Genetic modification of primary chronic lymphocytic leukemia cells with a lentivirus expressing CD38

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Haematologica, Vol 95, Issue 3, 514-517 doi:10.3324/haematol.2009.014381

Genetic modification of primary chronic lymphocytic leukemia cells with a

lentivirus expressing CD38

ce Pearce1, Liam 1, Thet Thet Lin2, Saman Hewamana3, R.

s1, Silvia Deaglio4, Clare Rowntree3, Fegan3,

Pepper2, Brennan1

1 Department of Infection, Immunity and Biochemistry, School of Medicine,

Cardiff University, Cardiff, UK;

2 Department of Haematology, School of Medicine, Cardiff University, Cardiff,

UK;

3 Department of Haematology, University Hospital of Wales, Cardiff, UK and

4 Department of Genetics, Biology and Biochemistry, University of Torino Medical

School & Research Center for Experimental Medicine (CeRMS) Torino, Italy

Correspondence: Brennan, Henry Wellcome Building, School of Medicine,

Cardiff University, Heath Park Cardiff, CF14 4XN, UK. E-mail:

BrennanP@...

Studies of the role of individual genes in chronic lymphocytic leukemia (CLL)

have been hampered by the inability to consistently transfect primary tumor

cells. Here, we describe a highly efficient method of genetically modifying

primary CLL cells using a VSVG pseudotyped lentiviral vector. We transduced CD38

negative CLL cells with a lentiviral vector encoding CD38 which caused increased

surface CD38 expression in all the samples tested (n=17) with no evidence of

plasmacytoid differentiation. The mean percentage of positive cells expressing

CD38 was 87%±8.5% and the mean cell viability 74%±17%. This high level of

transduction of all the CLL cell samples tested demonstrates the utility of this

technique which should prove applicable for the introduction and analysis of

other genes in these non-dividing cells.

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