Previously, we have shown platelet lysate (PL) can be used as a non-xenogeneic serum supplement for generation of monocyte-derived dendritic cells (DCs). Since DC-based activation protocols are extremely sensitive to microenvironmental changes such as replacement of culture medium, we wanted to examine the behavior of DCs cultured in the presence of PL under various type-1 activation conditions and assess their type 1 polarization capacity. We compared the quality of DCs cultured in 10% PL-supplemented RPMI medium (plDCs) with clinical-grade DCs obtained using commercially available serum-free medium (sfDCs), frequently used in established DC vaccine protocols. The DC maturation protocols consisted of either monophosphoryl lipid A/IFN-γ, poly I:C/TNF-α/IFN-α or poly I:C/R848. In general, plDCs were inferior to sfDCs in most aspects of their functional type 1 polarization characteristics. After maturation, the expression of co-stimulatory, HLA class II and lymph node-homing molecules was strongly up-regulated, with some noticeable differences. The expression of CD80 and CD86 was more extensive on plDCs, which was particularly evident in case of CCR7. However, after observing their functional capacity, plDCs had significantly lower allo-stimulatory capacity both in terms of CD4+ and CD8+ T cell stimulation. The high expression of CCR7 corresponded to higher CCL-19 directed DC migration of plDCs compared to sfDCs. Finally, their capacity to induce granzyme B and IFN-γ production in CD8+ T cells was significantly reduced in comparison to sfDCs. Based on these findings, the use of PL as an alternative serum supplement for generation of monocyte-derived DC anti-tumor vaccines is questionable. Abbreviations: Ag: antigen; CCL: chemokine ligand; CCR: chemokine receptor; DC: dendritic cells; DC-SIGN: dendritic cell-specific intercellular adhesion molecule-3-grabbing non-integrin; FBS: fetal bovine serum; GMP: good manufacturing practice; IFN: interferon; IL: interleukin; MPLA: monophosphoryl lipid A; PGE: prostaglandin E; pI:C: polyinosinic:polycytidylic acid; pl: platelet lysate; sf: serum free; TLR: toll-like receptor; TNF: tumor necrosis factor.

中文翻译：

---

存在血小板溶解产物的树突状细胞具有降低的1型极化能力。

以前，我们已经显示血小板裂解物（PL）可用作非异种血清补充剂，用于产生单核细胞衍生的树突状细胞（DC）。由于基于DC的激活协议对微环境的变化（例如培养基的更换）极为敏感，因此，我们希望检查在PL存在下在各种1型激活条件下培养的DC的行为，并评估其1型极化能力。我们将10％PL补充的RPMI培养基（plDC）中培养的DC与使用市售无血清培养基（sfDCs）获得的临床级DC的质量进行了比较，后者通常用于已建立的DC疫苗方案中。DC成熟方案由单磷酰基脂质A /IFN-γ，聚I：C /TNF-α/IFN-α或聚I：C / R848组成。一般来说，plDC在其功能性1型极化特性的大多数方面均次于sfDC。成熟后，共刺激，HLA II类和淋巴结归巢分子的表达被强烈上调，但有一些明显的差异。CD80和CD86在plDC上的表达更为广泛，在CCR7的情况下尤其明显。但是，观察到它们的功能能力后，就DC4 +和CD8 + T细胞刺激而言，plDC的同种异体刺激能力明显较低。与sfDC相比，CCR7的高表达对应于plDC的更高的CCL-19定向DC迁移。最后，与sfDC相比，它们诱导CD8 + T细胞中粒酶B和IFN-γ产生的能力大大降低。根据这些发现，使用PL作为产生单核细胞的DC抗肿瘤疫苗的替代血清补充剂存在疑问。缩写：Ag：抗原；CCL：趋化因子配体；CCR：趋化因子受体；DC：树突状细胞；DC-SIGN：树突状细胞特异的细胞间粘附分子-3-非整合素。FBS：胎牛血清；GMP：良好的生产规范；干扰素：干扰素；IL：白介素；MPLA：单磷酰基脂质A；PGE：前列腺素E；pI：C：聚肌苷酸：聚胞苷酸；pl：血小板裂解物；sf：无血清；TLR：收费型受体；TNF：肿瘤坏死因子。良好生产规范; 干扰素：干扰素；IL：白介素；MPLA：单磷酰基脂质A；PGE：前列腺素E；pI：C：聚肌苷酸：聚胞苷酸；pl：血小板裂解物；sf：无血清；TLR：收费型受体；TNF：肿瘤坏死因子。良好生产规范; 干扰素：干扰素；IL：白介素；MPLA：单磷酰基脂质A；PGE：前列腺素E；pI：C：聚肌苷酸：聚胞苷酸；pl：血小板裂解物；sf：无血清；TLR：收费型受体；TNF：肿瘤坏死因子。