Gene transfer into humans--immunotherapy of patients with advanced melanoma, using tumor-infiltrating lymphocytes modified by retroviral gene transduction.
METHODS
Treatment with tumor-infiltrating lymphocytes (TIL) plus interleukin-2 can mediate the regression of metastatic melanoma in approximately half of patients. To optimize this treatment approach and define the in vivo distribution and survival of TIL, we used retroviral-mediated gene transduction to introduce the gene coding for resistance to neomycin into human TIL before their infusion into patients--thus using the new gene as a marker for the infused cells.


RESULTS
Five patients received the gene-modified TIL. All the patients tolerated the treatment well, and no side effects due to the gene transduction were noted. The presence and expression of the neomycin-resistance gene were demonstrated in TIL from all the patients with Southern blot analysis and enzymatic assay for the neomycin phosphotransferase coded by the bacterial gene. Cells from four of the five patients grew successfully in high concentrations of G418, a neomycin analogue otherwise toxic to eukaryotic cells. With polymerase-chain-reaction analysis, gene-modified cells were consistently found in the circulation of all five patients for three weeks and for as long as two months in two patients. Cells were recovered from tumor deposits as much as 64 days after cell administration. The procedure was safe according to all criteria, including the absence of infections virus in TIL and in the patients.


CONCLUSIONS
These studies demonstrate the feasibility and safety of using retroviral gene transduction for human gene therapy and have implications for the design of TIL with improved antitumor potency, as well as for the possible use of lymphocytes for the gene therapy of other diseases.
Rosenberg
S A
SA
Division of Cancer Treatment, National Cancer Institute, Bethesda, MD 20892.
Aebersold
P
P
Cornetta
K
K
Kasid
A
A
Morgan
R A
RA
Moen
R
R
Karson
E M
EM
Lotze
M T
MT
Yang
J C
JC
Topalian
S L
SL
D000328:Adult; D004351:Drug Resistance; D005069:Evaluation Studies as Topic; D005260:Female; D005814:Genes, Viral; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D006801:Humans; D007167:Immunotherapy / Q000379:methods*; D007376:Interleukin-2 / Q000008:administration & dosage / Q000627:therapeutic use; D008214:Lymphocytes / Q000187:drug effects / Q000276:immunology*; D008297:Male; D008545:Melanoma / Q000628:therapy*; D008875:Middle Aged; D009355:Neomycin / Q000494:pharmacology; D016133:Polymerase Chain Reaction; D012190:Retroviridae / Q000235:genetics*; D012449:Safety; D014162:Transfection
D007376:Interleukin-2; D009355:Neomycin