Adenoviral-mediated gene transfer in lymphocytes.
Although adenovirus can infect a wide range of cell types, lymphocytes are not generally susceptible to adenovirus infection, in part because of the absence of the expression of the cellular receptor for the adenoviral fiber protein. The cellular receptor for adenovirus and coxsackievirus (CAR) recently was cloned and shown to mediate adenoviral entry by interaction with the viral fiber protein. We show that the ectopic expression of CAR in various lymphocyte cell lines, which are almost completely resistant to adenovirus infection, is sufficient to facilitate the efficient transduction of these cells by recombinant adenoviruses. Furthermore, this property of CAR does not require its cytoplasmic domain, consistent with the idea that CAR primarily serves as a high affinity binding site for the adenoviral fiber protein, and that viral entry is mediated by interaction of the viral penton base proteins with cellular integrins. As a demonstration of their functional utility, we used CAR-expressing lymphocytes transduced with an adenovirus expressing Fas ligand to efficiently kill Fas receptor-expressing tumor cells. The ability to efficiently manipulate gene expression in lymphocyte cells by using adenovirus vectors should facilitate the functional characterization of pathways affecting lymphocyte physiology.
Leon
R P
RP
Department of Biochemistry and Molecular Genetics, University of Colorado Cancer Center, University of Colorado Health Sciences Center, 4200 East Ninth Avenue, Denver, CO 80262, USA.
Hedlund
T
T
Meech
S J
SJ
Li
S
S
Schaack
J
J
Hunger
S P
SP
Duke
R C
RC
DeGregori
J
J
D000256:Adenoviridae / Q000235:genetics* / Q000502:physiology; D000818:Animals; D002460:Cell Line; D062727:Coxsackie and Adenovirus Receptor-Like Membrane Protein; D053222:Fas Ligand Protein; D005822:Genetic Vectors; D006801:Humans; D008214:Lymphocytes / Q000502:physiology*; D008223:Lymphoma; D008562:Membrane Glycoproteins / Q000096:biosynthesis / Q000235:genetics; D051379:Mice; D011991:Receptors, Virus / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D013945:Thymoma; D013953:Thymus Neoplasms; D014162:Transfection / Q000379:methods*; D014407:Tumor Cells, Cultured; D019014:fas Receptor / Q000502:physiology
C568771:CLMP protein, human; C568772:CLMP protein, mouse; D062727:Coxsackie and Adenovirus Receptor-Like Membrane Protein; C505661:FASLG protein, human; D053222:Fas Ligand Protein; C505658:Fasl protein, mouse; D008562:Membrane Glycoproteins; D011991:Receptors, Virus; D011993:Recombinant Fusion Proteins; D019014:fas Receptor