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.
Gervaix
A
A
Department of Medicine, University of California at San Diego, La Jolla 92093-0665, USA.
Schwarz
L
L
Law
P
P
Ho
A D
AD
Looney
D
D
Lane
T
T
Wong-Staal
F
F
D000818:Animals; D018952:Antigens, CD34; D002460:Cell Line; D002472:Cell Transformation, Viral; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D016179:Granulocyte Colony-Stimulating Factor / Q000008:administration & dosage; D015658:HIV Infections / Q000097:blood* / Q000628:therapy; D015497:HIV-1 / Q000472:pathogenicity; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D006801:Humans; D007937:Leukapheresis; D007958:Leukocyte Count; D007963:Leukocytes, Mononuclear / Q000166:cytology / Q000378:metabolism; D008264:Macrophages / Q000166:cytology / Q000378:metabolism; D051379:Mice; D008979:Moloney murine leukemia virus; D009000:Monocytes / Q000166:cytology / Q000378:metabolism; D016337:RNA, Catalytic / Q000096:biosynthesis* / Q000235:genetics; D014407:Tumor Cells, Cultured
D018952:Antigens, CD34; D016337:RNA, Catalytic; D016179:Granulocyte Colony-Stimulating Factor