Expression of the chimeric IgE gene in cell culture and in various mouse tissues.
Transient expression of recombinant gene constructs is now more widely used in gene therapy as well as in DNA vaccination. In this study, the ability of one and the same genetic construct to drive gene expression both in cell culture and in tissues of the whole organism was demonstrated. Chinese hamster ovarian cells (CHO) were transfected in vitro with plasmids bearing the genes for chimeric IgE (mouse/human) antibodies under control of the human cytomegalovirus (hCMV) promoter. Secretion of recombinant IgE antibodies by transfected cells reached 60% of the intracellular concentration of antibodies. The same gene constructs were introduced into various mouse tissues using ballistic transfection in vivo. The IgE content in blood after transfection of cartilage was found to be several times lower than after transfection of the liver, spleen, or foot pad. At the same time, the content of antibodies to the xenogenous determinants of IgE was essentially independent of the tissue type. These data can be employed in selecting conditions for genetic immunization and gene therapy.
Rodin
D V
DV
Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow 117984, Russia. rodin@imb.ac.ru
Rad'ko
B V
BV
Kolesnikov
V A
VA
Polanovsky
O L
OL
Deyev
S M
SM
D000818:Animals; D000906:Antibodies / Q000378:metabolism; D019470:Biolistics; D016466:CHO Cells; D002356:Cartilage / Q000378:metabolism; D006224:Cricetinae; D003412:Cricetulus; D004797:Enzyme-Linked Immunosorbent Assay; D019084:Fluorescent Antibody Technique, Indirect; D005786:Gene Expression Regulation; D006801:Humans; D007073:Immunoglobulin E / Q000097:blood / Q000235:genetics* / Q000378:metabolism*; D008099:Liver / Q000378:metabolism*; D051379:Mice; D018613:Microscopy, Confocal; D010957:Plasmids; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000097:blood / Q000378:metabolism; D013154:Spleen / Q000378:metabolism*; D014162:Transfection
D000906:Antibodies; D011994:Recombinant Proteins; D007073:Immunoglobulin E