PMID- 8573611
OWN - NLM
STAT- MEDLINE
DCOM- 19960314
LR  - 20121115
IS  - 1043-0342 (Print)
IS  - 1043-0342 (Linking)
VI  - 6
IP  - 11
DP  - 1995 Nov
TI  - Transduction of rIL-2 expanded CD4+ and CD8+ ovarian TIL-derived T cell lines 
      with the G1Na (neor) replication-deficient retroviral vector.
PG  - 1379-89
AB  - We have expanded ovarian tumor-infiltrating lymphocytes (TIL) in low 
      concentrations of recombinant interleukin-2 (rIL-2) to conduct intraperitoneal 
      adoptive immunotherapy trials in patients with ovarian cancer. We have previously 
      demonstrated that certain T cell lines and clones derived from ovarian TIL 
      exhibit in vitro autologous tumor-specific cytotoxicity and/or cytokine 
      production (interferon-gamma, tumor necrosis factor-alpha) preferentially in 
      response to autologous tumor cells. Studies that utilize a marker gene introduced 
      into the DNA of TIL can provide useful information on specific uptake or 
      localization of TIL at tumor sites and on the survival of TIL in vivo. We have 
      conducted a series of preclinical experiments in which we have successfully 
      transfected TIL with G1Na, which encodes the gene for neomycin phosphotransferase 
      (neoR). NeoR was detected in at least 10% of CD8+ cells (mean = 10.4%) and 
      between 2.5 and 20% of CD4+ TIL (mean = 8.5%). Transduction of ovarian TIL with 
      G1Na caused no substantial changes to the T cell phenotypes or in vitro 
      cytotoxicities against ovarian and hematogenous tumor cell targets, or on the 
      rIL-2 requirements of TIL for growth and proliferation. In addition, the intact 
      G1Na provirus in transduced TIL cells was rescuable by replication-competent 
      retrovirus and was transferred into the genome of NIH-3T3 fibroblasts, which were 
      rendered resistant to G418. An enhanced polymerase chain reaction (PCR) procedure 
      utilizing detection by ethidium bromide staining was developed. The enhanced PCR 
      detected 1 in 100,000 neoR-labeled cells. Furthermore, detection of the G1Na 
      genome in transduced TIL by in situ hybridization with an RNA probe provided 
      evidence for expression of the neoR gene in transduced TIL. Results obtained from 
      these studies suggest that ovarian TIL-derived T cell lines transduced with the 
      neoR gene post infection with the G1Na retroviral vector can be utilized to 
      examine the in vivo trafficking pattern of ovarian TIL-derived T cell lines 
      expanded in low concentrations of rIL-2 and their survival.
FAU - Nash, M A
AU  - Nash MA
AD  - Department of Gynecologic Oncology, University of Texas M.D. Anderson Cancer 
      Center, Houston 77030, USA.
FAU - Platsoucas, C D
AU  - Platsoucas CD
FAU - Wong, B Y
AU  - Wong BY
FAU - Wong, P M
AU  - Wong PM
FAU - Cottler-Fox, M
AU  - Cottler-Fox M
FAU - Otto, E
AU  - Otto E
FAU - Freedman, R S
AU  - Freedman RS
LA  - eng
GR  - CA-52308/CA/NCI NIH HHS/United States
GR  - CA-57884/CA/NCI NIH HHS/United States
GR  - CA-64943-01/CA/NCI NIH HHS/United States
GR  - etc.
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - Hum Gene Ther
JT  - Human gene therapy
JID - 9008950
RN  - 0 (DNA Primers)
RN  - 0 (Interleukin-2)
RN  - EC 2.7.1.- (Phosphotransferases (Alcohol Group Acceptor))
RN  - EC 2.7.1.95 (Kanamycin Kinase)
SB  - IM
MH  - 3T3 Cells
MH  - Animals
MH  - Base Sequence
MH  - CD4-Positive T-Lymphocytes/cytology/*immunology
MH  - CD8-Positive T-Lymphocytes/cytology/*immunology
MH  - Cell Line
MH  - Cytotoxicity, Immunologic
MH  - DNA Primers
MH  - Female
MH  - Genetic Therapy
MH  - *Genetic Vectors
MH  - Humans
MH  - Interleukin-2/immunology
MH  - Kanamycin Kinase
MH  - Lymphocyte Activation
MH  - Lymphocytes, Tumor-Infiltrating/cytology/*immunology
MH  - Mice
MH  - Molecular Sequence Data
MH  - Ovarian Neoplasms/*immunology/pathology/therapy
MH  - Phosphotransferases (Alcohol Group Acceptor)/*genetics
MH  - Polymerase Chain Reaction
MH  - Retroviridae/*genetics/physiology
MH  - *Transformation, Genetic
MH  - Tumor Cells, Cultured
EDAT- 1995/11/01 00:00
MHDA- 1995/11/01 00:01
CRDT- 1995/11/01 00:00
PHST- 1995/11/01 00:00 [pubmed]
PHST- 1995/11/01 00:01 [medline]
PHST- 1995/11/01 00:00 [entrez]
AID - 10.1089/hum.1995.6.11-1379 [doi]
PST - ppublish
SO  - Hum Gene Ther. 1995 Nov;6(11):1379-89. doi: 10.1089/hum.1995.6.11-1379.