GSBS Student Publications

Title

Expression of a human coxsackie/adenovirus receptor transgene permits adenovirus infection of primary lymphocytes

UMMS Affiliation

Graduate School of Biomedical Sciences; Department of Molecular Genetics and Microbiology

Date

10-18-2000

Document Type

Article

Medical Subject Headings

Adenoviridae; Adenoviridae Infections; Animals; B-Lymphocyte Subsets; Cells, Cultured; Crosses, Genetic; Enterovirus; Gene Expression Regulation; Genes, Reporter; Genetic Vectors; Humans; Lymphocyte Activation; Lymphocyte Subsets; Mice; Mice, Inbred C57BL; Mice, Transgenic; Microinjections; Plasmids; Promoter Regions (Genetics); Receptors, Virus; T-Lymphocyte Subsets; Transgenes

Disciplines

Life Sciences | Medicine and Health Sciences

Abstract

Replication-defective adenoviruses are effective vehicles for gene transfer, both for the repair of defective genes and for studies of gene function in primary cells. Many cell types, including lymphocytes, are refractory to adenovirus infection because they lack the Coxsackie/adenovirus receptor (CAR) needed for virus attachment. To extend the advantages of adenovirus-mediated gene transfer to primary lymphoid populations and other cell types lacking endogenous CAR, we produced a mouse that expresses human (h) CAR as a transgene under control of a murine MHC class I promoter. hCAR protein is expressed on T and B lymphocytes from a variety of organs (spleen, lymph node, bone marrow, thymus, and peritoneum). These lymphocytes are susceptible to adenovirus infection, as demonstrated by reporter green fluorescent protein gene expression, with the fraction of expressing cells as high as 70%. Some lymphocyte subpopulations required stimulation subsequent to adenovirus infection for reporter expression. This activation requirement is a restriction imposed by the promoter used in the adenovirus construct. In subpopulations requiring activation, the elongation factor 1 promoter was far superior to a hCMV promoter for directing green fluorescent protein production. We also find that hCAR mRNA is produced in nonlymphoid tissues from all founder lines, including tissues that do not express endogenous murine CAR, suggesting the opportunity for effecting gene delivery to and testing gene function in a wide variety of primary cell types previously resistant to gene transfer.

Rights and Permissions

Citation: J Immunol. 2000 Oct 1;165(7):4112-9.

Related Resources

Link to Article in PubMed

Journal Title

Journal of immunology (Baltimore, Md. : 1950)

PubMed ID

11034423