Zoology Department, Faculty of Science, Tanta University, El-Giesh St., Gharbia province, Tanta 31527, Egypt; Center of Excellence in Cancer Research, Tanta University, El-Giesh St., Gharbia province, Tanta 31527, Egypt.
Zoology Department, Faculty of Science, Tanta University, El-Giesh St., Gharbia province, Tanta 31527, Egypt.
Biomed Pharmacother. 2017 Sep;93:286-295. doi: 10.1016/j.biopha.2017.06.046. Epub 2017 Jun 23.
In order to get mature dendritic cells (DC) that is a crucial prerequisite for success in tumor immunotherapy protocols. Herein, we assumed that administration of murine bone marrow (BM)-derived DC (BM-DC), loaded ex vivo with whole Ehrlich ascites carcinoma (EAC) lysate, in the context of systemic chemotherapy cyclophosphamide (CTX) to induce antitumor immune responses, may be a good strategy to improve the presentation of tumor-specific antigens to the immune system. In the first series of experiments, BM cells generated either from BM of naïve mice or from BM of EAC-bearing mice were cultured in the presence of GM-CSF and IL-4 for 6days. At day 7, cells were loaded for 48h with one of the following maturation agents: EAC lysate (1mg/ml), poly-inosinic: polycytidylic acid [poly(I:C)] (25μg/ml) or mRNA encoding human telomerase reverse transcriptase (hTERT-mRNA) (2μg/ml). In the second series of experiments, EAC-bearing mice were intraperitoneally (i.p.) injected with CTX followed by i.p. vaccination with DC, loaded ex vivo with EAC lysate. DC yield and the phenotypic expression of maturity-related surface markers of DC (i.e. CD11b and CD11c) in both series of experiments were investigated. As a result, a significant decrease in the number of DC generated from poly(I:C)-supplemented BM culture from EAC-bearing mice has been detected. Loading of BM cells with poly(I:C), EAC lysate or hTERT-mRNA could induce the expression of CD11b and CD11c. Additionally, vaccination of EAC-bearing mice with DC loaded ex vivo with EAC lysate following CTX treatment, resulted in increases in the percentage of multiple populations of CD11bCD11c in BM, spleen and peripheral blood (PB). To conclude, further researches to clarify the mechanism involved in DC maturation are crucial not only to comprehend DC biology but also to optimize DC immunotherapy protocols.
为了获得成熟的树突状细胞（DC），这是肿瘤免疫治疗方案成功的关键前提。在这里，我们假设在全身化疗环磷酰胺（CTX）的背景下，给予骨髓（BM）来源的负载整个艾氏腹水癌（EAC）裂解物的 DC（BM-DC），可以诱导抗肿瘤免疫反应，这可能是一种改善肿瘤特异性抗原向免疫系统呈递的好策略。在第一组实验中，从 naive 小鼠或 EAC 荷瘤小鼠的 BM 中生成的 BM 细胞在 GM-CSF 和 IL-4 的存在下培养 6 天。在第 7 天，用以下成熟剂之一负载细胞 48 小时：EAC 裂解物（1mg/ml）、聚肌苷酸：聚胞苷酸[poly(I:C)]（25μg/ml）或编码人端粒酶逆转录酶（hTERT-mRNA）的 mRNA（2μg/ml）。在第二组实验中，EAC 荷瘤小鼠腹腔内（i.p.）注射 CTX 后，腹腔内接种负载 EAC 裂解物的 DC。在这两组实验中，都研究了 DC 的产量和成熟相关表面标志物的表型表达（即 CD11b 和 CD11c）。结果，从 EAC 荷瘤小鼠的 poly(I:C)补充 BM 培养物中生成的 DC 数量显著减少。用 poly(I:C)、EAC 裂解物或 hTERT-mRNA 负载 BM 细胞可以诱导 CD11b 和 CD11c 的表达。此外，在 CTX 处理后，用负载 EAC 裂解物的 DC 对 EAC 荷瘤小鼠进行疫苗接种，导致 BM、脾脏和外周血（PB）中多个 CD11bCD11c 群体的百分比增加。总之，进一步研究阐明 DC 成熟涉及的机制不仅对于理解 DC 生物学至关重要，而且对于优化 DC 免疫治疗方案也至关重要。
