55104 
Federal Register / Vul. 47, No. 235 / Tuesday, December 7, 1902 / Notices 
DEPARTMENT OF HEALTH AND 
HUMAN SERVICES 
National Institutes of Health 
Program To Assess the Risks of 
Recombinant DNA Research; 
Proposed Second Annual Update 
agency: National Institutes of I tealth, 
I’llS. I IMS. 
action: Notice of proposed second 
annual update of a program to assess 
the risks of recombinant DNA research. 
summary: This notice sets forth a 
proposed second annual update of the 
program to assess the risks of 
recombinant DNA research. Interested 
parties are invited to submit comments 
concerning the plan and the desirability 
of continuing the process of annual 
updates of the plan. After consideration 
of those comments and comments by the 
Nil! Recombinant DNA Advisory 
Committee, the Director of the National 
Institutes of Health or his designee will 
publish the final update in the Federal 
Register. 
date: Comments must be received by 
February 22, 1983. 
ADDRESS: Written comments and 
recommendations should be submitted 
to the Director, Office of Recombinant 
DNA Activities, building 31, Room 4A52. 
National Institutes of Health, Bethesda, 
Md. 20205. All comments received in 
timely response to this notice will be 
considered and will be available for 
public inspection in the above office on 
weekdays between the hours of 8:30 
a.m. and 5 p.m. 
FOR FURTHER INFORMATION CONTACT: 
Additional information may be obtained 
from Dr. William J. Gartland, Jr., 
Director, Office of Recombinant DNA 
Activities, NIAID, National Institutes of 
Health, Bethesda, Md. 20205 (301^196- 
6051). 
SUPPLEMENTARY INFORMATION: 
I. Introduction 
With the issuance in December 1978 
of revised Guidelines for the conduct of 
recombinant DNA research, the 
Secretary. DHEW (now DHHS), 
requested that the National Institutes of 
Health (NIH) prepare an NIH Risk 
Assessment Plan which after review by 
the Recombinant DNA Advisory 
Committee (RAC) and publication in the 
Federal Register for comment, would be 
made final and updated annually. The 
initial proposed plan was published for 
public comment in the Federal Register 
on April 2, 1979 (44 FR 19302). Following 
analysis of public comments and those 
of the RAC, the plan was made final on 
September 13, 1979 (44 FR 53410). A 
proposed first annual update of the plan 
was published for public comment in the 
Federal Register on September 17, 1980 
(45 FR 01H74). Following the analysis of 
public comments and those of the RAC. 
the plan was made final on June 10. 1981 
(48 FR 30772). The present document is 
the proposed second annual update, 
issued for public comment. 
We stated in the first Plan issued in 
1979 and it is still our conviction that: 
The vast majority of information relevant 
to recombinant UNA risk analysis has 
already come from research not primarily 
designed to provide information on risk. This 
will undoubtedly continue to be the case. 
This information will be obtained chiefly 
from publications in the scientific literature, 
from persons witli special scientific 
knowledge, and from ongoing basic 
biomedical research. Risk assessment 
analysis will require continuing review of 
data developed in the fields of microbiology, 
infectious diseases, and related biological 
research. 
Some essential information has been, and 
will continue to be, derived from projects 
specifically designed to assess various 
aspects of potential risks associated with 
recombinant UNA experimentation. Such 
experiments will be supported by the 
Intrnmual and the Extramual programs of 
NIH. Many experiments may also be 
conducted in the private sector or may be 
funded by other ugcncies or governments. 
The essential goal of a successful risk 
assessment plan will be the development of 
means to collect, collate, coordinate, 
evaluate, and disseminate data obtained from 
all sources. 
Under the “Scientific Aspects" of the 
first Plan, it was noted that a number of 
events must occur before a laboratory 
microorganism becomes a possible risk 
to people or higher organisms outside 
the immediate laboratory environment. 
A major aspect of the risk assessment 
plan was to acquire and analyze 
information and data relevant to those 
elements for the host-vector systems 
most commonly in use. Emphasis has 
been on the prokaryotic E. coli K-12 
systems because those were, and 
remain, the systems predominately used 
by investigators. Several areas were 
identified as requiring particular 
consideration, and progress has been 
made in collecting and/or analyzing 
data on them. Before considering these it 
is worth saying that, despite intensive 
review by the RAC and NIH staff, 
several conferences and workshops to 
consider specific issues, and many 
experiments, no risks of recombinant 
DNA research have been identified that 
are not inherent in the microbiological 
and biochemical methodology used in 
such research. A synoptic report of 
progess follows. Data, reports, and other 
documents referred to are available on 
request from ORDA. 
II. Scientific Aspects 
A. Biological Activity of Polyoma — 
pBR322 Recombinants Cloned in Wild 
Type Escherichia Cali and E. Coli K-12 
One of the early concerns raised 
about recombinant DNA 
experimentation was the potential 
hazard of cloning intact animal virus 
genomes in E. coli K-12. The concern 
was that E. coli K-12 might be capable 
of producing infectious virus particles 
which then might be capable of infecting 
sites not normally exposed during 
normul infectious processes or that the 
recombinant molecule, in the absence of 
production of complete viruses, could be 
transferred to a eukaryotic cell in which 
the recombinant molecule could become 
infectious. 
Chan et al. (Science, 203. 887-892 
(1979)) and Israel et al. (Science, 203. 
883-887 (1979)) conducted experiments 
to determine whether E. coli K-12 
harboring plasmids containing a 
complete copy of the polyoma virus 
genome could cause infection in mice. 
This is a sensitive experimental system 
because mice are highly susceptible to 
infection with polyoma virus and 
develop an antibody response against 
polyoma capsid protein. In these 
experiments, E. coli K-12 containing 
polyoma-plasmid or polyoma-phage 
recombinants, cell free polyoma-phage 
particles, recombinant polyoma-phage 
DNA, or recombinant polyoma-plasmid 
DNA were fed or inoculated into mice, 
and their sera subsequently examined 
for the presence of antibody to polyoma. 
These experiments showed that none of 
the mice developed a viral infection 
after receiving E. coli K-12 carrying 
polyoma-plasmid or polyoma-phage 
DNA recombinants, or after receiving 
purified monomeric polyoma-plasmid 
DNA or purified monomeric polyoma- 
phage DNA. Infections did occur in 
control experiments when mice were 
inoculated with preparations of 
polyoma-plasimd or polyoma-phage 
DNA that had been cleaved by the same 
restriction enzymes used in their 
construction, and, therefore, contained 
unmodified polyoma DNA. 
These experiments were not able to 
assess adequately the possible transfer 
of animal virus recombinants out of E. 
coli and into susceptible mammalian 
cells because Ec. coli K-12 and its 
derivatives are not able to colonize the 
gastrointestinal (GI) tract of mice or 
humans. Consequently, Cecil Smith and 
Dr. Malcolm Martin of the Nationul 
Institute of Allergy and Infectious 
Diseases extended these studies by 
colonizing the Cl tract of conventional, 
antibiotic-compromised, or germ-free 
