Introduction of a CD40L genomic fragment via a human artificial chromosome vector permits cell-type-specific gene expression and induces immunoglobulin secretion.
Gene therapy using cDNA driven by an exogenous promoter is not suited for genetic disorders that require intrinsic expression of a transgene, such as hyperimmunoglobulin (Ig)M syndrome (HIGM), which is caused by mutations in the CD40L gene. The human artificial chromosome (HAC) vector has the potential to solve this problem, because it can be used to transfer large genomic fragments containing their own regulatory elements. In this study, we examined whether introduction of a genomic fragment of CD40L via the HAC vector permits intrinsic expression of the transgene and has an effect on immunoglobulin secretion. We constructed an HAC vector carrying the mouse CD40L genomic fragment (mCD40L-HAC) in Chinese hamster ovary (CHO) cells and transferred the mCD40L-HAC vector into a human CD4-positive active T-cell line (Jurkat) and a human myeloid cell line (U937) via microcell-mediated chromosome transfer (MMCT). The mCD40L-HAC vector permits mCD40L expression in human active T cells but not in human myeloid cells. The mCD40L-HAC also functions to stimulate mouse B cells derived from CD40L(-/-) mice, inducing secretion of IgG. This study may be an initial step toward the therapeutic application of HAC vectors for intrinsic expression of genes, a potential new direction for genome-based gene therapy.
Yamada
Hidetoshi
H
Division of Human Genome Science, Department of Molecular and Cellular Biology, Faculty of Medicine, Tottori University, 86, Nishi-cho, Yonago, 683-8503, Japan.|Division of Molecular Genetics and Biofunction, Department of Biomedical Science, Graduate School of Medicine, Tottori University, 86, Nishi-cho, Yonago, 683-8503, Japan.
Li
Yanze C
YC
Division of Human Genome Science, Department of Molecular and Cellular Biology, Faculty of Medicine, Tottori University, 86, Nishi-cho, Yonago, 683-8503, Japan.
Nishikawa
Mitsuo
M
Division of Molecular Genetics and Biofunction, Department of Biomedical Science, Graduate School of Medicine, Tottori University, 86, Nishi-cho, Yonago, 683-8503, Japan.
Oshimura
Mitsuo
M
Division of Molecular Genetics and Biofunction, Department of Biomedical Science, Graduate School of Medicine, Tottori University, 86, Nishi-cho, Yonago, 683-8503, Japan.
Inoue
Toshiaki
T
Division of Human Genome Science, Department of Molecular and Cellular Biology, Faculty of Medicine, Tottori University, 86, Nishi-cho, Yonago, 683-8503, Japan. sour@grape.med.tottori-u.ac.jp.
D000818:Animals; D023201:CD40 Ligand / Q000235:genetics* / Q000502:physiology; D016466:CHO Cells; D002478:Cells, Cultured; D022222:Chromosomes, Artificial, Human / Q000235:genetics*; D006224:Cricetinae; D003412:Cricetulus; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D015894:Genome, Human / Q000235:genetics*; D006801:Humans; D006942:Hypergammaglobulinemia / Q000235:genetics / Q000378:metabolism; D007136:Immunoglobulins / Q000096:biosynthesis*; D019169:Jurkat Cells; D051379:Mice; D018345:Mice, Knockout; D020298:U937 Cells
D007136:Immunoglobulins; D023201:CD40 Ligand