title: Influence of fiber detargeting on adenovirus-mediated innate and adaptive immune activation.
abstract: The major adenovirus (Ad) capsid proteins hexon, penton, and fiber influence the efficiency and tropism of gene transduction by Ad vectors. Fiber is the high-affinity receptor binding protein that serves to mediate cell attachment in vitro when using coxsackie-adenovirus receptor (CAR)-containing cell lines. This contrasts with transduction efficiency in macrophages or dendritic cells that lack high concentrations of CAR. To determine how fiber influences gene transduction and immune activation in a murine model, we have characterized Ad type 5 (Ad5) vectors with two classes of chimeric fiber, CAR binding and non-CAR binding. In a systemic infection, Ad5 fiber contributes to DNA localization and vector transduction in hepatic tissue. However, the majority of vector localization is due to Ad5 fiber-specific functions distinct from CAR binding. CAR-directed transduction occurs but at a modest level. In contrast to CAR binding vectors, the F7 and F7F41S non-CAR-binding vectors demonstrate a 2-log decrease in hepatic transduction, with a 10-fold decrease in the amount of vector DNA localizing to the hepatic tissue. To characterize the innate response to early infection using fiber chimeric vectors, intrahepatic cytokine and chemokine mRNAs were quantified 5 hours postinfection. Tumor necrosis factor alpha mRNA levels resulting from Ad5 fiber infections were elevated compared to viruses expressing serotype 7 or 41 fiber. Levels of chemokine mRNA (gamma interferon-inducible protein 10, T-cell activation gene 3, and macrophage inflammatory protein 1beta) were 10- to 20-fold higher with CAR binding vectors (Ad5 and F41T) than with non-CAR-binding vectors (F7 and F7F41S). In spite of quantitative differences in vector localization and innate activation, fiber pseudotyping did not significantly change the outcome of anti-Ad adaptive immunity. All vectors were cleared with the same kinetics as wild-type Ad5 vectors, and each induced neutralizing antibody. Although non-CAR-binding vectors were impaired in transduction by nearly 2 orders of magnitude, the level of antitransgene immunity was the same for each of the vectors. Using primary bone marrow-derived macrophages and dendritic cells, we demonstrate that transduction, induction of cytokine/chemokine, and phenotypic maturation of these antigen-presenting cells are independent of fiber content. Our data support a model where fiber-mediated hepatic localization enhances innate responses to virus infection but minimally impacts on adaptive immunity.
authors.affiliation: Weill Medical College of Cornell University, Hearst Research Foundation, Department of Microbiology and Immunology Graduate Program, Box 62, 1300 York Ave., New York, New York 10021, USA.
authors.forename: John W
authors.initials: JW
authors.lastname: Schoggins
authors.forename: Marcelo
authors.initials: M
authors.lastname: Nociari
authors.forename: Nicola
authors.initials: N
authors.lastname: Philpott
authors.forename: Erik
authors.initials: E
authors.lastname: Falck-Pedersen
mesh_terms: D000222:Adaptation, Physiological; D000256:Adenoviridae / Q000235:genetics* / Q000276:immunology* / Q000502:physiology; D000818:Animals; D000956:Antigens, Viral / Q000235:genetics; D036022:Capsid Proteins / Q000235:genetics* / Q000276:immunology*; D002460:Cell Line; D018925:Chemokines / Q000378:metabolism; D062727:Coxsackie and Adenovirus Receptor-Like Membrane Protein; D016207:Cytokines / Q000378:metabolism; D004279:DNA, Viral / Q000235:genetics / Q000378:metabolism; D003713:Dendritic Cells / Q000276:immunology / Q000821:virology; D005260:Female; D005822:Genetic Vectors; D022781:Hepatocytes / Q000276:immunology / Q000821:virology; D006801:Humans; D007113:Immunity, Innate; D008264:Macrophages / Q000276:immunology / Q000821:virology; D051379:Mice; D011991:Receptors, Virus / Q000235:genetics / Q000276:immunology; D014161:Transduction, Genetic
chemical_list: D000956:Antigens, Viral; C568771:CLMP protein, human; C568772:CLMP protein, mouse; D036022:Capsid Proteins; D018925:Chemokines; D062727:Coxsackie and Adenovirus Receptor-Like Membrane Protein; D016207:Cytokines; D004279:DNA, Viral; D011991:Receptors, Virus; C018315:hexon capsid protein, Adenovirus; C043322:penton protein, adenovirus