Immunolipoplexes: an efficient, nonviral alternative for transfection of human dendritic cells with potential for clinical vaccination.
Genetic manipulation of dendritic cells (DCs) is important in the context of using either mature DCs to immunize patients or immature DCs to induce tolerance. Here, we describe a novel method of transfecting monocyte-derived human DCs using immunolipoplexes containing anti-CD71 or anti-CD205 monoclonal Abs. This results in up to 20% transfection, which can be increased to 20-30% if the immunolipoplexes are used to transfect CD14+ monocytes prior to differentiation into DCs. Transfected DCs can be substantially enriched using a drug-selection protocol during differentiation. Unlike adenoviral transduction, this nonviral transfection does not alter the expression of costimulatory molecules or the production of proinflammatory cytokines by DCs. In addition, DC function is unaltered, as assessed by mixed lymphocyte reactions. To test the feasibility of the immunolipoplexes and selection protocol for therapeutic intervention, we transfected DCs with the immunomodulatory enzyme indoleamine 2,3-dioxygenase (IDO). Allogeneic T cells exposed to IDO-expressing DCs did not proliferate, secreted more IL-10 and less Th1 and Th2 cytokines, and had a higher amount of apoptosis than T cells incubated with control DCs. Furthermore the remaining T cells were rendered anergic to further stimulation by allogeneic DC. These immunolipoplexes, which can be easily and rapidly assembled, have potential for clinical immunization, in particular for tolerance induction protocols.
Tan
Peng H
PH
Department of Immunology, Division of Medicine, Faculty of Medicine, Imperial College London, Hammersmith Hospital, Du Cane Road, London W12 ONN, UK.
Beutelspacher
Sven C
SC
Wang
Yao-He
YH
McClure
Myra O
MO
Ritter
Mary A
MA
Lombardi
Giovanna
G
George
Andrew J T
AJ
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D015703:Antigens, CD / Q000276:immunology; D000944:Antigens, Differentiation, B-Lymphocyte / Q000276:immunology; D002454:Cell Differentiation; D002478:Cells, Cultured; D018920:Coculture Techniques; D003713:Dendritic Cells / Q000166:cytology / Q000276:immunology* / Q000378:metabolism* / Q000637:transplantation; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D050503:Indoleamine-Pyrrole 2,3,-Dioxygenase; D037181:Lectins, C-Type / Q000276:immunology; D008081:Liposomes / Q000008:administration & dosage* / Q000276:immunology; D051379:Mice; D015778:Minor Histocompatibility Antigens; D010641:Phenotype; D011956:Receptors, Cell Surface / Q000276:immunology; D011990:Receptors, Transferrin; D013601:T-Lymphocytes / Q000276:immunology / Q000378:metabolism; D014162:Transfection / Q000295:instrumentation*; D014366:Tryptophan Oxygenase / Q000235:genetics / Q000378:metabolism; D014611:Vaccination / Q000379:methods*
D015703:Antigens, CD; D000944:Antigens, Differentiation, B-Lymphocyte; C068126:CD71 antigen; C093569:DEC-205 receptor; D050503:Indoleamine-Pyrrole 2,3,-Dioxygenase; D037181:Lectins, C-Type; D008081:Liposomes; D015778:Minor Histocompatibility Antigens; D011956:Receptors, Cell Surface; D011990:Receptors, Transferrin; D014366:Tryptophan Oxygenase