title: Coxsackie and adenovirus receptor (CAR)-dependent and major histocompatibility complex (MHC) class I-independent uptake of recombinant adenoviruses into human tumour cells.
abstract: The role of two receptors, previously proposed to mediate the entry of adenoviruses into human cells, the coxsackie and adenovirus receptor (CAR) and the major histocompatibility complex (MHC) class I heavy chain has been investigated. The expression of MHC class I in many tumours is reduced or absent, therefore if this were a means by which adenoviruses gained entry into cells, it would have important implications for their application in cancer treatment. In order to determine if MHC class I heavy chain is involved in adenovirus type 5 (Ad5) uptake, the binding of recombinant Ad5 fibre knob domain (which mediates viral attachment) to human cell lines that had greatly different levels of surface MHC class I was studied. We also created derivatives of a non-permissive Chinese hamster ovary (CHO) cell line that expressed human class I (HLA-A2) and found that these cells did not bind fibre or take up virus. In addition, the extracellular domain of CAR was expressed in E. coli and used to generate a polyclonal anti-CAR antibody. This antibody blocked both 125I labelled fibre knob binding and virus uptake. Thus CAR, and not MHC class I, is a receptor for human adenoviruses in cultured tumour cells. Tissue CAR levels may therefore be an important factor in the efficiency of adenovirus-mediated gene therapy.
authors.affiliation: School of Biochemistry and Molecular Biology, University of Leeds, Leeds LS2 9JT, UK.
authors.forename: D
authors.initials: D
authors.lastname: McDonald
authors.forename: L
authors.initials: L
authors.lastname: Stockwin
authors.forename: T
authors.initials: T
authors.lastname: Matzow
authors.forename: M E
authors.initials: ME
authors.lastname: Blair Zajdel
authors.forename: G E
authors.initials: GE
authors.lastname: Blair
mesh_terms: D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D016466:CHO Cells / Q000276:immunology / Q000378:metabolism; D002213:Capsid / Q000378:metabolism; D036022:Capsid Proteins; D062727:Coxsackie and Adenovirus Receptor-Like Membrane Protein; D006224:Cricetinae; D005434:Flow Cytometry; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000378:metabolism*; D006367:HeLa Cells / Q000276:immunology / Q000378:metabolism; D015395:Histocompatibility Antigens Class I / Q000378:metabolism*; D006801:Humans; D018613:Microscopy, Confocal; D011991:Receptors, Virus; D014407:Tumor Cells, Cultured; D016662:Virus Integration
chemical_list: C568771:CLMP protein, human; D036022:Capsid Proteins; D062727:Coxsackie and Adenovirus Receptor-Like Membrane Protein; D015395:Histocompatibility Antigens Class I; D011991:Receptors, Virus; C018315:hexon capsid protein, Adenovirus