Gene therapy targeting peripheral blood CD34+ hematopoietic stem cells of HIV-infected individuals

Gene therapy is a promising treatment modality for acquired immunodeficiency syndrome (AIDS), Autologous transplantation with genetically altered pluripotent hematopoietic stem cells encoding anti-human immunodeficiency virus (HIV) genes could in theory completely and permanently reconstitute all blood lineages and immune functions with cells resistant to HIV, Recent studies showed that CD34(+) stem cell can be mobilized in HIV-infected individuals after granulocyte colony-stimulating factor (G-CSF) administration without major side effects or increase of viral load, In this study, peripheral blood CD34(+) cells of five HIV-infected individuals were mobilized with G-CSF and after leukapheresis and enrichment, subjected to retroviral transduction with genes encoding anti-HIV ribozyme-decoy fusion molecules, These cells were tested for the ability to give rise to progeny cells, for retroviral transduction efficiency, and for expression of the transgene, CD34(+)-derived macrophage-like cells were also challenged with HIV. Results showed that CD34(+) cells from HIV-infected individuals gave rise to similar numbers of progeny colonies as cells from healthy donors, The transduction efficiency of these cells varied from 68.8 to 100% as assessed by DNA polymerase chain reaction (PCR) of the transgene in individual colonies, CD34(+)-derived macrophages expressed anti-HIV genes and displayed a substantial and sustained inhibition of HIV replication as compared to untransduced cells, Furthermore, we showed that after thawing, cryopreserved CD34(+) cells from these individuals have survival, proliferation, and transduction parameters comparable to fresh cells, Thus, CD34(+) cells from HIV-infected patients can be stored for further genetic manipulations with improved vectors or anti-HIV genes as they become available.