buy FK228 PubMedCrossRef 32. Shafer-Weaver K, Rosenberg S, Strobl S, Gregory Alvord W, Baseler M, Malyguine selleck A: Application of the granzyme B ELISPOT assay for monitoring cancer vaccine trials. J Immunother 2006, 29:328–335.PubMedCrossRef
33. Makedonas G, Banerjee PP, Pandey R, Hersperger AR, Sanborn KB, Hardy GA, Orange JS, Betts MR: Rapid up-regulation and granule-independent transport of perforin to the immunological synapse define a novel mechanism of antigen-specific CD8+ T cell cytotoxic activity. J Immunol 2009, 182:5560–5569.PubMedCrossRef 34. Snyder-Cappione JE, Divekar AA, Maupin GM, Jin X, Demeter LM, Mosmann TR: HIV-specific cytotoxic cell frequencies measured directly ex vivo by the Lysispot assay can be higher or lower than the frequencies of IFN-gamma-secreting cells: anti-HIV cytotoxicity is not generally impaired relative to other chronic virus responses. see more J Immunol 2006, 176:2662–2668.PubMed 35. Khan IF, Hirata RK, Russel DW: AAV-mediated gene targeting
methods for human cells. Nat Protocols 2011, 6:482–501.CrossRef 36. Yin Y, Cao LY, Wu WQ, Li H, Jiang Y, Zhang HF: Blocking effects of siRNA on VEGF expression in human colorectal cancer cells. World J Gastroenterol 2010,16(9):1086–1092.PubMedCrossRef 37. Zhang X, Ge YL, Tian RH: The knockdown of c-myc expression by RNAi inhibits cell proliferation in human colon cancer HT-29 cells in vitro and in vivo. Cell Mol Biol Lett 2009,14(2):305–318.PubMedCrossRef 38. Li H, Cao HF, Wan J, Li Y, Zhu ML, Zhao P: Growth inhibitory effect of wild-type Kras2 gene on a colonic adenocarcinoma cell line. World J Gastroenterol 2007,13(6):934–938.PubMedCrossRef 39. Cao J, Yu JP, Liu CH, Zhou L, Yu HG: Effects of gastrin 17 on beta-catenin/Tcf-4 pathway in Colo320WT colon cancer cells. World J Gastroenterol 2006,12(46):7482–7487.PubMed
40. Dalby B, Cates S, Harris A, Ohki EC, Tilkins ML, Price PJ, Ciccarone VC: Advanced transfection with Lipofectamine 2000 reagent: primary neurons, siRNA, and high-throughput applications. selleck chemicals Methods 2004,33(2):95–103.PubMedCrossRef 41. Yamano S, Dai J, Morsi AM: Comparison of trasfection efficiency of non viral gene transfer reagent. Molecular Biotechnol 2010, 46:287–300.CrossRef 42. Monsurrò V, Nagorsen D, Wang E, Provenzano M, Dudley ME, Rosenberg SA, Marincola FM: Functional heterogeneity of vaccine-induced CD8(+) T cells. J Immunol 2002, 168:5933–5942.PubMed Competing interests There are no competing interests (political, personal, religious, ideological, academic, intellectual, commercial or any other) to declare in relation to this manuscript by all authors. Authors’ contributions VB, AC, PCF carried out the immunoassays and participated in the design of the study and performed the statistical analysis. MR and ES carried out the transfection protocol. MZ supplied the cells from the animal model. VB, GR PCF FE helped to draft the manuscript. MPF conceived of the study, and participated in its design and coordination and helped to draft the manuscript.