Generation of multi-functional antigen-specific human T-cells by lentiviral TCR gene transfer.
T-cell receptor (TCR) gene transfer is an attractive strategy to generate antigen-specific T-cells for adoptive immunotherapy of cancer and chronic viral infection. However, current TCR gene transfer protocols trigger T-cell differentiation into terminally differentiated effector cells, which likely have reduced ability to mediate disease protection in vivo. We have developed a lentiviral gene transfer strategy to generate TCR-transduced human T-cells without promoting T-cell differentiation. We found that a combination of interleukin-15 (IL15) and IL21 facilitated lentiviral TCR gene transfer into non-proliferating T-cells. The transduced T-cells showed redirection of antigen specificity and produced IL2, IFNgamma and TNFalpha in a peptide-dependent manner. A significantly higher proportion of the IL15/IL21-stimulated T-cells were multi-functional and able to simultaneously produce all three cytokines (P<0.01), compared with TCR-transduced T-cells generated by conventional anti-CD3 plus IL2 stimulation, which primarily secreted only one cytokine. Similarly, IL15/IL21 maintained high levels of CD62L and CD28 expression in transduced T-cells, whereas anti-CD3 plus IL2 accelerated the loss of CD62L/CD28 expression. The data demonstrate that the combination of lentiviral TCR gene transfer together with IL15/IL21 stimulation can efficiently redirect the antigen specificity of resting primary human T-cells and generate multi-functional T-cells.
Perro
M
M
Department of Immunology, Division of Infection and Immunity, University College London, Royal Free Hospital, London, UK.
Tsang
J
J
Xue
S-A
SA
Escors
D
D
Cesco-Gaspere
M
M
Pospori
C
C
Gao
L
L
Hart
D
D
Collins
M
M
Stauss
H
H
Morris
E C
EC
D002454:Cell Differentiation; D049109:Cell Proliferation; D000939:Epitopes; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D006801:Humans; D016219:Immunotherapy, Adoptive / Q000379:methods; D018664:Interleukin-12 / Q000235:genetics*; D019409:Interleukin-15 / Q000235:genetics*; D016086:Lentivirus / Q000235:genetics*; D011948:Receptors, Antigen, T-Cell / Q000235:genetics*; D013601:T-Lymphocytes / Q000276:immunology*; D014161:Transduction, Genetic
D000939:Epitopes; D019409:Interleukin-15; D011948:Receptors, Antigen, T-Cell; D018664:Interleukin-12