Hubert P, Greimers R, Franzen-Detrooz E, Doyen J, Delanaye P, Boniver J, Delvenne P
Department of Pathology, CHU Sart Tilman, Liege, Belgium.
Cancer Immunol Immunother. 1998 Oct;47(2):81-9. doi: 10.1007/s002620050507.
Dendritic cells (DC) are the most efficient antigen presenting cells. The clinical use of DC as vectors for antitumor and anti-infectious disease immunotherapy has been limited by their low level and accessibility in normal tissue. Substantial numbers of DC can be generated from peripheral blood cultured in the presence of interleukin-4 (IL-4) and granulocyte/macrophage-colony-stimulating factor (GM-CSF). We showed in this study that substantial numbers of DC can be obtained from the peripheral blood of patients with (pre)neoplastic lesions of the uterine cervix. The procedure required relatively small blood samples (10 ml) and the presence of 100 U/ml IL-4 and 800 U/ml GM-CSF in the culture medium. There was no significant difference in the morphology, yield, phenotype and function of generated DC between patients with cervical (pre)neoplastic lesions and healthy individuals. When the hematopoietic factor Flt3 ligand (Flt3L, 40 ng;ml) was added, there was an average increase in the DC population of 26% compared to cultures with GM-CSF and IL-4 alone. Approximately 1.2 x 10(6) cells with the characteristics of dendritic cells could be obtained when Flt3L was included in the medium. The addition of Flt3L did not modify the phenotypic profile of DC (HLA-DR+, CD1a+, CD4+, CD54+, CD80+, CD86+. CD40+, CD3- and CD14-). In addition, Flt3L generated functional DC capable of stimulating the proliferation of alloreactive T cells. These results suggest that Flt3L, in association with GM-CSF and IL-4, provides an advantageous tool for the large-scale generation of DC and that an immunotherapy based on the use of DC generated in vitro is possible in patients with (pre)neoplastic lesions of the uterine cervix.
树突状细胞（DC）是最有效的抗原呈递细胞。DC作为抗肿瘤和抗感染疾病免疫治疗载体的临床应用，一直受到其在正常组织中含量低且难以获取的限制。在白细胞介素-4（IL-4）和粒细胞/巨噬细胞集落刺激因子（GM-CSF）存在的情况下培养外周血，可产生大量DC。我们在本研究中表明，可从子宫颈（癌前）病变患者的外周血中获得大量DC。该过程所需的血样相对较少（10毫升），且培养基中需含有100 U/ml的IL-4和800 U/ml的GM-CSF。子宫颈（癌前）病变患者与健康个体所产生的DC在形态、产量、表型和功能方面无显著差异。当添加造血因子Flt3配体（Flt3L，40 ng/ml）时，与单独使用GM-CSF和IL-4的培养物相比，DC群体平均增加了26%。当培养基中含有Flt3L时，可获得约1.2×10⁶个具有树突状细胞特征的细胞。添加Flt3L并未改变DC的表型特征（HLA-DR⁺、CD1a⁺、CD4⁺、CD54⁺、CD80⁺、CD86⁺、CD40⁺、CD3⁻和CD14⁻）。此外，Flt3L产生的功能性DC能够刺激同种异体反应性T细胞的增殖。这些结果表明，Flt3L与GM-CSF和IL-4联合使用，为大规模产生DC提供了一种有利工具，并且基于体外产生的DC的免疫治疗在子宫颈（癌前）病变患者中是可行的。
