In vivo bioluminescence imaging in preclinical trials of genetic vaccines

摘要: DNA immunization is a rapidly developing vaccine platform for cancer, infectious disease, and allergies. The efficiency of vaccination largely determined by the delivery subsequent expression HIV-1 genes in cells. immunogens are generally administered intramuscular or intradermal injections, followed electroporation to enhance uptake into An intense debate on pros cons different routes still ongoing. A number studies have compared effect methods amount quality DNA-directed immunogen expression, as well magnitude specificity immune response they generate. Several were based post mortem tissues, indirect monitored techniques such vivo imaging reporter co-delivered fused immunogens. aim this work was develop applications immunization. first optimize order increase efficacy immunogenicity vaccines. Furthermore we set out use differences strength type induced (ID) (IM) injection routes, each electroporation. In particular, task determine extent which method influences Th1 Th2 immunogens, represented viral protease (PR) reverse transcriptase (RT) HIV-1, respectively. Our final objective acquired results an attempt model responses silico. BALB/c mice immunized with mixed gene encoding bioluminescent reporter. We used bioluminescence (BLI) tool monitor delivered vivo. By combining readouts form BLI immunoassays able produce parameters that result best outcome terms immunogenicity. Upon optimization conditions exploited one suited providing maximal Here show ID administration significant enhancement both cellular humoral IM. evident regardless nature (Th1 vs. Th2). kinetics loss found correlate antigen-specific production IFN-γ IL-2 could thus be characterize specific against immunogen. Thus, assess non-invasive from reporters. reporters new strategy their start completion experiment. level presence reflects construct, presenting anon-invasive (technique) dynamics individual recipients useful determination study end-points. application technique allows us significantly refine reduce animal experimentation development. LIST OF SCIENTIFIC PAPERS I. Petkov S, Heuts F, Krotova O, Kilpelainen A, Engstrom G, Starodubova E, Isaguliants M. Evaluation using imaging. Hum. Vaccin. Immunother 2013, Oct; 9(10): 2228-2236. II. Kostic L, Agapkina J, Hallengard D, Viklund Latyshev Gelius Dillenbeck T, Karpov V, Gottikh M, Belyakov I, Lukashov Consensus FSU-A integrase variants electroporated induce polyfunctional CD4+ CD8+ T PLoS ONE 201, 8(5): e62720. III. Latanova Expression localization determines but not mice. Manuscript. CONTENTS