Transfection and gene expression in normal and malignant primary B lymphocytes

J Immunol Methods. 1990 Oct 4;133(1):77-85. doi: 10.1016/0022-1759(90)90321-l.


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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Antigens, Differentiation
  • B-Lymphocytes* / enzymology
  • B-Lymphocytes* / immunology
  • CD5 Antigens
  • Cell Separation
  • Chloramphenicol O-Acetyltransferase / genetics
  • Cytomegalovirus / genetics
  • Electric Stimulation
  • Enhancer Elements, Genetic / genetics
  • Fluorescent Antibody Technique
  • Gene Expression*
  • Herpesvirus 4, Human / genetics
  • Human T-lymphotropic virus 1 / genetics
  • Humans
  • Leukemia, Lymphocytic, Chronic, B-Cell / genetics
  • Leukemia, Lymphocytic, Chronic, B-Cell / immunology*
  • Lymphocyte Activation / drug effects
  • Plasmids
  • Tetradecanoylphorbol Acetate / pharmacology
  • Transfection*
  • Tumor Cells, Cultured


  • Antigens, Differentiation
  • CD5 Antigens
  • Chloramphenicol O-Acetyltransferase
  • Tetradecanoylphorbol Acetate