《Kras突变多肽与全细胞抗原致敏DCs诱导CTLs对胰腺癌的杀伤活性研究》.pdfVIP

Chinese-German Journal of Clinical Oncology December 2010, Vol. 9, No. 12, P724–P729 DOI 10.1007/s10330-010-0707-1 Anti-tumor effect of CTLs activated by dendritic cells pulsed with K-ras mutant peptide and whole tumor antigen on pancreatic cancer* Guang Tan, Zhongyu Wang, Xin Zhang, Zhengang Cai, Junkai Zhang Department of General Surgery, The First Affiliated Hospital of Dalian Medical University, Dalian 116011, China Received: 14 September 2010 / Revised: 20 October 2010 / Accepted: 5 November 2010 © Huazhong University of Science and Technology and Springer-Verlag Berlin Heidelberg 2010 Abstract Objective: We studied the role of specific cytotoxic T lymphocytes (CTLs) activated by dendritic cells (DCs) presenting cationic nanoparticles with the K-ras (12-Val) mutant peptide and whole tumor antigen in the killing of different pancreatic cancer cell lines in vitro and in vitro. Methods: Peripheral blood DCs were induced by rhGM-CSF and IL-4 and cul- tured. DCs were sensitized by whole antigen of a pancreatic cancer cell line (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 ELISA was performed to detect IFN-γ secretion. 125I-UdR was used to measure the killing effect of CTLs. We also evaluated the antitumor activity of CTLs in vivo in a tumor-bearing nude mouse model prepared with the PANC-1 (K-ras+) and SW1990 (K-ras–) cell lines. Results: Compared with K-ras+peptide, low concentration K-ras+peptide-CNP can be effectively presented by DCs (P 0.05). CTLs induced by