title: I. Poloxamer-formulated plasmid DNA-based human cytomegalovirus vaccine: evaluation of plasmid DNA biodistribution/persistence and integration.
abstract: Preclinical studies were conducted in mice and rabbits to evaluate biodistribution/persistence and potential integration of plasmid DNA (pDNA) after intramuscular administration of a poloxamer-formulated pDNAbased vaccine, VCL-CT01, encoding gB, pp65, and IE1 human cytomegalovirus (hCMV) immunogens. Tissue distribution in mice vaccinated with VCL-CT01 was compared with that in mice vaccinated with a phosphate- buffered saline (PBS)-formulated control pDNA vaccine. Residual pDNA copy number (PCN), in selected tissues collected on days 3, 30, and 60 after vaccination, was measured by quantitative polymerase chain reaction. In VCL-CT01-vaccinated mice and in control pDNA-vaccinated mice, pDNA was below the limit of detection by day 60 in all tissues except the injection site. Clearance of pDNA from the injection site was slower in VCL-CT01-vaccinated mice compared with PBS-pDNA-vaccinated mice. An integration study was conducted in rabbits to determine whether pDNA integration into the genome of the vaccinated animal contributed to pDNA persistence. Residual pDNA in VCL-CT01-injected rabbit muscle collected 60 days after vaccination (geometric mean of 1085 PCN/microg total DNA) was comparable to that observed in VCL-CT01- injected mouse muscle (geometric mean of 1471 PCN/microg total DNA) collected at the same time point. pDNA integration was not detectable by column agarose gel electrophoresis despite the persistence of pDNA at the injection site 60 days after vaccination. Therefore the risk of genomic integration of hCMV pDNA formulated with poloxamer was considered negligible.
authors.affiliation: Vical Incorporated, San Diego, CA 92121, USA. avilalta@vical.com
authors.forename: Adrián
authors.initials: A
authors.lastname: Vilalta
authors.forename: Rohit K
authors.initials: RK
authors.lastname: Mahajan
authors.forename: Jukka
authors.initials: J
authors.lastname: Hartikka
authors.forename: Denis
authors.initials: D
authors.lastname: Rusalov
authors.forename: Terrie
authors.initials: T
authors.lastname: Martin
authors.forename: Vesselina
authors.initials: V
authors.lastname: Bozoukova
authors.forename: Vicky
authors.initials: V
authors.lastname: Leamy
authors.forename: Keith
authors.initials: K
authors.lastname: Hall
authors.forename: Peggy
authors.initials: P
authors.lastname: Lalor
authors.forename: Alain
authors.initials: A
authors.lastname: Rolland
authors.forename: David C
authors.initials: DC
authors.lastname: Kaslow
mesh_terms: D000818:Animals; D003587:Cytomegalovirus / Q000235:genetics / Q000276:immunology; D003586:Cytomegalovirus Infections / Q000235:genetics / Q000276:immunology / Q000517:prevention & control; D022482:Cytomegalovirus Vaccines / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology / Q000493:pharmacokinetics*; D004353:Drug Evaluation, Preclinical; D006801:Humans; D007273:Injections, Intramuscular; D051379:Mice; D020442:Poloxamer / Q000737:chemistry / Q000493:pharmacokinetics*; D011817:Rabbits; D019444:Vaccines, DNA / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology / Q000493:pharmacokinetics*; D014764:Viral Proteins / Q000235:genetics / Q000276:immunology*
chemical_list: D022482:Cytomegalovirus Vaccines; D019444:Vaccines, DNA; D014764:Viral Proteins; D020442:Poloxamer