derived from mobilized CD34+ cells, morphologically and immunophenotypically undistinguishable from skin Langerhans cells and DCs from bone marrow and cord blood CD34+ cells, are shown to be powerful stimulators of allogeneic T cell proliferation in primary MLR and of autologous HLA-DR-restricted CD4+ T cell proliferation in response to presentation of xenogenic antigens. We show that the GM-CSF-plus-TNF-alpha-dependent ex vivo generation of DCs from mobilized CD34+ cells is 2.5-fold enhanced by flk-2/flt-3 ligand or c-kit ligand (stem cell factor) and five-fold enhanced by a combination of these growth factors. In addition, the optimal serum for the generation of DCs is autologous HD-CTX recovery-phase serum rather than fetal calf serum (FCS) or steady-state human serum, which are clinically inadequate and ineffective, respectively. In practice, the stimulation of CD34+ cells in a blood cell autograft (15.75 +/- 2.46 x 10(6)/kg) provided by the above four growth factors should permit ex vivo generation of approximately 40 x 10(9) DCs in an adult patient. These new findings provide advantageous tools for the large-scale generation of DCs that are potentially usable for clinical protocols of immunotherapy or vaccination in patients undergoing cancer treatment.
我们报告，在癌症患者（n = 11）接受大剂量环磷酰胺（HD-CTX）加白细胞介素-3（IL-3）和粒细胞集落刺激因子（G-CSF/非格司亭）进行抗癌治疗后，造血祖细胞动员至外周血时，通过单次白细胞分离术采集的血细胞自体移植物中，包含1.98±0.39×10⁵/kg（均值±标准误）的树突状细胞（DCs）CD34⁺祖细胞。这个数量比在稳态下采集的对照自体移植物中的祖细胞多140倍。源自动员的CD34⁺细胞的DCs，在形态学和免疫表型上与皮肤朗格汉斯细胞以及来自骨髓和脐血CD34⁺细胞的DCs无法区分，在原发性混合淋巴细胞反应（MLR）中显示为同种异体T细胞增殖的强大刺激剂，在响应异种抗原呈递时为自体HLA-DR限制性CD4⁺T细胞增殖的刺激剂。我们表明，flk-2/flt-3配体或c-kit配体（干细胞因子）可使从动员的CD34⁺细胞体外生成GM-CSF加TNF-α依赖的DCs增加2.5倍，而这些生长因子的组合可使其增加5倍。此外，生成DCs的最佳血清是自体HD-CTX恢复期血清，而非胎牛血清（FCS）或稳态人血清，后两者在临床上分别不足且无效。实际上，上述四种生长因子对血细胞自体移植物中CD34⁺细胞（15.75±2.46×10⁶/kg）的刺激应能使成年患者体外生成约40×10⁹个DCs。这些新发现为大规模生成DCs提供了有利工具，这些DCs可能可用于癌症治疗患者的免疫治疗或疫苗接种临床方案。
