Induction of CTLs by DCs pulsed with K-ras mutant peptide on the surface of nanoparticles in the treatment of pancreatic cancer.
Induction of CTLs by DCs pulsed with K-ras mutant peptide on the surface of nanoparticles in the treatment of pancreatic cancer.
复制标题
在胰腺癌的治疗中，通过纳米粒子表面的 K-ras 突变肽脉冲，DC 诱导 CTL。
DOI：
10.3892/or.2011.1283
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发表时间：
2011-07
期刊：
影响因子：
4.2
通讯作者：
中科院分区：
文献类型：
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作者：
本研究的目的是研究树突状细胞（DCs）提呈K-ras（12-瓦尔）突变肽阳离子纳米颗粒后，其激活的特异性细胞毒性T淋巴细胞（CTL）对胰腺癌细胞株的杀伤作用。用rhGM-CSF和IL-4诱导培养外周血DC。分别用PANC-1全抗原、K-ras突变肽（K-ras +peptide）和K-ras突变肽阳离子纳米粒（K-ras+ peptide-CNP）致敏DC。通过流式细胞术测量细胞表面标志物。3 H-TdR法检测淋巴细胞增殖反应，ELISA法检测IL-12和IFN-γ的分泌。用125 I-UdR检测CTL的杀伤作用。在PANC-1和SW 1990细胞制备的荷瘤裸鼠模型中评价CTL的抗肿瘤活性。结果显示，与K-ras+肽相比，低浓度的K-ras+肽-CNP可被DC有效提呈（P<0.05）。与K-ras+肽CNP和K-ras+肽CNP诱导的CTL相比，全瘤抗原致敏的DC诱导的CTL对PANC-1和SW 1990胰腺癌细胞的杀伤作用显著增强（P<0.05）。K-ras+肽和K-ras+肽- CNP负载的DC诱导的CTL对PANC-1细胞有特异性杀伤作用（P<0.05），对SW 1990细胞无杀伤作用（P>0.05）。总之，具有K-ras（12-瓦尔）突变肽的阳离子纳米颗粒可以在低浓度下被DC有效地呈递。K-ras+肽-CNP诱导的CTL对K-ras突变的胰腺癌细胞株具有特异性杀伤活性，显著抑制肿瘤生长，延长荷瘤裸鼠的生存时间。虽然本研究证实了全细胞抗原诱导了良好的抗肿瘤免疫应答，但先前已证实的免疫耐受和自身免疫的可能性导致了该DC疫苗应用的困难。
The aim of this study was to investigate the role of specific cytotoxic T lymphocytes (CTLs) activated by dendritic cells (DCs) presenting cationic nanoparticles with the K-ras (12-Val) mutant peptide in the killing of different pancreatic cancer cell lines in vivo and in vitro. Peripheral blood DCs were induced by rhGM-CSF and IL-4 and cultured. DCs were sensitized by whole antigen of PANC-1 with expression of K-ras mutant, K-ras mutant peptide (K-ras+peptide) and cationic nanoparticles with K-ras mutant peptide (K-ras+peptide-CNP), respectively. Cell surface markers were measured by flow cytometry. Lymphocyte proliferation was detected by the 3H-TdR test, and IL-12 and IFN‑γ secretion was detected by ELISA. 125I-UdR was used to measure the killing effect of CTLs. The antitumor activity of CTLs in tumor-bearing nude mouse models prepared with PANC-1 and SW1990 cells was evaluated. Results showed that, compared with K-ras+peptide, low concentrations of K-ras+peptide-CNP were effectively presented by DCs (P<0.05). CTLs induced by DCs pulsed with whole tumor antigen had a significantly greater killing effect (P<0.05) on PANC-1 and SW1990 pancreatic cancer cells compared with K-ras+peptide- and K-ras+peptide-CNP-induced CTLs. CTLs induced by DCs pulsed with K-ras+peptide and K-ras+peptide- CNP had a specific killing effect (P<0.05) on PANC-1 cells and no effect (P>0.05) on SW1990 cells. In conclusion, cationic nanoparticles with the K-ras (12-Val) mutant peptide can be effectively presented by DCs at a low concentration. CTLs induced by K-ras+peptide-CNP had specific killing activity for the pancreatic cancer cell line with the K-ras mutant and significantly inhibited tumor growth and increased the survival time of tumor-bearing nude mice. Although this study confirmed that whole cell antigen induced a good antitumor immune response, the possibility of immune tolerance and autoimmunity which has been previously proven contribute to the difficulty in the application of this DC vaccine.