Immunization of Patients with Malignant Melanoma with Autologous CD34⁺Cell-Derived Dendritic Cells TransducedEx Vivowith a Recombinant Replication-Deficient Vaccinia Vector Encoding the Human Tyrosinase Gene: A Phase I Trial

Boosting T Cell-Mediated Immunity to Tyrosinase by Vaccinia Virus-Transduced, CD34+-Derived Dendritic Cell Vaccination

“…Written informed consent was obtained from each patient before enrolling in the study. Trial design and objectives, as well as eligibility and exclusion criteria, were described extensively elsewhere (18). Briefly, the trial was designed to enroll 6 patients with the primary objective of defining the safety and toxicity of injections of autologous CD34 ϩ -derived dendritic cells transduced ex vivo with a recombinant nonreplicating vaccinia vector encoding the human tyrosinase gene (DCs/MVA-hTyr).…”

“…The yield of CD34 ϩ selected cells, after Clini-MACS processing, was 164 ϫ 10 6 Ϯ 30 ϫ 10 6 with an average purity of 95% Ϯ 2. Purified CD34 ϩ cells were cultured with 10% autologous serum in Iscove's modified Dulbecco's medium (Cambrex, Milan, Italy) supplemented with granulocyte macrophage colony-stimulating factor (50 ng/ml; Myelogen/ Leucomax, Schering-Plough/Sandoz, Milan, Italy), tumor necrosis factor ␣ (10 ng/ml; Knoll, Ludwigshafen, Germany), recombinant human stem cell factor (50 ng/ml; Amgen, Thousand Oaks, CA), and Flk-2/Flt-3 ligand (50 ng/ml; Immunex, Seattle, WA), as described (18 6 cells, resuspended in 50 ml of cold cryoprotectant mixture (90% autologous serum; 10% DMSO) and then frozen into 100-ml polyolefin freezing bags (Delmed, New Brunswick, NJ).…”

Induction of effective therapeutic antitumor immunity by direct in vivo administration of lentiviral vectors

“…[5][6][7] Various methods of loading DC with antigenic peptides, proteins, DNA or mRNA and viral vectors have been established. [8][9][10][11][12][13] Efficient gene delivery to human DC has been achieved with a variety of viral vectors derived from adenovirus, 8,14 adeno-associated virus, 13 onco-retrovirus, 10,11,15 pox viruses such as vaccinia, 9,16 and canary pox 17,18 and lentivirus. [19][20][21][22] Although adenovirus-and vaccinia virus-derived vectors result in efficient transduction and immune activation, they express viral proteins that evoke a potent antivector immune response, which might dampen the immune response against the tumor antigen in repeated vaccination regimens.…”

Effects of Herpes Simplex Virus Amplicon Transduction on Murine Dendritic Cells

“…With their strong capability in antigen presentation, DCs have been employed as natural potent adjuvants for immunization (Mayordomo et al, 1997), pulsed with target peptide(s) (Inaba et al, 1990;Celluzzi et al, 1996), and transduced with various viral vector platforms to efficiently present antigens of interest (Takayama et al, 1999;Di Nicola et al, 2003;Koya et al, 2003Koya et al, , 2004Timares et al, 2004;He et al, 2005). Many studies have shown that engineered DCs effectively deliver antigens and= or cytokines of interest to modulate the type and intensity of immune response (Condon et al, 1996;Figdor et al, 2004;Moll, 2004;Iwashita et al, 2005).…”

“…To date, several viral vector systems including adenovirus, lentivirus, vaccinia virus, as well as herpes simplex virus (HSV)-derived amplicons have been tested for gene delivery to DCs (Takayama et al, 1999;Willis et al, 2001;Di Nicola et al, 2003;Koya et al, 2003Koya et al, , 2004Timares et al, 2004;He et al, 2005). The HSV amplicon possesses a number of advantages over other gene delivery platforms.…”