intravenous nutritional support by an indwelling catheter in the superior vena 
cava until they achieved normal hematopoietic function and reconstitution of 
all bleed cell lines. At intervals after transplantation, blood and bone 
marrow samples, or tissues obtained at autopsy, were analyzed for vector DNA 
and for both h-ADA and NPT activities. The results of the first six bone 
mar row/ gene transplants -are summarized in Table 1. 
Cocultivation Protocol 
The first two monkeys received bone marrow that had been cocultured with a 
monolayer of S3A producer cells (Table 1). One of these animals (#855) died of 
systemic bacterial infection 30 days post-transplant without evidence of 
recovery of normal hematopoietic function and was not studied. The other 
animal (#10) experienced a rapid recovery of his white blood cells (achieving 
an absolute neutrophil count of 1000/^In , by day 20), but never achieved a 
normal platelet count throughout the post-transplant period (Figure 3) . Animal 
♦10 was sacrificed at day 69 and the tissues were analyzed for evidence of gene 
transfer. A * *P-labelled-neo gene probe was used for Southern blot analysis 
of DNA f ran peripheral blood mononuclear cells (FBMC), bone marrow and spleen 
following digestion with Kpn I restriction endonuclease. As shewn in Figure 4, 
a 4.9 kb Kpn I band, representing the known unit length fragment characteristic 
of the intact SAX vector (see Fig. 1), was detected in blood and marrow but not 
spleen. This band was also present in the plasmid control lame, but was absent 
in the lane containing ENA from the FBMC of a non-inf ected monkey. Analysis of 
the blot suggests that the equivalent of a single copy of the vector sequence 
was present in 5-10% of blood and marrow cells, since these bands are less than 
1/10 the intensity of the plasmid control which contained a quantity of ENA 
equivalent to approximately one copy/cell. Similar results were obtained 
probing the same blot with a * 'P-labelled-h-ADA gene. Bcwever, in addition to 
Recombinant DNA Research, Volume 12 [285] 
