Project Title:
14,
Scope:
cells,
Biological Studies of Materials Arising from Energy Operations
The Effects of Pollutants on Cells Studied in
Tissue Cultures
RX-03-06-(c)
(Cont'd.)
human alveolar macrophages, embryonic human tissue cultures,
lymphocyte cultures,
time,
These tissue cultures would be measured for:
2) growth fraction,
3)
and
1) doubling
time parameters of che cell cycle utilizing
the flow of tritiated thymidine labeled cells through mitosis, 4) the
possible induction of somatic mutations (enzymatic changes), 5) changes in
function, e.g,, effects on production of erythropoietin and other enzymes by
cultures of human renal glomeruli, and 6) cytogenic effects.
One theory on the development of emphysema suggests that a result of
pulmonary irritants is the interstitial death of neutrophils and/or macro-
phages and the concomitant release of potent destructive lysosomal enzymes
that produce microscopic foci of necrosis which, over a prolonged period,
produce emphysema (Janoff).
However, the detection of such an effect is
almost impossible by ordinary serial section of lungs because the sizes of
the lesions are too small,
It is therefore proposed that a tissue culture
system be developed to assay such effects,
The system would consist of
three Layers of different cells in soft agar:
the deepest layer consisting
of an appropriate feeder system; the next layer consisting of human pulmonary
epithelium obtained from viable fetuses; and when this culture is established,
a third layer consisting of human alveolar macrophages and neutrophils
exposed to particulate atmospheric and/or gaseous pollutants,
The basis for
this system is as follows:
if the phagocytes ingest particles for which the
q
cell is not programmed to inactivate, there is a high probability that the
result would be the death of the neutrophil or macrophage and the release
of the lysosomal enzymes (hydrolytic and oxidative) thus leading to the
death of underlying pulmonary epithelial cells,
Im principle, this could
be identified by the development of transparent plaques,
If
the
tissue culture
studies produce
positive results,
.
chronic
inhalation studies would be initiated using animals exposed to variable
concentrations of the same materials used in the tissue culture studies.
Lifetime observation of these animals would include:
1) the incidence
of pulmonary disease, 2) somatic effects, 3) pulmonary function, 4) reproductive capability, 5) the induction of dominant lethals, 6) the effects
On embryo in culture, and 7) carcinogenesis,
The proposed studies would be in collaboration with the BNL Department
of Applied Science where pollutants would be prepared for use in tissue
ggnd
in animal studies.
Dr, Michael Bender will participate for the
M@eic studies, and a scientist will be required for the study of the
embryos,
(See Continuation Sheet)
RX-391
hb 149334
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