PuSH - Publication Server of Helmholtz Zentrum München: Efficient gene transfer of CD40 ligand into primary B-CLL cells using recombinant adeno- associated virus (rAAV) vectors.

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Wendtner, C.-M. ; Kofler, D.M. ; Theiss, H.D. ; Kurzeder, C. ; Buhmann, R.* ; Schweighofer, C.* ; Perabo, L.* ; Danhauser-Riedl, S.* ; Baumert, J.J. ; Hiddemann, W. ; Hallek, M. ; Buning, H.*

Efficient gene transfer of CD40 ligand into primary B-CLL cells using recombinant adeno- associated virus (rAAV) vectors.

Blood 100, 1655-1661 (2002)

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B cells of chronic lymphocytic leukemia (B-CLL) are resistant to transduction with most currently available vector systems. Using an optimized adenovirus-free packaging system, recombinant adeno-associated virus (rAAV) vectors coding for the enhanced green fluorescent protein (AAV/EGFP) and CD40 ligand (AAV/CD40L) were packaged and highly purified resulting in genomic titers up to 3 × 10¹¹/mL. Cells obtained from 24 patients with B-CLL were infected with AAV/EGFP or AAV/CD40L at a multiplicity of infection (MOI) of 100 resulting in transgene expression in up to 97% of cells as detected by flow cytometry 48 hours after infection. Viral transduction could be specifically blocked by heparin. Transduction with AAV/CD40L resulted in up-regulation of the costimulatory molecule CD80 not only on infected CLL cells but also on noninfected bystander leukemia B cells, whereas this effect induced specific proliferation of HLA-matched allogeneic T cells. Vaccination strategies for patients with B-CLL using leukemia cells infected ex vivo by rAAV vectors now seems possible in the near future.

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