Enhanced interleukin-2 production in human tumor-infiltrating lymphocytes engineered by 3'-truncated interleukin-2 gene.
Tumor-infiltrating lymphocytes (TILs), T lymphocytes associated with solid tumors that can be grown with interleukin (IL)-2 in vitro, preferentially accumulate at tumor sites after adoptive transfer. Therefore, TILs can be considered for use as cellular vehicles in gene therapy. We transduced melanoma TILs with the IL-2 gene and clarified functional characteristics of the TIL transductants. TILs transduced with 3'-end-truncated IL-2 gene (480 bp) produced high amounts of IL-2 detected in supernatants when compared to TILs transduced with the native IL-2 gene containing 3'-end adenine-thymidine (AT)-rich sequences (650 bp). The level of IL-2 in supernatants was higher with the addition of anti-Tac antibody (Ab) to block the consumption of IL-2 by the TILs. These TILs could proliferate autonomously in the absence of exogenous IL-2, and the proliferation of TILs could be completely blocked by anti-IL-2 Ab or anti-IL-2 receptor Ab. Thus TILs transduced with IL-2 gene can proliferate through the autocrine loop. However, the expression of IL-2 from TILs transduced with the IL-2 gene was downregulated after 2 to 3 weeks of G418 selection. Our study indicates the feasibility of transduction and expression of a truncated 480-bp IL-2 gene into TILs and the possibility of employing adoptive immunotherapy protocols using TILs modified with this IL-2 gene.
Yamaue
H
H
Laboratory of Tumor Immunology and Biology, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892.
Kashmiri
S V
SV
De Filippi
R
R
Nieroda
C
C
Yannelli
J R
JR
Tsang
K Y
KY
Schlom
J
J
D001483:Base Sequence; D002455:Cell Division; D003602:Cytotoxicity, Immunologic; D004352:Drug Resistance, Microbial; D005434:Flow Cytometry; D006801:Humans; D007376:Interleukin-2 / Q000235:genetics* / Q000378:metabolism* / Q000494:pharmacology; D016246:Lymphocytes, Tumor-Infiltrating / Q000378:metabolism* / Q000502:physiology*; D008545:Melanoma / Q000235:genetics / Q000473:pathology / Q000628:therapy; D008969:Molecular Sequence Data; D010641:Phenotype; D013997:Time Factors; D014161:Transduction, Genetic; D014407:Tumor Cells, Cultured
D007376:Interleukin-2