Document 0702
 DOCN  M9650702
 TI    Transduction of rIL-2 expanded CD4+ and CD8+ ovarian TIL-derived T cell
       lines with the G1Na (neor) replication-deficient retroviral vector.
 DT    9605
 AU    Nash MA; Platsoucas CD; Wong BY; Wong PM; Cottler-Fox M; Otto E;
       Freedman RS; Department of Gynecologic Oncology, University of Texas
       M.D.; Anderson Cancer Center, Houston 77030, USA.
 SO    Hum Gene Ther. 1995 Nov;6(11):1379-89. Unique Identifier : AIDSLINE
       MED/96139252
 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.
 DE    Animal  Base Sequence  Cell Line  Cytotoxicity, Immunologic
       CD4-Positive T-Lymphocytes/CYTOLOGY/*IMMUNOLOGY  CD8-Positive
       T-Lymphocytes/CYTOLOGY/*IMMUNOLOGY  DNA Primers  Female  Gene Therapy
       *Genetic Vectors  Human  Interleukin-2/IMMUNOLOGY  Lymphocyte
       Transformation  Lymphocytes, Tumor-Infiltrating/CYTOLOGY/*IMMUNOLOGY
       Mice  Molecular Sequence Data  Ovarian
       Neoplasms/*IMMUNOLOGY/PATHOLOGY/THERAPY  Phosphotransferases (Alcohol
       Group Acceptor)/*GENETICS  Polymerase Chain Reaction
       Retroviridae/*GENETICS/PHYSIOLOGY  Support, Non-U.S. Gov't  Support,
       U.S. Gov't, P.H.S.  *Transformation, Genetic  Tumor Cells, Cultured  3T3
       Cells  JOURNAL ARTICLE

       SOURCE: National Library of Medicine.  NOTICE: This material may be
       protected by Copyright Law (Title 17, U.S.Code).