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Federal Register / Vol. 47, No. 235 / Tuesday. December 7, 1982 / Notices 
polypeptides (including hormones) by E. 
coli K-12 should they colonize higher 
organisms. 
The workshop participants 
recommended that additional 
information should be gathered on the 
handling and absorption of polypeptides 
in normal and pathologic colon and the 
potential effects of synthetic peptides on 
the bowel itself. NLAID solicited grant 
proposals through a Request for 
Research Crant Applications (RFA) in 
November 1980. The objective is to 
determine 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. 
Six applications, received in response to 
this RFA. were reviewed by a Special 
Study Section in December 1981. 
The National Advisory Allergy and 
Infectious Diseases Council supported 
award of a project to the Medical 
College of Ohio on behalf on Dr. Murray 
Saffran. Dr. Saffran suggests a unique 
concept (bacterial degradation of azo 
bonded peptides) as a method for 
studying colonic absorption of 
hormones. Using D-argininevasopressin 
as a model peptide and employing an 
unusual strategy to deliver the active 
components to the large bowel, the 
investigators will determine if certain 
hormones can be absorbed from the 
large bowel. Vasopressin, synthesized to 
contain D-arginine, will be covalently 
coupled to a polystyrene polymer with 
an azo bond. This bond can be split only 
by intestinal bacteria and will deliver 
the vasopressin in active form to the 
large bowel after oral administration; 
the D-arginine facilitates detection. 
Further extension of the work would 
include studies on the delivery of insulin 
and growth hormone to the large 
intestine in azo-linked capsules. 
Degradation of the capsules by the fecal 
flora would release the hormones in the 
large intestine simulating release by 
recombinant DNA containing bacteria. 
A grant (Ai 18710) to support this 
project was awarded by N1AID in 
March 1982. 
F. Antibody Responses to Protein 
Produced by Recombinant Organisms 
The issue which generated the 
greatest discussion during the final 
session of the 1980 Workshop on 
Recombinant DNA Risk Assessment 
was the possible occurrence of 
autoantibodies or auto-reacting cells 
due to the production of eukaryotic 
polypeptides (including hormones) by E. 
coli K-12 should they colonize higher 
organisms. There was sentiment among 
the participants that data should be 
obtained in the interest of scientific 
thoroughness but that the scenario did 
not have a high probability of inducing 
immunologic disease. 
A Request for Proposal (RFP) was 
issued in October 1980 soliciting 
proposals from organizations having the 
capabilities and facilities to determine if 
mice can mount an antibody response to 
insulin which is being produced by 
recombinant DNA technology in E. coli 
host-vector systems. The RFP was 
published in the NTH Guide for Grants 
and Contracts and the Commerce 
Business Daily, distributed to all 
participants of the Workshop on 
Recombinant DNA Risk Assessment, 
and publicized through announcements 
to the Recombinant DNA Advisory 
Committee and the NLAID Council. 
Although seventy requests for the 
detailed RFP were received, only one 
proposal was received. This proposal 
was reviewed by the NIAID 
Microbiology and Infectious Diseases 
Advisory Committee in March 1981, and 
recommended for disapproval because 
of scientific weaknesses in the 
protocols. There are no plans at present 
to issue another RFP for this project. 
G. Cloning and Expression of DNA 
Coding for Diptheria Toxin 
The NTH has approved, on 
recommendation of the RAC, a project 
by Dr. John Murphy of Harvard Medical 
School to clone in E coli K-12 fragments 
of corynephage beta which carry the 
diptheria toxin structural gene. The first 
experiments will be risk assessment 
studies involving guinea pigs that have 
been treated with broad spectrum 
antibiotics to reduce their normal flora. 
These animals will then be fed viable E. 
coli that carry the diptheria toxin 
structural gene. These experiments may 
provide information on issues discussed 
at the 1980 Workshop on Recombinant 
DNA Risk Assessment, namely, the 
handling and absorption of polypeptides 
from the lower gastrointestinal tract and 
the effect of cytotoxins on the GI tract. 
These risk assessment experiments will 
be carried out in the NLAID high 
containment laboratory located at the 
Frederic Cancer Research Facility. 
III. Document on Evaluation of Risks 
Associated With Recombinant DNA 
Research 
In response to a proposal to convert 
the NIH Guidelines to a code of 
standard practice, a working group was 
appointed in May 1981 to review the 
status of the NIH Guidelines. During the 
summer of 1981, this group attempted to 
address the issues arising from changing 
perceptions of the hypothetical risks 
associated with recombinant DNA 
techniques. The group evaluated, 
compiled, and organized scientific 
information on risk assessment. One of 
the documents prepared by the working 
group is entitled Evaluation of the Risks 
Associated with Recombinant DNA 
Research. The document includes an 
historical overview and discusses topics 
including possible hazards, uniqueness 
of organisms created by recombinant 
DNA techniques, dissemination of 
recombinant organisms, and possible 
harm to organisms and the environment. 
The working group drew the following 
conclusions at the end of the report: 
Given the above analysis of the risks 
associated with recombinant DNA we have 
come to the following major conclusions: 
1. That accidental combinations of genes, 
rising out of "shotgun" cloning experiments 
or experiments where expression is not 
specifically engineered, are extremely 
unlikely to lead to serious problems. Both the 
barriers to expression of foreign genes in 
most organisms, the necessity for new 
activities to function as an integrated part of 
an existing pathway, and the selecUve 
disadvantage given to an organism by 
recombinant DMA inserts will interfere with 
such organisms establishing themselves in 
the environment and, thus, ultimately with 
their potential to cause harm. Therefore, for 
these experiments, the minimal controls 
associated with good laboratory practice 
should be sufficient. 
2. A particular subset of experiments may 
still pose some possibility of risk. While there 
is no evidence that this is qualitatively 
different from the risks associated with other 
kinds of genetic research, the possibility for 
improving the virulence, host range, or 
survivability of some pathogens does seem to 
exist. In most cases, in these experiments the 
problems of expression of foreign functions 
will have been bypassed, or normal functions 
will have been engineered to operate more 
efficiently. 
In many cases, even the best engineered 
strain will be at a major disadvantage in the 
environment or will require artificial 
selections to maintain recombinant DNA 
information. The issue to be faced here, 
however, is: (a) How serious it this risk? fb) 
What is the most effective, uoo-obtrusive 
mechanism for guarding against any 
untoward consequences of such work? 
Interested readers are encouraged to 
read the full report which includes an 
appendix on experiments with E. coli 
(Recombinant DNA Technical Bulletin 4, 
166-179 (1981) and Federal Register 
December 4, 1981 (46 FT 59385)). 
IV. Fundamentals for Safe 
Microbiological Research 
NIAID awarded a contract to the 
University of Minnesota to develop 
resource materials for a comprehensive 
course on microbiological principles and 
techniques for work with potentially 
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