Generating antibodies against the native form of the human prion protein (hPrP) in wild-type animals: A comparison between DNA and protein immunizations

Electroporation-Aided DNA Immunization Generates Polyclonal Antibodies Against the Native Conformation of Human Endothelin B Receptor

“…Our results also suggest that cardiotoxin pretreatment could accelerate the production of specific antibodies during the course of the immunization protocol (probably through to the recruitment of abundant antigenpresenting cells at the site of injection, Costagliola et al, 1998), which could be advantageous for poorly immunogenic proteins. Another observation of technical interest is the crucial importance of the cell boost after DNA injections (prime-boost strategy) for enhancing antibody titers, confirming previous reports (Nagata et al, 2003;Parise et al, 2008), including ours (Tymciu et al, 2004, Alexandrenne et al, 2009, 2010. It may be noted in connection to this that the negative results of our additional control experiments, using animals injected with hET B R-expressing cells yet without any previous DNA injection, exclude the possibility that the generation of antibodies detected 7 days after the cell injection was solely due to this cellular boost itself.…”

“…levels of antibodies against the native hET B R, as revealed by flow cytometry analysis of intact cells. Our results emphasize the value of applying an electric field to tissues, which increases DNA uptake and gene expression (Mir et al, 1999), as we recently demonstrated in a comparative study of different genetic immunization protocols in a different model (Alexandrenne et al, 2009) and as also reported by others (Kaptein et al, 2008). Our results also suggest that cardiotoxin pretreatment could accelerate the production of specific antibodies during the course of the immunization protocol (probably through to the recruitment of abundant antigenpresenting cells at the site of injection, Costagliola et al, 1998), which could be advantageous for poorly immunogenic proteins.…”

“…As far as GPCRs are concerned, a few antibodies have been prepared up to now using genetic immunization, by various groups (Costagliola et al, 1998, Pichurin et al, 2001, Kaptein et al, 2008, Fujimoto et al, 2009 including our own (Tymciu et al, 2002). More recently, we also showed that the efficiency of DNA immunization against low-immunogenic molecules could be greatly enhanced by using vectors encoding T-cell epitopes (Tymciu et al, 2004), or, above all, by coupling DNA injection to electroporation (Alexandrenne et al, 2009). In the present study, we report that electroporation-aided genetic immunization, possibly coupled to cardiotoxin pretreatment, is a simple and very efficient method of raising large amounts of polyclonal antibodies highly specific for hET B R. The recognition of the native conformation of hET B R was assessed by fluorescence-activated cell sorting (FACS) analysis of different stably transfected cell lines and also by a newly developed, rapid, and efficient cell-based enzyme-linked immunosorbent assay (ELISA) performed on unfixed cells in suspension.…”

CT043, a Protective Antigen That Induces a CD4+Th1 Response duringChlamydia trachomatisInfection in Mice and Humans

“…Moreover, the DNA prime/protein boost regimen is known to elicit both cellular and humoral immunity and has been shown to be effective in vaccination experiments against different pathogens in small animals and nonhuman primates (5,26,39). Finally, DNA immunization could allow the elicitation of an immune response against conformational epitopes (1,32,38). Indeed, a previous study aimed at improving the protective property of a MOMP-based vaccine demonstrated that priming the immune response with MOMP-encoding plasmid, followed by boosting with MOMP, provided a higher protective efficacy in the mouse model than immunization with the recombinant MOMP (8).…”

Therapeutic effect of peripheral administration of an anti-prion protein antibody on mice infected with prions

“…Binding of anti-PrP antibodies to PrP C can inhibit the interaction between PrP C and PrP Sc directly or indirectly (2). Antibody may directly block the domain on PrP C that is required for interacting with PrP Sc ; inhibition of PrP Sc formation by Fab or scFv fragment suggests that this inhibition is a direct effect (6,15,(29)(30)(31). Alternatively, retention of the antibody-PrP C complex may reduce the rate of entry of PrP C into the endocytic pathway, which is reportedly required for PrP Sc formation (32,33 This study shows that passive immunization through a peripheral route may be useful for the treatment of prion diseases.…”