Transient expression of the CD2 cell surface antigen as a sortable marker to monitor high frequency transfection of human primary B cells.
We have used the T cell surface molecule CD2 gene, expressed from the human cytomegalovirus promoter as a reporter to optimize a transfection system for human primary B cells. The CD2-encoding DNA was transfected into cells by electroporation and transient expression was monitored by flow cytometric analysis. By using our optimal electroporation conditions on activated primary B cells, more than 30% of the resulting viable cells expressed CD2 on the cell surface. Moreover, unactivated primary B cells could also be transfected using this system but subsequent expression of CD2 required cellular activation. Magnetic beads or plastic culture bottles coated with anti-CD2 antibodies have been used to selectively purify transfected cells. The high transfection efficiency combined with the ability to specifically purify transfected cells may allow future studies on specific genes transiently expressed in human primary B cells.
Pilon
M
M
Unit for Applied Cell and Molecular Biology, University of Umeå, Sweden.
Gullberg
M
M
Lundgren
E
E
D000945:Antigens, Differentiation, T-Lymphocyte / Q000032:analysis / Q000235:genetics*; D001402:B-Lymphocytes / Q000276:immunology*; D018801:CD2 Antigens; D015500:Chloramphenicol O-Acetyltransferase / Q000032:analysis; D015870:Gene Expression; D006801:Humans; D011971:Receptors, Immunologic / Q000032:analysis / Q000235:genetics*; D014162:Transfection
D000945:Antigens, Differentiation, T-Lymphocyte; D018801:CD2 Antigens; D011971:Receptors, Immunologic; D015500:Chloramphenicol O-Acetyltransferase