title: Vaccine Design: Replication-Defective Adenovirus Vectors.
abstract: Replication-defective adenovirus (Ad) vectors were initially developed for gene transfer for correction of genetic diseases. Although Ad vectors achieved high levels of transgene product expression in a variety of target cells, expression of therapeutic proteins was found to be transient as vigorous T cell responses directed to components of the vector as well as the transgene product rapidly eliminate Ad vector-transduced cells. This opened the use of Ad vectors as vaccine carriers and by now a multitude of preclinical as well as clinical studies has shown that Ad vectors induce very potent and sustained transgene product-specific T and B cell responses. This chapter provides guidance on developing E1-deleted Ad vectors based on available viral molecular clones. Specifically, it describes methods for cloning, viral rescue and purification as well as quality control studies.
authors.affiliation: The Wistar Institute, Philadelphia, PA, 19104, USA.
authors.forename: Xiangyang
authors.initials: X
authors.lastname: Zhou
authors.affiliation: The Wistar Institute, Philadelphia, PA, 19104, USA.
authors.forename: Zhiquan
authors.initials: Z
authors.lastname: Xiang
authors.affiliation: The Wistar Institute, Philadelphia, PA, 19104, USA. ertl@wistar.org.
authors.forename: Hildegund C J
authors.initials: HCJ
authors.lastname: Ertl
mesh_terms: D000260:Adenoviruses, Human / Q000235:genetics* / Q000502:physiology*; D002498:Centrifugation; D003001:Cloning, Molecular; D005818:Genetic Engineering / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics*; D057809:HEK293 Cells; D006801:Humans; D016133:Polymerase Chain Reaction; D011786:Quality Control; D014162:Transfection; D019076:Transgenes / Q000235:genetics; D014612:Vaccines / Q000235:genetics*; D014779:Virus Replication
chemical_list: D014612:Vaccines
keywords: Adenovirus; Cloning; Vector purification; Vector quality control; Viral molecular clone; Viral rescue