Federal Register / Vol. 45, No. 182 / Wednesday, September 17, 1980 / Notices 
61877 
Bn abscess), with subsequent monitoring 
for breakdown of immune tolerance." 
This subject stimulated the most 
discussion during the final plenary 
session of the Workshop. The sentiment 
was to acquire the data as a matter of 
scientific interest rather than to answer 
the risk assessment question. An RFP is 
being developed to solicit applications 
from interested investigators. 
Immunologists from the NIAID staff will 
design the experimental protocol for the 
workscope and other specifications of 
the study which is most likely to be an 
animal model system employing insulin. 
The matter however is still in the 
formative stage and proposers will have 
considerable leeway in outlinging plans 
to answer the primary questions. 
b. Additional information was 
requested on the handling and 
absorption of polypeptides in normal 
and pathologic colon and the potential 
effects of synthetic peptides on the 
bowel itself. 
We have confirmed that this is an 
issue about which there are little direct 
data. The Workshop participants opined 
that although their own calculations 
revealed that only extremely small 
amounts of hormones would be 
produced under the most ideal 
conditions, data should still be sought 
for potential future needs. The issue of 
the potential effects of synthetic 
peptides on the bowel itself is different 
from that of hormones and that 
discussion was centered on interferon 
and its direct effect on cells. 
Although this will be a difficult study 
to initiate because of the technical 
difficulties as well as the process for 
approval of research in human subjects, 
NIAID is considering solicitation of 
grant proposals through an RFA. The 
study will have as it objective a 
determination of the fate of peptide 
hormones when deposited in the distal 
small intestine and large intestine of 
humans. These sites are relevant to the 
production of hormones by recombinant 
DNA technology because they represent 
the regions of Colonization by E. coli. 
c. It was suggested that additional 
information is needed on the potential 
transfer of plasmids to anaerobic 
bacteria from E coli. This 
recommendations differs from the 
findings of the ad hoc group convened in 
August 1979 which RAC reviewed at the 
March 1980 meeting. That expert Group 
noted that by studing the epidemiology 
of plasmids the C'njugafrve plasmids of 
E. coli are not fo nd within the 
anaerobic flora cf the gut. The uniform 
use of non-conjugative plasmids in 
rDNA research further reduces the 
likelihood of transfer. 
It is likely that the requested data will 
be forthcoming from NIH (NIAID) 
supported studies on plasmids through 
the regular grant-supported research 
programs and that new initiatives are 
not needed. In fact, the NIAID Advisory 
Council identified a grant application for 
selective payment that addressed some 
aspects of this issue; this action will 
initiate the acquistion of the data at an 
early date. There is no plan to develop a 
RFA at this time but NIAID staff will 
continue to identify all appropriate 
Incoming applications as potential 
sources of data. 
d. It was recommended that NIH 
communicate with the Center for 
Disease Control (CDC) about possible 
types of health surveillance for workers 
using recombinant DNA. 
A CDC representative was present at 
the Pasadena meeting and discussed 
some approaches to this issue. 
Furthermore, representatives of the 
National Institute for Occupational 
Safety and Health and the Occupational 
Safety and Health Administration were 
also present and therefore, are cognizant 
of the Workshop recommendation. All 
of these agencies are members of The 
Industrial Practices Subcommittee of the 
Federal Interagency Advisory 
Committee on Recombinant DNA 
Research which is discussing this issue. 
5. NIH scientists have continued a 
further evaluation of the biological 
activity of polyoma virus DNA cloned in 
E. coli host vector systems. It had been 
reported (Science 203: 883-892, 1979), 
when the Final Plan was initially 
published that polyoma DNA was 
noninfectious when cloned in EK2 
plasmid and bacteriophage host-vector 
systems. 
As a futher step in evaluating the 
biologic activity of the polyoma-plasmid 
and polyoma-lambda recombinant DNA 
host-vector systems, the scientist tested 
the ability of the EK2 systems containing 
polyoma DNA to induce tumors in 
suckling hamsters. 
These animals are highly sensitive to 
tumor induction by polyoma virus and in 
some cases even subgenomic fragments 
of viral DNA can induce tumors in them. 
The results (Science 205; 1140-1142, 
1979) indicated that inoculation of 
suckling hamsters with 2 X 10* live cells 
of E. coli K-12 strain Chi 1776 carrying 
the complete genome of polyoma virus 
in a recombinant plasmid, failed to 
induce tumors in any of 32 recipients. 
Also, lambda phage DNA end particles 
with a monomeric insert of polyoma 
DNA did not induce tumors. Purified 
recombinant plasmid DNA, as well as 
phage particles and DNA containing a 
hcad-to-tail dimer of polyoma DNA, 
showed a low degree of oncogenicity, 
comparable to that of polyoma DNA 
prepared from mouse cells. These 
findings support the previous 
conclusions, based on infectivity assays 
in mice, that propagation of polyoma 
virus DNA as a component of 
recombinant DNA molecules in E. coli 
K-12 reduces its biologic activity many 
orders of magnitude relative to the virus 
itself. 
Currently both the infectivity and 
tumorigenicity experiments are being 
extended. In one, E. coli K-12 (EKl) 
containing recombinant plasmids 
consisting of two copies of polyoma 
virus DNA and one copy of pBR322 
plasmid DNA are being inoculated into 
newborn hamsters and weanling mice. It 
is also planned to test lambdaphage 
lysogens (£. coli K-12 containing one or 
two copies of a lambda bacteriophage- 
polyoma virus DNA recombinant 
integrated into the bacterial 
chromosome) by inoculation into 
appropriate animal model systems. 
Results will be published in the open 
scientific literature and will be 
presented in a subsequent update of the 
Final Plan. 
6. NIH scientists are also determining 
the biological activity of E. coli K-12 
clones carrying DNA copies of an RNA 
tumor virus. These studies involve the 
administration of bacterial preparations 
containing Harvey sarcoma virus DNA 
to hamsters. These studies are still in 
progress and when definitive results 
have been obtained they will be 
published in the open scientific 
literature and will appear in an annual 
update of the final plan. 
7. Although they nad not appeared in 
the previous Final Plan, two additional 
NIAID contract-supported projects are 
peripherally related to the total risk 
assessment activities. 
a. RAC had suggested that a model 
cloned DNA segment be constructed to 
serve as a uniform heterologous DNA 
segment for risk assessment studies. 
This endeavor has been successfully 
completed by an investigator at the 
University of Wisconsin and employed, 
as a source of DNA, the plasmid NR79. 
The EcoRI fragment NR79-R1-G1 
contains genes encoding for 
chloramphenicol, kanamycin and, 
sulfonamide resistance, as well as the 
promoters that govern these resistances. 
NR79-R1-G1 was shown not to have an 
internal EcoRI site and also not to be 
capable of replicating by itself or with 
the help of other plasmids. Experimental 
results indicated that NR79-R1-G1 is not 
capable of autonomous replication and 
does not contain an origin of replication 
that can function in a recombinant 
plasmid. The antibiotic resistances 
encoded for by NR79-R1-C1 were tested 
•O.S. GOVERNMENT PRINTING OFFICE: 1980-0- >91-132/3231, 
[ 220 ] 
