Accelerated co-cultured dendritic cell (acDC) loaded with autologous apoptotic bodies might be a promising approach for antigen delivery

K. Kalantar, N. Gholijani, E. Martinuzzi, S. Culina, D. Kabelitz, Z. Amirghofran
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Abstract

ABSTRACT Antigens derived from engulfed apoptotic bodies that are presented by dendritic cells can amplify Ag-specific T-cells. Accelerated co-cultured DC (acDC) strategy keeps lymphocytes in contact with differentiating DCs. Therefore, Ag-specific T-cell activation can occur during DC maturation. Our aim was to prepare DCs by acDC method and check the subsequent engulfment of the apoptotic body by acDC. We have proposed that this method could be feasible if we transfect the apoptotic bodies with the antigen. DCs were prepared using acDC method and their maturation markers were confirmed by flow cytometry. Ultraviolet was used for inducing apoptosis in the PBMCs and induction of apoptosis checked by propidium iodide and 7-aminoactinomycin D staining. Flow cytometry and immunohistochemistry were used for checking the uptake of apoptotic bodies by the DCs. The alloreactivity against apoptotic bodies was examined by enzyme-linked immunospot (ELISPOT) assay. Results showed that 40.4% of DCs could efficiently engulf the apoptotic bodies. The results indicated that acDC method is capable to isolate a high yield of DCs, and these cells could properly engulf the apoptotic bodies, more works should be performed to use this method for Ag discovery through delivering the Ag by apoptotic bodies into the DCs.
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负载自体凋亡小体的加速共培养树突状细胞(acDC)可能是一种很有前途的抗原递送方法
树突状细胞呈递的来自吞噬凋亡小体的抗原可以扩增ag特异性t细胞。加速共培养DC (acDC)策略保持淋巴细胞与分化的DC接触。因此,ag特异性t细胞激活可以在DC成熟过程中发生。我们的目的是用acDC法制备dc,并观察acDC对凋亡小体的吞噬情况。我们已经提出,如果用抗原转染凋亡小体,这种方法是可行的。采用acDC法制备树突状细胞,流式细胞术鉴定其成熟标志物。紫外诱导PBMCs凋亡,碘化丙啶染色和7-氨基放线菌素D染色检测凋亡诱导情况。流式细胞术和免疫组织化学检测树突状细胞对凋亡小体的摄取。采用酶联免疫斑点法(ELISPOT)检测其对凋亡小体的异体反应性。结果表明,40.4%的树突状细胞能有效吞噬凋亡小体。结果表明,acDC方法能够高产量地分离出dc细胞,并且这些细胞能够很好地吞噬凋亡小体,利用这种方法通过凋亡小体将Ag传递到dc细胞中发现Ag还需要做更多的工作。
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