Abstract
Although many different protocols for transfection of lymphoid cell lines exist, successful DNA transfer into primary B lymphocytes has, to date, not been demonstrated. We now describe a simple method for gene transfer into highly purified normal and malignant B lymphocytes by electroporation. Using a powerful expression vector containing two copies of the cytomegalovirus (CMV) immediate early enhancer linked to the human T cell lymphotropic virus I (HTLV I) promoter, we could demonstrate transfected gene expression in both high density small 'resting' B cells and in low density 'activated' B cells. Successful transfection was detected by expression of chloramphenicol acetyl transferase and by immunofluorescence. The neoplastic cells of B cell chronic lymphocytic leukemia could also be transfected with an efficiency of 5-10%, but only after preactivation. This method of transfection will permit analysis of the contribution of individual genes and their products to normal and malignant B cell growth and differentiation.
Original language | English (US) |
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Pages (from-to) | 77-85 |
Number of pages | 9 |
Journal | Journal of Immunological Methods |
Volume | 133 |
Issue number | 1 |
DOIs | |
State | Published - Oct 4 1990 |
Keywords
- B chronic lymphocytic leukemia
- B lymphocyte
- Chloramphenicol acetyltransferase assay
- Immunofluorescence
- Transfection
ASJC Scopus subject areas
- Immunology
- Biotechnology