Transfection and gene expression in normal and malignant primary B lymphocytes

Michael Buschle, Malcolm K. Brenner, Irvin S.Y. Chen, Hans G. Drexler, Suzanne M. Gignac, Cliona M. Rooney

Research output: Contribution to journalArticlepeer-review

17 Scopus citations

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 languageEnglish (US)
Pages (from-to)77-85
Number of pages9
JournalJournal of Immunological Methods
Volume133
Issue number1
DOIs
StatePublished - Oct 4 1990

Keywords

  • B chronic lymphocytic leukemia
  • B lymphocyte
  • Chloramphenicol acetyltransferase assay
  • Immunofluorescence
  • Transfection

ASJC Scopus subject areas

  • Immunology
  • Biotechnology

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