to be etiologic in these rodent models. We have no reason 
to believe that the malignancies were related to the 
recombinant neomycin-resistance vector, since the tumor 
cells did not contain the marker gene. These events 
emphasize the critical importance of using retroviral 
supernatants that are devoid of competent helper virus in 
all human clinical protocols. ’ . 
Anderson and Rosenberg at NIH have used LNL6 to mark tumor- 
infiltrating lymphocytes (TIL cells) in patients with 
metastatic malignancies and have then followed the fate of 
the TIL cells in vivo after reinfusion. 56 Marked TIL cells 
could be detected in the patients post-infusion, and the 
phenotype of marked cells was unchanged by the procedure. 
No horizontal transmission of LNL6 was detected. Protocols 
are ongoing at St. Jude's Hospital and M.D. Anderson Cancer 
Center using retroviral gene transfer to mark autologous 
marrow grafts. Gene therapy protocols in ADA-deficient SCID, 
melanoma, renal cell cancer, and hyperlipidemia patients are 
ongoing, and no adverse effects resulting from gene transfer 
have yet been reported. A small number of patients with 
severe combined immunodeficiency have received infusions of 
autologous lymphocytes transduced with an adenosine 
deaminase vector, and have shown some evidence for improved 
immune function without adverse ef f ects .XMichael Blaese, 
personal communication) 
Introduction of a Neo* gene: The Neo R gene product, neomycin 
phosphotransferase, phosphorylates the 3' hydroxyl group of 
neomycin and its analogues, thus inactivating the 
antibiotic. It is possible that amikacin is partially 
inavtivated by this enzyme, but other antibiotics in 
clinical usage, including gentamycina and tobramycin, do not 
contain this 3' hydroxyl group and thus are not inactivated 
by the Neo* gene product. 
. 0 PATIENT ELIGIBILITY 
All patients eligible for the original multiple myeloma (92- 
H-57), chronic myelogenous leukemia (92-H-56), and breast 
cancer autologous transplantation protocols will be enrolled 
in this protocol amendment if they meet the criteria 
outlined below. (See Section 4.0 of each of the protocols 
for specific criteria) Patients will sign an additional 
informed consent for the genetic marking amendment. 
Additional criteria include: 
4 . 1 Adequacy of Peripheral Blood Stem Cell Collection : 
(Multiple Myeloma and CML protocols only) The patient 
will undergo apheresis as described in the original 
protocols beginning on the dav that total ! o’^-ocyf $ 
Recombinant DNA Research, Volume 16 
[17] 
