Transient transfection of primary T helper cells by particle-mediated gene transfer.
The study of the molecular basis of normal CD4+ T cell function, such as the control of commitment to the TH1 or TH2 phenotypes has been difficult due to the resistance of these cells to transfection by conventional methods. We used antibodies specific to T cell surface molecules to immobilize these cells and optimized conditions for transiently transfecting them by means of particle-mediated gene transfer. Using this technique, a construct encompassing - 577 to +1 of the IL-4 promoter allowed transcription of a luciferase reporter gene in recently-differentiated TH2 cells stimulated by anti-CD3, consistent with regulation of endogenous IL-4 gene expression.
Huang
H
H
Laboratory of Immunology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892-1892, USA.
Pannetier
C
C
Hu-Li
J
J
Paul
W E
WE
D000818:Animals; D000918:Antibody Specificity; D015870:Gene Expression; D015847:Interleukin-4 / Q000235:genetics; D008156:Luciferases / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D008213:Lymphocyte Activation / Q000502:physiology; D051379:Mice; D008815:Mice, Inbred Strains; D008822:Mice, Transgenic; D011401:Promoter Regions, Genetic; D006377:T-Lymphocytes, Helper-Inducer / Q000378:metabolism / Q000502:physiology*; D018418:Th2 Cells / Q000378:metabolism / Q000502:physiology; D014162:Transfection / Q000379:methods*
D015847:Interleukin-4; D008156:Luciferases