Table 9B. Feasibility of short-term In vitro expansion and transduction of 
fresh renal tumor explants with the GM-CSF gene after transport to Somatix. 
Fresh renal tumor explants from 5 different patients have been grown in vitro 
over a 4-6 week period and have been transduced with the human GM-CSF gene. 
All 5 patient samples have been grown and transduced at Somatix. The 
transduced cells were assayed by ELISA for GM-CSF secretion in vitro. The 
ELISA assay has a sensitivity limit of 1.5 picograms per ml. Secretion of an 
active GM-CSF protein was confirmed by bioassay which evaluates proliferation 
of a GM-CSF-dependent cell line (TF-1 cells). Transduction efficiency and GM- 
CSF expression does not appear to be affected by histologic subtype. 
TOMOR CODE GM-CSF EXPRESSION POST- LAST IN VITRO HISTOLOGY 
TRANSDUCTION 
PASSAGE 
(ng/10 6 cells/24 hours) 
RCC 
7.1.91 
24 
P13 
Sarcomatoid 
RCC 
7.9.91 
54.7 
P7 
Clear cell, grade II 
RCC 
5.22.92 
34.32 
P3 
Multicystic, Grade III, 
with oncocytic features 
RCC 
6.29.92 
35.05 
P5 
Clear cell. Grade II 
RCC 
7.20.92 
57.6 
P3 
Clear cell. Grade III 
Table 10. Characterization of the fresh human tumor explants for cytokeritin 
production and MHC class I and II levels. Fresh renal tumor explants from 10 
patients were analyzed just after enzymatic digestion for cytokeritin 
production by flow cytometry. Six of these tumors were also assayed again 
after transduction by histochemical evaluation. The in vitro passage at which 
the cells were analyzed is in parenthesis. The histochemical staining is 
employed post-transduction in order to conserve vaccine cells. MHC class I 
and II levels were evaluated for 4 of the tumors evaluated at Somatix. An 
additional 13 renal cell tumors were also evaluated at JHOC for MHC class I 
and II expression All 13 tumors expressed high levels of MHC class I and 
either low or no MHC class II molecules (data not shown) . All tumors were 
assayed for MHC antigen expression by flow cytometry using f luorocein-labeled 
monoclonal antibodies. 
TUMOR 
CODE 
% OF CELLS 
PRODUCING 
KERITIN 
PRE-DIGESTION 
% OF CELLS 
PRODUCING 
KERITIN 
AT TIME OF 
TRANSDUCTION 
MHC CLASS I/II HISTOLOGY 
LEVELS 
RCC 
7.15.91 
50 
High/none 
RCC 
5.22.92 
95 
High/none 
RCC 
6.29.92 
95 
RCC 
7.20.92 
50 
100 
99V0 
RCC 
8.28.92 
60 
80 
(P2) 
100\/0 
RCC 
9.14.92 
87 
RCC 
9.15.92 
50 
95 
(P4) 
RCC 
9.22.92 
40 
100 
RCC 
9.24.92 
100 
(P3 ) 
RCC 
10.1.92 
40 
100 
(P2) 
Sarcomatoid 
Multlcyatlc, grade II.I with 
oncocytic features 
Clear cell Grade II 
Clear cell Grade III 
Grade II/III 
Granular cell 
Granular cell with 
clear cell features 
Grade I/II 
Granular cell 
Papillary, grade II 
Clear Cell Grade III 
Recombinant DNA Research, Volume 17 
[305] 
