Dendritic cells transfected

Kyte JA, Mu L, Aamadal S, Kvalheim G, et al. 2006. Phase I/II trial of melanoma therapy with dendritic cells transfected with tumor-RNA. Cancer Gene Ther. 13 905-918. 

Amano T, Kajiwara K, Yoshikawa K, Morioka J, Nomura S, Fujisawa H et al. Antitumor effects of vaccination with dendritic cells transfected with modified receptor for hyaluronan-mediated motility mRNA in a mouse glioma model. J Neurosurg 2007 106 638-645. 

S. S. Diebold, H. Lehrmann, M. Kursa, E. Wagner, M. Cotten, and M. Zenke, Efficient gene delivery into human dendritic cells by adenovirus polyethyleni-mine and mannose polyethylenimine transfection, Hum. Gene Ther., 10 (1999) 775-786. 

COS-7 or CHO cells (for initial transfection screening) or cells of therapeutic interest (e.g., dendritic cells and various cancer cells) at a confluence of 50%, grown in 96-well culture plates, were placed into the robot by the robotic conveyor. In a fully automated process, the robot removes the lid from the cell culture microtiter plate, dispenses lipoplexes into the wells (triplicates), replaces the lid and returns the plate to the incubator. After four hours, the cells are automatically retrieved, the cell monolayers are carefully washed using a special drop mode of the integrated plate washer, fresh medium is added, and the cells are incubated for further 42 hours before harvesting. 

For testing the transfection properties of KL-1-14 toward the mamma carcinoma cell lines MDA-MB-468 and MCF-7, the polarized cell line MDCK-C7, and the primary dendritic cells KL-1-14 was used in its racemic form with chloride as counterion and as a mixture with 60 mol% Choi. In comparison to the transfection efficiencies found for COS-7-cells (see above), the transfection efficiencies were generally lower (Table 2). For MDA-MB-468 and MCF-7, transfection efficiencies were reduced by a factor of about 10, the MDCK-C7-cells by a factor of about 80, and the dendritic cells by a factor of about 500. Nevertheless, the transfection efficiencies found for KL-l-14/Chol (1 0.6) were generally higher than for DOTAP, respectively. For the mamma carcinoma cell lines, transfection efficiencies with KL-1-14/ Choi (1 0.6) were four times higher than for DOTAP. For MDCK-C7 and dendritic cells, the increase was from 1.9- to 2.5-fold. We also tested KL-1-14 as equimolar mixture with DOPE for its transfection efficiencies toward the mamma carcinoma cell lines and the dendritic cells. Again, transfection efficiencies were greatly reduced even for the KL-1-14/DOPE mixture and were similar to the values found for KL-l-14/Chol (1 0.6). 

Table 2 Transfection Efficiencies of KL-1-14 in a Mixture with Choi (Ratio 0.7) or DOPE (Ratio 1.0) Toward the Mamma Carcinoma Cell Lines MDA-MB-468 and MCE-7, the Polarized Cell Line MDCK-C7, and the Primary Dendritic Cells...

Abbreviations DOPE, l,2-dioleoyl-sn-glycero-3-phosphatidylethanolamine DOTAP, iV-[l-(2, 3-dioleoyloxy)propyl]-A,iV,JV-trrmethylanimoniumchloride TE, transfection efficiency DC, dendritic cells RLU, relative light units chol, cholesterol. 

Irvine AS, Trinder PKE, Laughton DL, et al. Efficient nonviral transfection of dendritic cells and their use for in vivo immunisation. Nat Biotechnol 2000 18 1273-8. 

Drury AN, Szent-Gyorgyi A (1929) The physiological activity of adenine compounds with especial reference to their action upon the mammalian heart. J Physiol 68(3) 213—237 Englert M, Quitterer U, Klotz K-N (2002) Effector coupling of stably transfected human A3 adenosine receptors in CHO cells. Biochem Pharmacol 64(l) 69-73 Fossetta J, Jackson J, Deno G, Fan X, Du XK, Bober L, Soude-Bermejo A, de Bouteiller O, Caux C, Lunn C, Lundell D, Palmer RK (2003) Pharmacological analysis of calcium responses mediated by the human A, adenosine receptor in monocyte-derived dendritic cells and recombinant cells. Mol Pharmacol 63(2) 342-350... 

Porgador, A., Irvine, K.R., Iwasaki, A., Barber, B.H., Restifo, N.P. and Germain, R.N. (1998) Predominant role for directly transfected dendritic cells in antigen presentation to CD8+ T cells after gene gun immunization. J. Exp. Med., 188, 1075-1082. 

Skin is rich in dendritic cells, which are potent initiators of immune responses and possess the co-stimulatoiy and adhesion molecules required for T cell activation. In addition, dendritic cells possess a unique ability to process and present extracellular antigens in the context of both class I and class II molecules. Thus, transfection of plasmids into these cells is likely to elicit both cellular and humoral responses. Specific targeting of dendritic cells residing in the lymph nodes will likely represent an attractive strategy for providing a robust immune response with nucleic acid vaccines. 

In wVo transfection of dendritic cells with liposomal plasmid-DNA... 

Su Z, Dannull J, Yang BK, Dahm P, Coleman D, Yancey D, Sichi S, Niedzwiecki D, Boczkowski D, Gilboa E, Vieweg J. Telomerase mRNA-transfected dendritic cells stimulate antigen-specific CD8 and CY>4 T cell responses in patients with metastatic prostate cancer. J Immunol 2005 174 3798-3807. 

Cancer immunotherapy by genetically engineered dendritic cells that have been transfected witli genes encoding tumor-associated antigens provides an intriguing combination of gene tlierapy and immunotherapy . ... 

Alistair S. Irvine, Peter K. E. Trinder, David L. Laughton, Helen Ketteringham, Ruth H. McDermott, Sophie C. H. Reid, Adrian M. R. Haines, Abdu Amir, Rhonda Husain, Rajeev Doshi, Lawrence S. Young, Andrew Mountain, Efficient nonviral transfection of dendritic cells and their use for in vivo immunization. Nature Biotechnology, 18 (2000), 1273-1278. 

Mockey M, Goncalves C, Dupuy P P, et al. (2006). mRNA transfection of dendritic cells synergistic effect of ARCA mRNA capping with Poly(A) chains in cis and in trans for a high protein expression level. Biochem. Biophys. Res. Commun. 340 1062-1068. 

Gilboa E, Vieweg J (2004). Cancer immunotherapy with mRNA-transfected dendritic cells. Immunol. Rev. 199 251-263. 

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