Human T lymphocytes transduced by lentiviral vectors in the absence of TCR activation maintain an intact immune competence.
Gene transfer into T lymphocytes is currently being tested for the treatment of lymphohematologic disorders. We previously showed that suicide gene transfer into donor lymphocytes infused to treat leukemic relapse after allogeneic hematopoietic stem cell transplantation allowed control of graft-versus-host disease. However, the T-cell receptor (TCR) activation and sustained proliferation required for retroviral vector transduction may impair the half-life and immune competence of transduced cells and reduce graft-versus-leukemia activity. Thus, we tested lentiviral vectors (LVs) and stimulation with cytokines involved in antigen-independent T-cell homeostasis, such as interleukin 7 (IL-7), IL-2, and IL-15. Late-generation LVs transduced efficiently nonproliferating T cells that had progressed from G0 to the G1 phase of the cell cycle on cytokine treatment. Importantly, IL-2 and IL-7, but not IL-15, stimulation preserved physiologic CD4/CD8 and naive-memory ratios in transduced cells with only minor induction of some activation markers. Functional analysis of immune response to cytomegalovirus (CMV) showed that, although CMV-specific T cells were preserved by all conditions of transduction, proliferation and specific killing of autologous cells presenting a CMV epitope were higher for IL-2 and IL-7 than for IL-15. Thus, LV transduction of IL-2 or IL-7 prestimulated cells overcomes the limitations of retroviral vectors and may significantly improve the efficacy of T-cell-based gene therapy.
Cavalieri
Simona
S
Institute for Cancer Research and Treatment, University of Torino Medical School, Cadiolo Torino, Italy.
Cazzaniga
Sabrina
S
Geuna
Massimo
M
Magnani
Zulma
Z
Bordignon
Claudio
C
Naldini
Luigi
L
Bonini
Chiara
C
D000956:Antigens, Viral / Q000276:immunology; D002453:Cell Cycle; D002460:Cell Line; D003587:Cytomegalovirus / Q000276:immunology; D017930:Genes, Reporter; D005822:Genetic Vectors / Q000235:genetics*; D006367:HeLa Cells; D006801:Humans; D007121:Immunocompetence; D007371:Interferon-gamma / Q000096:biosynthesis; D019409:Interleukin-15 / Q000494:pharmacology; D007376:Interleukin-2 / Q000494:pharmacology; D015851:Interleukin-7 / Q000494:pharmacology; D007668:Kidney / Q000166:cytology; D016086:Lentivirus / Q000235:genetics*; D008213:Lymphocyte Activation; D020800:Receptor, Nerve Growth Factor / Q000235:genetics; D011948:Receptors, Antigen, T-Cell / Q000276:immunology; D016176:T-Lymphocyte Subsets / Q000187:drug effects / Q000276:immunology* / Q000821:virology; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology; D014161:Transduction, Genetic
D000956:Antigens, Viral; D019409:Interleukin-15; D007376:Interleukin-2; D015851:Interleukin-7; D020800:Receptor, Nerve Growth Factor; D011948:Receptors, Antigen, T-Cell; D007371:Interferon-gamma