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Protein
Synthesis
in Human
1. Effect
By
T
SEYMOUR
protein
Steroids
WERTHAMER,
HE IN VITRO
taming
medium
phoid
of
Leukocytes
and
RNA
and
CAiu.
CULTURE
has been
HICKS
of rabbit
demonstrated
synthesis.1
and
In liver
Sterols
AND
cells
in the
however,
both
RNA
suggesting
and
the
protein
target
synthesis,2
while
inhibiting
the
cell specificity
of this hormone.
of cortisol
Inasmuch
or cortisol
as cortisol
analogs,
have
been
shown
to
is well known
to be a “lympholytic
that
lymphopenia
and
cyte,
that
this
seemed
results
at a specific
cortin.5
This
level
latter
synthesis
of
human
regulator
in the
haps,
may
be
binding
its
from
inhibition
feasible.
the
inhibition
ultimately
Furthermore,
lymphocytes,
the
protein.
of the
result
have
with
that
its
normal
of
of the
therefore
level
lymphopenia.4
the possibility
protein
synthesis
ratio
the
results
MATERIALS
plasma
protein,
does inhibit
the
in
of
the
of plasma
may
act
regulation
levels
the
transprotein
of
as
per-
cortisol
investigation
to
of
the
cortisol
and
related
steroids
and
leukocytes
and purified
lympho-
the
AND
plasma
This
undertaken
cytes.
presents
increases
result
in
agent,”
lymphocytes.
of action
of
of human
mixed
paper
hormone
of DNA,3
thus
administration
its binding
if cortisol
in vitro
effect
and mechanism
sterols
on protein
synthesis
This
this
synthesis
In man,
of lymphocytic
number
of a shift
We
in a cortisol
coninhibition
of lym-
results
in the destruction
of the lymphocortisol
is found
to exist
in the plasma
and in equilibrium
point
would
suggest
control
AMARAL
LEONARD
lymphoid
to result
cells,
Lymphocytes:
first
phase
of
our
investigation.
METHODS
MateriaLi
Cortisol,
Prednisolone
and
Testosterone
respectively.
3-Methyl
Cholanthrene
Leo
Soskind
and
J.
was
purchased
chased
from
Julius
from
of
All glassware
human
blood,
used
donated
per
respectively.
International
Chemical
purchased
Cholesterol
ml.
of
Suspensions
whole
of
for preparations
by healthy
blood
),
and
Nuclear
Sigma,
Sigma
generously
specific
C14L-Ieuc1ne,
Normal
Corporation.
Human
and
Vitarine,
provided
activity
of
Eagle’s
TC
submitted
by
240
Drs.
mC/mM,
199
was
pur-
allowed
to
sediment
for
and
siliconized
Hospital,
a period
Lymphocytes
and
sterilized.
was
heparinized
of
two
hours
the upper
two thirds
of the
at 150 g. for 10 minutes.
pipette-leaving
just enough
Whole
(20
37 C.
sedimented
The
superplasma
to
at
accepted
for publication
May 15, 1969.
Director
of Laboratories,
Methodist
Hospital
of Brooklyn; Clinical
Assistant
Professor,
Department
of Pathology,
Downstate
Medical
Center,
Brooklyn,
N.Y.
CARL
HIcKs,
M.S. : Research
Assistant,
Methodist
Hospital
of Brooklyn.
LEONARD
AMARAL,
PH.D.
: Director,
Section
of Research
& Development,
Department
of
Laboratories,
Methodist
Hospital
of Brooklyn.
SEYMOUR
348
WERTHAMER,
February
Leukocytes
and incubations
was
employees
of Methodist
5 ml. pipettes.
After
the sedimentation
period,
plasma
was siphoned
off, pooled,
and centrifuged
natant
plasma
was gently
removed
with a Pasteur
in
First
from
were
Difco.
Preparation
units
Carr,
were
and
6, 1969;
M.D.:
BLOOD,
VOL.
34,
No.
3
(SEPT.)
1969
From www.bloodjournal.org by guest on June 15, 2017. For personal use only.
PROTEIN
349
SYNTHESIS
cover
the cap of leukocytes.
The
plasma
was subsequently
centrifuged
at 1500
g. for
20 minutes.
This step was sufficient
to remove
all the platelets
from the plasma.
For
the
cultures
to contain
leukocytes,
the cap of leukocytes
was genfly
suspended
in a volume
of platelet
free autologous
plasma
which
would
result
in a cell number
of 1 1 X 106 per
ml. For pure lymphocyte
cultures
the cap of cells was gently
suspended
into a final volume
of 3 ml. of platelet
free plasma,
and a pure suspension
of lymphocytes,
slightly
contaminated
by red
blood
cells was obtained
according
to the procedures
described
by Rabinowitz.#{176}
The eluted
lymphocyte
suspensions
were adjusted
to 6.0 X 106 cells per ml. of plasma.
The
use of a single
sample
of 80 ml. of whole
blood
yielded
enough
cells for any one experiment.
For those experiments
involving
the use of saline-washed
cells resuspended
in plasma
or
cells
cultured
After
placed
in
centrifuged
at
with
2 ml.
wash
procedure
0.6
was
200
ml.
cubated
For
ously
dissolved
10-6,
10-s,
are
of
or
ml.
0.6
450
angle
the
use
of
the
incubation
of
trypan
detectable
the
for
absence
both
cap
of
of
used:
cells
were
suspended
and
was
then
washed
subsequently
discarded.
This
the use of saline washed
cells
culture
tubes to a final volume
plasma,
cultures
described.
the
final
volume
subsequenfly
All experiments
and
components
and
in
saline
-2
10
M.
The
of
suspended
were
C
of
cent
or
performed
0.6
ml.
and
in-
in duplicate
earliest
response
the
use
at
culture:
0.4
0.4
25
of
the
cells
of
two
agents,
ml.
of
4
of
C.
the
experimental
final
of TC
199
or
2
per
agents
of
used
of
of
ethanol),
incubated
period
concentration
culture
cent
were
hour
(previ-
per
-2
cultures
( contain-
sterol
concentration
This
the
ml.
hormone
saline
These
and
to
2.0
ml.
a final
suspension.
37,
Period
yield
received
lymphocyte
2 hours
3 ml.
14C-leucine),
ethanol-to
controls
of
Incubation
of a final
3s
per
involving
of the
trypan
above
steps
each
at
a
incubation
at
37
was
C.
For
10-s
M.
blue exclusion
test for viability7
was routinely
carried
out after
( i.e., before
column,
after
column,
after
centrifuging
and after
). After
each of the procedural
steps,
the viability,
as judged
by the criterion
blue
exclusion,
remained
upwards
of 98 per
cent,
demonstrating
that
little
injury
had occurred.
Used
the
were
of Viability
Procedures
At
the
cells
to be
penicillin
Determination
The
the
g.,
Composition
a period
cultures
each
free
The
of leukocyte
for
represents
those
procedures
the supernatant
For the cultures
involving
plasma
was added
to the
involving
of Culture
units
and
discarded,
once.
conditions
following
cultures.
following
300
the
at 200
platelet
saline.
the
plasma,
lymphocyte
suspensions,
0.6 ml. of the suspension
2 ml. of saline
was added.
The culture
tubes
were
supernatant
cultures
with
Description
The
the
repeated
those
under
of
centrifuged
was
triplicate
ing
g.,
in plasma,
achieved
or
absence
the adjusted
tubes
and
of saline,
resuspended
of
the
obtaining
into culture
end
of
0-4
C.,
After
the
the
Incubation
incubation
of Cultures
periods,
ml.
of
0.1
N
NaOH
g.
for
10
minutes,
placed
resulting
in
the
the
in
an
supernatant
ice
bath
and
gently
suspended
in 2.0 ml. of 1 per cent acetic
acid to disrupt
the erythrocytes
centrifuged
at 200 g. for 10 minutes
and the supernatant
discarded.
Three
suesaline
washes
and centrifugations
( as before ) followed.
The pellets
of cells were
1.0
200
were
pellets
present,8
cessive
in
at
cultures
to
resuspended
centrifuged
the
cooled
dissolving
of
the
discarded,
cells.
These
the
were
then neutralized
with the equivalent
amount
of 0.1 N HCL,
placed
in an ice bath, the total
protein
precipitated
with a final concentration
of 5 per cent TCA, the precipitate
subsequently
collected
and washed
on Millipore
filters ( 0.45 u pore size ) . These
filters were then mounted
on 2 inch planchettes
and dried under
infrared
lights.
The radioactivities
of the above
were
obtained
by the use of a Tracerlab
132 M Sealer
gas flow counter
(efficiency
of 38 per
cent ) . The data presented
are the per cent
alterations
of the average
total
counts
per minute
per
set
in CPM
of duplicate
within
or
triplicate
triplicate
cultures
cultures
did
to
not
that
exceed
of
their
controls.
4 per
cent.
In
all
cases
the
variations
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350
WERTHAMER,
Fig. 1.-The
effect of steroids and sterols
on the incorporation
of 14C leucine
into
human
leukocyte
total
protein. An average
of 6.5 X 106
leukocytes
suspended
in
0.6
obtained
from
hires
of 9 different
cell
All
incubations
were
formed
molar
agent
p <
in triplicate.
concentration
in the culture
.001
for each
experiments.
formed
lOco
in
no
IP
ft
1_I
T I sicajIIllhIllUlIII
iiii-I
R E D N IS 0 L 0 N1
#{149}1R
TDIt
RUN
METHYL
,3
donors.
CH?LANTHREP4%.4
per-
The final
of each
was
series
of
I I -I
::cHoLEsTERo.L:.:::::
per-
I
I
I
I
I
40
30
20
10
0
4 C. resulted
significant
%INHIBITION
inhibitions.
CONTROL
Fig. 2.-The
effect of sterols and two concentrations
of
steroids
on the incorporation
of ‘4C leucine
into lymphocyte total protein.
Incubations
_-4
are described
in text.
Results
obtained
at 10
NI and
10
M of steroid
are presented
as
upper
and lower
bars,
respec-
ISOLO
sTERoNE:
I.’
tively.
I
:CHOLESTER9L:
I
I
30
20
per
noted
from
different
cell
bations
were
triplicate.
0
series
cent
are
obtained
-l
I
The
tions
i-4-4
IHRAt
40
I
I
i
I
cul-
Incubations
at 25 and
AMARAL
CONTROL
ml. of plasma
were
incubated
at
37
C. under
conditions
described
in the text. The per
cent
inhibitions
noted
are the
averages
HICKS,
averages
cultures
11
of
donors.
All
men-
for
each
performed
p
of
inhibi-
the
<
.001
in
experiments.
% INHIBITION
RESULTS
The
effects
cine
were
of
the
( 3-methyl
sterols
( cortisol,
hormones
cholanthrene,
into leukocyte
total protein
observed
when
incubations
are
grade.
Inasmuch
as
the
leukocyte
types,
the
inhibitions
may
above
responses
bers
of
the
agents
obtained
described
of each
the
cell
leukocyte
by
in
the
the
type
thus
population
employed.
To
methods
text.
test
)
results
)
testosterone
the
incorporation
and
of
the
14C-leu-
.
population
be
the
leading
may
this
on
inhibitory
at 37 C. ( Fig. 1 ) No inhibitions
were
performed
at 25 and 4 degrees
centriresult
to the
each
possibility,
of Rabinowitz,6
The
prednisolone,
cholesterol
were
composed
of
a summation
possibility
respond
pure
presented
is
in
of
various
of
that
distinct
a different
suspensions
mem-
manner
to
of lymphocytes,
incubated
under
in
2 indicate
Figure
cell
unknown
the
conditions
that
the
From www.bloodjournal.org by guest on June 15, 2017. For personal use only.
PROTEIN
351
SYNTHESIS
agents
inhibited
amino
acid
incorporation
creasing
the concentrations
of the steroids
cent
inhibitions
of amino
acid incorporation.
2 indicate
3-methyl
that the
cholanthrene
inhibitory
are
leukocyte
incubations.
This
of leukocyte
agents
than
population
lymphocytes.
lymphocyte,
leukocyte
The
a pure
procedures
suspension
tions
for
from
glass
of such
to
bead
eluted
and
by
and
EDTA
lymphocytes.
)
results
to incorporate
the
specific
per
the
amino
calculations
affords
into
the
type
two
non-
it has
of
alteration
for the determination
in our study,
the
cell
of these
normal
cent).
elution
acid
and
in the
the separation
of
under
the condi-
However,
that
in a gross
‘4C
suitable
employed
mathematical
( 90
neutrophil
elsewhere
columns
some
to the action
portion
of this
Rabinowitz6
would
afford
which
could
be cultured
presented
no direct
method
is presently
of neutrophils
to the agents
procedures
is the
total
protein.
Inincrease
in the per
of Figures
1 and
to testosterone
those
produced
that
resistant
largest
leukocytes
be
cells
suggests
be more
far,
the
population
both
( data
of the lymphocytes
magnitude
than
comparison
must
By
described
by
of neutrophils
employed
experience
responses
of a greater
into
lymphocyte
resulted
in no
A comparison
our
been
neutrophils
in the
protein.
ability
Although
of the response
of the following
use
deduction
of
such
re-
sponse.
A prepared
and B. The
leukocyte
total
number
determined
for
known
for
and
the
tures
Triplicate
equal
cell
preparation
known
and
of
of
response
was
A.
the
suspension
was
of leukocytes
control
number
neutrophils
determined
by
the
and
were
of triplicate
equal
cell
of
divided
equally
into
and a cell differential
experimental
prepared.
control
number.
the
The
leukocyte
cells
and
experimental
Incubations
and
leukocyte
following
cultures
mathematical
two
parts,
A
analysis
was
to
B were
lymphocyte
determination
the
agents
culof the
employed
calculations:
P
=
CPM
Y
=
The number
the per cent
of lymphocytes
of lymphocytes
in the leukocyte
culture
which
X leukocyte
number
divided
which
the
per
P X Y
Z
=
by
W
\v
=:
-
Z
X
=
Performing
ture
the
of lymphocyte
equals
the
total
total
lymphocytes
protein/lymphocyte.
counts
minute
counts
per
minute
of total
leukocyte
The
counts
per
minute
of
protein
for
each
these
calculations
their
agents
population.
total
of the
corresponding
experimental
on the non-lymphocyte
protein
contributed
( mainly
non-lymphocyte
illustrate
this
Inasmuch
teroP2
hibitions
results
protein
of the
leukocyte
of that
set.
non-lymphocyte.
and
sets allows
( neutrophils
lymphocyte
)
of
neutrophil)
population.
Table
cul-
one to deduce
portion
of the
such
experiments
and
calculations
presented
in Figure
3. A comparison
of the Figure
to Figure
2, suggests
in reference
to testosterone
and 3-methyl
cholanthrene,
the inhibitions
in the lymphocyte
population
are more
marked
than
that
observed
leukocyte
The
of total
equals
by 100.
present.
The
controls
and
effects
of the
of
used
cultures
of part
1 serves
to
are
that
noted
in the
readily
comparison.
as
cortisol,9
bind
to plasma
of amino
acid
testosterone,1#{176}
proteins,
the
incorporation
3-methyl
possibility
produced
cholanthrene1’
was investigated
by these
agents
and
choles-
that
the
is dependent
in-
From www.bloodjournal.org by guest on June 15, 2017. For personal use only.
352
WERTHAMER,
r r tu
,
D (
HICKS,
AMARAL
J
I
ISOL
IIIICORT
deduced
efsteroids
on
incorporation
of 14C leucine into neutrophil
total protein. Methods
involved
in the
fect
the
Fig. 3.-The
of sterols
and
determination
of
presented
::.cHoLEsTERoL.::::I
40
30
20
10
the
data
are
the
averages
obtained
from
the
triplicate
incubations
different
of cells
individuals.
from
3
0
% INHIBITION
Table
1.-Comparison
14C Leucine
Per
into
Cent
of the Effects
Total Protein
Inhibition
of
Cortisol
14C
of Agents
Tested
of Lymphocytes
leucine
Prednisolone
on the Incorporation
and Neutrophils
Incorporation
Produced
3-Methyl
Testosterone
of
by:
Cholanthrene
Cholesterol
Lymphocyte
21.4±1.4
14.6±1.7
17.8±1.9
17.0±2.5
22.0±4.3
Neutrophil
22
21
9
8
21
I
CONTROL
Fig. 4.-The
effect of sterand
steroids
on the incorporation
of 14C leucine
into
total
protein
by normal
human lymphocytes
cultured
in
the absence
of plasma.
Condi-
lllc
OR
ols
tions
of culture
in text.
are
from
cells
viduals.
The
are
results
I ISO
llhJIIIIIIIJIIJIIJIJJIIllhIJJJltItIUhIIllllIllIllllllHIII
L
described
presented
METHYLCHOLANTHRENE
the
averages
obtained
triplicate
cultures
of
from
3 different
mdi-
::::cHoLEsTERoL
I
I
I
I
I
40
30
20
10
0
% INHIBITION
upon
the
the
presence
inhibitory
acid
into
of plasma.
conditions,
of plasma.
The
of
three
actions
the
lymphocyte
total
protein
The
sterols,
3-methyl
still
inhibit
amino
acid
results
presented
hormones
are totally
cholanthrene
incorporation.
on
in Figure
the
4 suggest
incorporation
of
dependent
upon
and
cholesterol,
However,
that
amino
the presence
under
these
in
the
case
of
From www.bloodjournal.org by guest on June 15, 2017. For personal use only.
PROTEIN
353
SYNTHESIS
-II
CONTROL
Fig
5-The
effect
of
ster
1111111
CORTISOL
oids on the incorporation
of
14C leucine
into total protein
IsoLE
by human
lymphocytes
viously
washed
in saline
TESTOSTERONEI
cultured
ing
in
medium
autologous
preand
contain-
i
i
20
0
plasma.
40
30
0
% INHIBITION
cholesterol,
the
magnitude
in the presence
To eliminate
cells
results
in
lymphocytes
were
subsequently
cultured
pear
the
to result
of the
loss
in
that
the
of
in an increase
as
great
as
as
those
of
of the
the
in the
the
above
The
the
hormones,
experiment
results
in
Instead,
of the
of
of the
lymphocytes
magnitude
observed
washing
action
plasma.
hormones.
that
the
to the
containing
washing
actions
not
response
saline
in a medium
inhibitory
is
to Figure
2).
in these
experiments
that
the
washed
5 illustrate
prevent
inhibitions
( compare
of plasma
the possibility
in saline
in Figure
of
and
presented
saline
saline
inhibitions
does
not
washes
ap-
of amino
acid
incorporation.
Because
the
acid
amino
possibility
was
hypothesis
of the
the per cent
suiting
from
tions
in its
cortisol
Since
these
cortisol
site
are
38 per
that
of
any
two
in
their
about
and
3-methyl
equal
to
35
per
and
3-methyl
any
agents
( 10
sterols,
the
different.
the
in no
The
concen-
increase
in
significant
increase
rewhose
final
concentraM
of
lymphocyte
in
result
),
site
be
increase
to
2
of
and
may
the
tested
)
over
action.
cholanthrene
as only 3-methyl
results
in an
cent
since
Figure
of
inhibition
hormones
observed
previously
cholanthrene
action
that
( see
use
the
the
of inhibition
been
a difference
site of action
or testosterone,
3-methyl
of
is
has
6 suggest
increases
and
tory
indicates
( about
expected
site
experiment
those
of
between
their
which
were
Figure
in
this
culture
others
either
the
in
mediation
different
that
hormones
alone
sented
which
is
inhibition
produced
the simultaneous
in that
each
by
investigated
tested
tration
l)y
manner
incorporation
that
The
differs
produced
results
from
pre-
all
the
cholanthrene,
in conjunction
with
increase
in the per cent inhibition.
the
sum
of the
for
3-methyl
cent
testosterone
cholanthrene
per
cent
),
respectively
must
be
inhibitions
cholanthrene
different
the
from
and
inhibithat
of
hormones.
DI5uSSI0N
The
results
analog
rene,
and
its
lymphocyte
presented
prednisolone
cholesterol,
and
leukocyte,
in
and
inhibit
this
paper
suggest
testosterone,
and
the incorporation
total
protein.
That
that
the
the
sterols,
of amino
these
are
hormones,
cortisol,
3-methyl
cholanthacid
into
human
indeed
inhibitions
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354
HICKS,
WERTHAMER,
I
CONTROL
lLc9.R
AMARAL
OLIIJIJIIJIIIIIII
TIS
IT E STOSTERON
E
3
_________________
:#{149}
c HO L E S I
F RO L
:
::::I
Fig. 6.-The
combinations
ol
____________________
effect
on
corporation
total protein
IlllIIlHhlIlIIIllIIHIllIHIIllIllllIlllllhIII
___________________
of leucine
by human
phocytes.
The
centration
culture
of
was
description
(iiIIIiiIILLLt%III1JIHIIIIIiII!iIiiJI1
is
of sterthe in-
final molar
any
10.
the
of
con-
agent
per
Detailed
experinent
in the
provided
into
lyni-
text.
.
_
I
I
I
40
30
20
I
I
10
0
% INHIBITION
and
not
the
caused
by
from
1
the
)
the
loss
blue
incubation
2
)
agents
or
and
that
precipitation
of
1 per cent
acetic
loss of labelled
studies
protein
have
cell
the
cell
injury,
manner
death
or
of handling,
counts
of
agents
hemagglutinin.
ment
with
the
lysis,
is evident
cultures
employed
serum
total
steroid
dependence.
precursor,
for
the
cell
washes
following
were
neither
of
that
lymphoid
inhibits
the
the
inhibitory
effect
Interestingly,
cholesterol,
cortisol1
cells.
Furthermore,
responses
of
possibility
that
produced
in
inhibit
all
sterols
normally
in-
our
the
and
in-
indicating
hydrocortison&
prednisolone14
the
lymphocyte
as
to
phyto-
are, in this respect,
in agreeet al.’ found
no dependence
by
cortisol
studies,
synthesis
whose
discarded
yielded
no activities
these
procedures.
The results
obtained
in our study
cited.
However,
whereas
Makman
those
upon
all
acid supernatant
protein
during
demonstrated
synthesis
hydrocortisone,15
gesting
the
cytolytic.
Previous
as
from
or
tests
indicate
TCA-tungstate
inhibit
resulting
employed
exclusion
periods
eluding
the
no detectable
well
radioactivity
the
following:
Trypan
jurious
of
either
we
have
both
testosterone
of lymphocyte
nuclear
structure
observed
and
protein
is not
too
a
the
sugdis-
may
have
the same
inhibitory
effects.
This
suggestion
appears
plausible in that 3-methyl
cholanthrene
also has been
noted
in our studies
to have
such
an effect
( although
it apparently
acts at a different
site).
Transcortin,
a cortisol
binding
protein
exists
at a molar
concentration
of
similar
about
and
10-i
Doe
M
)
.
in normal
Their
studies
plasma
have
( calculated
shown
that
from
this
the
protein
data
binds
presented
the
by
hormone
Seal
at
From www.bloodjournal.org by guest on June 15, 2017. For personal use only.
PROTEIN
355
SYNTHESIS
equimolar
concentrations.
From
the
binding
constant
other
studies,’6
the final concentration
of cortisol
binding
is of the order
of 10 M. At a concentration
enhance
of plasma,
the
inhibition
value
did not produce
suggest,
in the
light
incorporation
by
A similar
binding
site
is not
when
in
Our
steroids
with
These
suggest
results
means
In
therefore
of a different
conclusion,
The molar
of cortisol,17
synthesis.
than
that
increased
occur
in
of such
result
of the
affinities
to difcompete
for
if the
would
steroid
twO
of
the
above
3-methyl
of
complexes
in
the
protein
synthesis
by
present
protein
is normally
would
result
of lymphopenia
protein
in-
incorporation
lymphocyte.
condition
of lymphocyte
an
observed
in
of plasma.
of the
binding
protein
of cortisol
the
by the
when
resulted
inhibits
lymphocyte
since
produced
cholanthrene
was also
the absence
site
that
effect,
a difference
action
that such a complex,
normally
as a regulator
of lymphocyte
and
inhibition
that
cholanthrene
suggests
may
act
of
steroids,
at a different
inhibition
suggest
site
never
exceeded
results,
3-methyl
and
complex
system,
testosterone.
different
testosterone
a common
amount
of the cortisol
therefore,
administration
formation
as the
that
vitro
protein
vascular
acid
complex.
However,
present,
of
not
of amino
and
with
and
3-methyl
cholanthrene
acid
incorporation
in
mechanism
the
cortisol-plasma
in the human
are
suggest
either
inhibition.
amino
inhibition
prednisolone
and
in the absence
our results
a transcortin-cortisol
known.
these
cultures
Unlike
these
conjunction
the
these
at different
sites,
it would
be expected
results
in no increase
in its inhibitory
results
crease
in the present
our
studies
to inhibit
by
for
presently
both
of action.
per cent
inhibition
in
more
potent
inhibitor.
present
that
via
apply
were
to inhibit
in either
steroid
noted
increase
also
data,16
mediated
demonstrated
to bind
Whether
or not cortisol
sites
protein
complexes
since
an increase
in their
is
by
for the saturation
M, cortisol
did
at 10 M. Since
cortisol,
of amino
acid incorporation,
transcortin
may
has been
proteins.10
transcortin
any
of the
lymphocytes
mechanism
Testosterone
ferent
plasma
observed
inhibition
provided
required
of 10
higher
in the
would
synthesis.
SUMMARY
The
iii
vitro
effects
steroids,
cortisol,
rate popultions
It
has
been
synthesis
ably
shown
under
mechanism
as
of inhibition
indicating
that
these
of the
a protein
cholesterol
amino
acid
of
sterols,
all
and 3-methyl
incorporation.
factor
cholanthrene,
However,
is lower
than
a partial
plasma
port
the hypothesis
steroid
hormones
The
physiologic
to the condition
that
occurs
used
It
hormones
binding
role of the
of lymphopenia
result
has
also
requires
in
order
cholanthrene
in
been
the
to
the
inhibition
shown
that
presence
achieve
and
synthesis
in sepabeen
investigated.
of
the
of plasma,
its
effect.
protein
inhibitory
presumThe
sterol,
in the absence
of plasma,
still inhibit
in the case of cholesterol,
the magnitude
that
observed
dependence.
the
only
3-methyl
testosterone
on protein
and leukocytes
has
agents
conditions.
steroid
and
cholesterol
prednisolone
and
of human
lymphocytes
inhibition
when
the
in the presence
of
The results
presented
of lymphocyte
steroid
is bound
plasma
protein-cortisol
in man is discussed.
plasma,
perhaps
therefore
sup-
protein
synthesis
by
to a plasma
protein.
complex
and
its
relation
a
From www.bloodjournal.org by guest on June 15, 2017. For personal use only.
WERTHAMER,
356
SUMMARIO
Esseva
investigate
e de
le
effectos
steroides-cortisol,
separate
agentes
in
IN
vitro
de
prednisolona,
populationes
usate resultava
e 3-methyl-cholanthrena-
testosterona-super
le
hormones
rob
steroide
hypothese
que
occurre
physiologic
del
solo
complexo
de lymphopenia
in humanos
We would
Drs. Leonard
like to thank
Ornstein
and
e leucocytos
del synthese
le
synthese
de
proteina
in
monstrate
que omne
le
iste conditiones.
Esseva
etiam
monstrate
que
le mechanismo
inhibitori
del hormones
steroide
require
le presentia
de
plasma,
presumitemente
como
un factor
de ligation
proteinic
pro effectuar
iste resultato.
Le sterol-cholesterol
e 3-methyl-cholanthrena-in
le absentia
de plasma
inhibiva
ancora
le
incorporation
de amino-acido.
Tamen,
in le caso de cholesterol,
le magnitude
del inhibition
esseva
plus basse
que illo observate
in le presentia
de plasma,
possibilemente
in consequentia
de un partial
dependentia
ab le plasma.
Le
resultatos
hic
presentate
supporta
consequentemente
de lymphocytos
in le inhibition
AMARAL
INTERLINGUA
steroles-cholesterol
e
HICKS,
le
inhibition
quando
de
human.
Esseva
de proteina
sub
del
synthese
le steroide
proteina
lymphocytic
es ligate
a tin
e cortisol
e su
plasmatic
de
proteina
proteina
de
relation
per
plasma.
con
Le
le condition
es commentate.
ACKNOWLEDGMENTS
Miss Joanne
Gallagher
for her excellent
Stanley
L. Lee for their valuable
advice.
technical
assistance,
and
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From www.bloodjournal.org by guest on June 15, 2017. For personal use only.
1969 34: 348-356
Protein Synthesis in Human Leukocytes and Lymphocytes: 1. Effect of
Steroids and Sterols
SEYMOUR WERTHAMER, CARL HICKS and LEONARD AMARAL
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