| Abstract: |
Crawling dendritic cells (CDCs) and herpes simplex virus-1 (HSV-1) amplicon vectors were utilized in this study: (1) to evaluate whether CDCs can be transduced by HSV-1 amplicon vectors; (2) to assess the effects of HSV-1 infections on structure and functions of CDCs; (3) to assess the capabilities of the transduced CDC to express, process, and present the transgene products; and (4) to induce in vitro and in vivo priming of T cells and B cells. CDC supported amplicon-mediated transgene expression while retaining the ability to perform mixed lymphocyte reaction (MLR) and priming of naive T cells. Then it was tested whether transduced CDC were able to initiate immunity against either the amplicon particle and/or the product encoded by the delivered transgene by injecting groups of mice with transduced CDCs expressing GFP or LacZ. Spleen cells of these mice were stimulated by co-incubation with cells expressing: (1) either one of the transgenes (GFP or LacZ), (2) peptides of β-gal, or (3) peptides of HSV-1 glycoprotein B (gB). Interestingly, no significant cytotoxic T lymphocyte (CTL) activity against the transgenes or against gB was observed. In contrast, mice developed high levels of antibodies against gB and LacZ. Mainly, the findings that CDCs not only express amplicon-delivered transgene, but were able to induce MLR and priming of naïve T cells against the transduced antigen, open up unexpected possibilities and the likelihood to use CDCs as a vehicle for cellular immunization against any transduced antigens. However, these results indicate that HSV-1 amplicon-transduced CDCs induce effective priming and a humoral response, but no strong cell-mediated immune response. |
| Keywords: |
controlled study; nonhuman; flow cytometry; t lymphocyte; t-lymphocytes; animal cell; mouse; animals; mice; cells, cultured; cell structure; infection; gene expression; dendritic cell; green fluorescent protein; animal experiment; herpes simplex; mice, inbred c57bl; b lymphocyte; genetic transduction; genetic vectors; antigen presentation; lymphocyte activation; cytokines; dendritic cells; lymphocyte culture test, mixed; genetic transfection; amplicon; vaccination; cytotoxic t lymphocyte; transgene; beta galactosidase; green fluorescent proteins; cytokine production; genes, reporter; herpesvirus 1, human; cell stimulation; humoral immunity; herpes simplex virus 1; cell activity; antibody; transgenes; immunologic memory; antibody production; mixed lymphocyte reaction; cytopathogenic effect, viral; spleen cell; nonviral gene delivery system; gene transfer techniques; transduction; antigens, viral; virus glycoprotein; cell based gene therapy; herpesvirus vector; humans; priority journal; article; amplicon-hsv-1; cdc priming; cellular vaccine
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