From www.bloodjournal.org by guest on June 14, 2017. For personal use only.
A Comparison
of
Properties
Factors
By
E
of
cysts
and
ation
red
have
an
been
elevated
in
renal
man
mass
and
usually
oxygen
or cyst
erythrocytosis.
cases,
of
certain
extracts
Several
substances
are
include
known
to
produce
erythropoietin,12
mones,15
decade,
corticosteroids,1#{176}
dinitrophenol,14
considerable
evidence
indicates
the physiologic
been
purified,
with
protein
acid2#{176}
and
The
nature
has
cysts
stimulation
it has been
a molecular
the
protein
that
cerebellar
hemangioblastoma
of
an
lating
bellar
antibody
the
cyst
mobility
to erythropoietin
on
four
biological
materials
shown
are
containing
of
by
by
if
not
sialic
the
with
cyst
from
fluid,
reference
cyst
with
treatment
and
in-
fluid
and
in
the
size
urine
erythropoiesis-stimu-
renal
compared
tumors
radiation
identical
derived
erythropoietic
AND
requires
renal
similar,
serum,
were
effect
from
erythropoietir
investigation,
their
which
produced
anoxic
the
and
hor-
In the past
major
factor
this substance
has
that it is a muco-
factors
serum,
fluid
alcohol.17
is the
previously
present
MATERIALS
The
fluid
experimental
activity.
have
to
in anemic
hemangioblastoma
of
the
shown
androgenic
Although
means
factors
cyst
In
present
electrophoretic
We
25,000-35,000)
patient.’9
factors
been
the
in
25,OOO4O,OOO,I8.19
erythropoiesis-stimulating
weight
anemic
of
for biological
characterized.
activation
(molecular
weight
indirect
erythropoiesis-stimulating
been
blood
have
hormone13’14
and
batyl
that erythropoietin
of erythropoiesis.12
determined
by
structure2’
of the
not
cyst
erythrocytosis
thyroid
These
not
associ-
In the majority
a remission
of
in
or
this
white
24a7
animal.
in
tumor
with
a normal
saturation.
results
the
erythropoiesis-stimulating
have
various
cerebellar
Patients
11
cell
with
association
hypernephromas,38
and arterial
of the tumor
In
ir
cysts,”2
cell count,
platelet
count,
these
patients,
resection
to contain
Chemical
A. Ww.NN
AND THOMAS
noted
including
hemangioblastomas,91#{176}
and
Erythropoiesis-Stimulating
from
Different
Sources
has
and
Physical
F. ROSSE
WENDELL
RYTHROCYTOSIS
tumors
Some
trypsin,
and
to
ceretheir
sialidase,
and
activity.
METHODS
erythropoiesis
stimulating
activity
used
in
these
studies
were:
(a) Anemic
serum-Blood
was removed
under
sterile
conditions
from a patient
with
chronic
lymphocytic
leukemia
and red cell aplasia
at a time when
her hemoglobin
was 6.7 Cm. per cent.
The blood
was centrifuged
and the serum
removed
and
frozen.
The whole
serum
from this patient
was found
to contain
9.7 units
of erythropoietin/
ml. as defined
by Coldwasser
and White.22
(b) Anoxic
serum-Serum
was likewise
obtained
from
a patient
with
transposition
of the great
vessels.
The patient
had marked
arterial
oxygen
desaturation
(68 per cent)
and polycythemia
(hemoglobin
22.3 per cent).
His whole serum was found to contain
0.52 units of erythropoiesis-stimulating
activity
per
From Department
lism Service,
National
Submitted
Jan.
27,
of
Health,
Education,
and Welfare,
Cancer
lnstitute,
National
Institutes
1964; accepted
for publication
Mar.
Public
Health
Service,
Metaboof Health,
Bethesda
14, Md.
7, 1964.
739
BLOOD,
VOL.
24,
No.
6 (Drcm.ii),
1964
From www.bloodjournal.org by guest on June 14, 2017. For personal use only.
ROSSE
740
ml.
Renal
(c)
cyst
mild
fluid-Fluid
tient
with
units
of erythropoiesis-stimulating
fluid-Fluid
aspirated
normal
cyst
fluid
The
obtained
(Hct.
54).
The
activity
from
from
a single
fluid
was
yellow,
(d)
Cerebellar
per
a cerebellar
cyst
ml.
in a patient
unilocular
who
WALDMANN
renal
clear,
cyst
and
of
a pa-
contained
0.98
hemangioblastoma
had
polycythemia
cyst
(Hct.
63),
arterial
oxygen
saturation
( 95 per cent ) . #{176}Using the present assay system, this
found
to contain
14.9 units of erythropoiesis-stimulating
activity
per ml.
erythropoiesis-stimulating
activity
of samples
was determined
in the polycythemic
and
was
assay
1Th)USC
was
polycythemia.
AND
previously
injected
intraperitoneally
pension
of
described.23
on
homologous
Female
days
red
1 and
blood
NIH
2 of
cells.
On
strain
the
mice
assay
day
5,
with
an
weighing
0.7
additional
18-22
ml.
of
an
0.5
ml.
Cm.
80
of
per
this
were
cent
sus-
suspension
was given.
On days 5 and 6, 0.5 ml. of test material
was injected
subcutaneously
into each
mice. Saline was injected
into 8 mice as a negative
control.
On day 7, 0.5 c. Fe59
citrate
in 0.15 NI NaCI was injected
intraperitoneally.
On day 9, the mice were
bled by
decapitation
into tared
bottles
which
were
then reweighed.
The radioactivity
due to Fe59
was determined
in a well-type
gamma
ray spectrometer.
The
per cent
incorporation
of the
isotope
in 1 ml. of peripheral
blood
was determined
by comparison
with an appropriate
of 6-8
standard.
A
dose
response
curve
performed
with
standard
material
obtained
from
the
Hematology
Study
Section,
National
Institutes
of Health,
showed
a linear
relationship
between
dose
of erythropoietin
and the erythropoietic
response
between
.24 and 2.4 units
of the hormone.
The minimum
detectable
dose tested
was 0.03 units
which
caused
an
iron-59
incorporation
ml. in
diluted
animals
with
of
0.41
receiving
0.15
per
cent
saline.
NI NaCI
and
compared
Samples
to
an
incorporation
containing
more
of
than
2.4
0.09
units
per
per
cent
ml.
per
were
reassayed.
Electrophoresis
Zonal
electrophoresis
modified
by Fahey
( polyvinyl
chloride,
in
supporting
was
matrix.
each
narrow
groove
ma.
was
eluted
with
concentration
at 280
Undiluted
cut
applied
segments
sity
of
cut
which
is 5.5
in the
for
18
hours.
to
the
method
of
Mi.iller-Eberhart24
as
of
Geon
( polyvinyl
copolymer
particles
wide.
12 cm.
The
Two
from
to
the
Geon-Pevikon
four
ml.
of
the
sample
were
applied
cathodal
end
of the
unit.
A current
blocks
were
then
divided
into
in
a
of 50
1-.4
cm.
to the direction
of the electromotive
force.
The protein
was
using 5 ml. per cm. segment
of the block.
The relative
protein
from each segment
was found
by determining
the optical
den-
in a Beckman
aliquots
cm.
matrix
perpendicular
0.15 M NaC1,
of the eluate
mz
according
chloride
particles)
and Pevikon
) in equal amounts
were
used
Four
hundred
and
eighty
Gm.
of the mixture
was equilibrated
with
barbital
buffer,
pH 8.6, ionic strength
of 0.075,
and was placed
in a
chamber
36.5 cm. long which
was divided
longitudinally
into 4
300 ml. of sodium
Lucite
electrophoresis
subunits,
performed
and McLaughlin.25
polyvinyl
acetate
I)U
these
spectrometer.
eluates
were
tested
for
erythropoiesis
stimulating
activity
in
the assay
described
above.
The electrophoretic
mobility
of the erythropoiesis-stimulating
activity
was compared
with the mobility
of proteins
whose
electrophoretic
characteristics
were
known.
To do this, albumin,
haptoglobin
(an a2-$ globulin)
and transferrin
(a
globulin)
were
labeled
prior
to electrophoresis
with
Bromphenol
Blue,
hemoglobin
and
Fe59 ciLrate
(0.5 sc.) respectively.
The position
of albumin
was identified
after
electrophoresis
by the position
of the blue band
near the anodal
end as determined
either
by inspection
or by determining
the peak optical
density
of the eluates
read at 600 m
(optimum
for bromphenol
blue).
The position
of the maximum
optical
density
due to bromphenol
blue corresponds
to a peak of protein
concentration
(fig. 1, top panel).
The position
of the haptoglobin-hemoglobin
complex
following
electrophoresis
was similarly
identified
by
the red band(s)
as noted
on inspection
or determined
by the peak optical
density
at 412
The
clinical
course
of this patient
f Obtained
from Goodrich
Chemical
tObtained
from
Stockholm’s
has been
Company,
Superfosfat
previously
Akron,
Farbriks
A-B.,
reported
Ohio.
Stockholm,
in full.10
Sweden.
From www.bloodjournal.org by guest on June 14, 2017. For personal use only.
PROPERTIES
OF ERYTHROPOIESIS-STIMULATTNG
RI g
NEJ
741
FACTORS
ii
II’’
U
Li
U)
E
E
E
O1J02
o-x
ddo
a
0
I’.
oc
.<
-icr,
U)
-<
zw
o
Id
LI
I.->.
<0
o-J
0
1)
z
ORIGIN
‘i-CATHODC
CCNTIMCTERS
FROM
ORIGIN
ANODE-
comparison
of the electrophoretic
mobility
of erythropoiesis-stimulatin anemic
serum,
hypoxic
serum,
renal cyst
fluid, and cerebellar
hemangioblastoma
cyst fluid. Migration
from the point of application
(origin)
is shown
on the abscissa.
The relative
protein
concentration
of segments
taken
at centimeter
intervals
from the origin
are shown
for a typical
run by the solid
line in the top
panel.
The position
of the ervthropoiesis
stimulating
activity
iS shown
in black,
of
albumin
in cross-hatched
areas
(determined
by spectrophotometr
in the top panel
and by inspection
in the lower
panels),
of hemoglobin-heptoglobin
complex
in the
dotted
areas
(spectrophotometrically
in the top panel,
by inspection
in the lower
panels),
and
of transferrin
labeled
with
Fe59
citrate
by the
broken
line in the top
panel.
Fig.
1.-A
ing activity
(optimum
for hemoglobin).
The position
of transferrin
was determined
in one case by
determining
the peak
of radioactivity
in the eluate
due to Fe59 as monitored
in a welltype single
channel
y-ray
spectrometer.
In order
to ascertain
that the relative
mobilities
of proteins
on Ceon-Pevikon
were
the
same as those
commonly
seen in other
technics
of zonal
electrophoresis,
agar gel electrom
From www.bloodjournal.org by guest on June 14, 2017. For personal use only.
ROSSE
742
+15
Fig.
Pevikon
+11
WALDMANN
#{149}-1 -5
+4
+8
AND
ws
2.-The
block
electrophoretic
mobility
on agar
gel of proteins
eluted
from
a Ceonafter
electrophoresis
conipared
to the electrophoresis
pattern
of whole
serum
( WS ) . The distance
in centimeters
from
the
origin
at which
the
segment
was taken is shown b’ the numbers;
positive
nunibers
indicate
that the segment
was
in the anodal
direction
from the origin,
negative
numbers,
in the cathodal
direction.
In the agar gel electrophoresis
shown
here,
the anode
was
at the top of the plate.
phoresis
plates
was
in
verona!
18
hours.
the
buffer
cathodal
at
upon
and
figure
2,
of
in
eluates
to
8.6;
the
melted
plate.
relative
segments
was
filter
were
a current
the
1 per
mobility
agar.
to
gel
cut
in
samples
paper
fitted
of
ma.
The
36
plates
the
with
allowed
x 1 mm.
on
of
thickness
Representative
of
with
agar
or
and
thickness
stained
the
the
froni
uniform
7 mm.
agar
chamber
the
dried
the
pH
using
covered
holes
end
either
a humidified
ing
were
Rectangular
applied
in
performed
3 x 5 inches
and
the
the
of
at
for
with
cent
amido
black
in 2 per cent
of the proteins
on Geon-Pevikon
equilibrate
3.5
for
cm.
from
electrophoresis
The
applied
treated
to
a point
block
holes.
block.
Glass
par cent agar
11/2
plates
agar
from
was
the
the
into
were
Geon-Pevikon
a solution
plates
were
were
placed
1 #{189}-3 hours,
2
per
acetic
cent
acid.
agar
and
dependacetic
As
gel
acid,
shown
appear
in
to be
same.
Enzymic
Reactions
Sialidasea
with
tivated
method
each
of
of clostridial
the materials
origin,
in
free
order
the erythropoiesis-stiniulating
described
by Blunthcrg
#{176}Kindlysupplied
Diseases,
National
by Dr.
Institutes
of
to
non-specific
determine
activity.
and
Leonard
of
Health
The
Warren,26
Warren,
and
proteolytic
activity,
was
the removal
of siahic
whether
reaction
using
National
from
1-6
was
carried
ml.
of test
Institute
Behringwerke,
for
on
according
material,
Arthritis
Marburg-Lahn,
incubated
acid inacto
depending
and
Metabolic
Germany.
the
From www.bloodjournal.org by guest on June 14, 2017. For personal use only.
PROPERTIES
OF
ERYTHROPOIESIS-STIMULATING
upon
the erythropoiesis-stimulating
ther sialic acid was produced
method
of Warren.27
Controls
743
FACTORS
activity
titer. The reaction
was
hours
at 37 C. ) was measured
each
incubation
were
treated
( 6-10
for
continued
by the
identically
until no furthiobarbiturate
except
that
saline instead
of sialidase
was added
to the mixture.
Both control
and test mixtures
were
assayed
for erythropoiesis
stimulating
activity
in polycythe1liic
mice.
Tryptic
digestion
of each of the materials
was carried
out using twice
crystallized
trypsin
of bovine
pancreatic
origin.0
The course
of the reaction
was
monitored
by precipitating
the protein
of aliquots
of the reaction
mixture
with 5 per cent
perchloric
acid
and
noting
the optical
density
of the supernatant
at 280 nij. in the Beckman
DU spectrometer.
The
reactions
were continued
until there
was no further
increase
in the optical
density
at this
wave
length.
Controls
for each
incubation
were
treated
identically
except
that saline
instead
of trypsin
was added
to the mixture.
Both control
and
test
mixtures
were
assayed
for erythropoiesis
stimulating
activity
in the polycythemic
mouse
assay as described
above.
Immunokgic
Reactions
An antibody
to human
urinary
erythropoietin
was prepared
l)y a modification
of the
method
described
by Schooley
and Garcia.28
An extract
of urine
from the aplastic
patient
described
above
was made
according
to the method
of Gordon.29
The lyophihized
powder
was reconstituted
to a concentration
of 1.5 mg./ml.
At the onset
of immunization,
2 ml.
of this mixture
was injected
with 2 ml. of Freund’s
complete
adjuvant.
Two weeks
later
a further
2 ml. was injected
with 2 ml. of Freund’s
incomplete
adjuvant.
Two weeks
later,
a progressive
intravenous
immunization
with increasing
amounts
of alum-precipitated
urine
extract
was begun
according
to the plan of Kabat
and Meyar.#{176} Five days after the completion
of a three-week
course,
serum
was harvested.
Two of six rabbits
so treated,
developed
a factor
in the serum
which
destroyed
erythropoietin
activity
in serum
of anemic
patients.
This factor
was found
to migrate
as a gamma
globulin
on zonal
ehectrophoresis
performed
as described
above,
was destroyed
by heating
to 65 C. for two
hours,
but
was
stable
for one year when kept frozen.
The serum
was thus considered
to contain
antibody
to
erythropoietin.
The ability
test material
rum and the
overnight
at
performed
from
of this antiserum
to neutralize
the erythropaiesis
stimulating
activity
of the
in vitro was determined
by mixing
equal
portions
of the immune
rabbit
sematerial
to be tested,
incubating
the mixture
for
1/2 hour
at 37 C. and
then
0 C. Any precipitate
formed
was removed
by centrifugation.
Controls
were
in
which
rabbits
the
antiserum
immunized
with
erythropoiesis-stimulating
in
an
activity
reaction
mixture
extract
the
of urine
that
of
reacted
the
was
did
replaced
not
mixture
by
contain
was
saline
or
serum
erythropoietin.
assayed
as
The
described
above.
RESULTS
The
electrophoretic
stimulating
mobility
factors
cerebellar
from
active
regior.
In
case,
subtracted
the assay
from
animals
the
renal
fluid
cyst
that
above
The
antiserum
each
and
of saline
effect
total.
injected
the
case
#{176}Obtained from
the
anoxic
controls
biological
procedure
Worthington
anoxic
of
serum,
is shown
in figure
Although
the incorporation
with
the
eluates
of the
of reaction
on
fluid
block
the
erythropoiesis
serum
(p
with
is low,
fluid,
renal
cyst
1. The
mobility
in each
of the tested
materials
is in the
Fe59 incorporation
of the saline
controls
the
the
Geon-Pevikon
serum,
cyst
factor
each
In
anemic
hemangioblastoma
biologically
of
and
of the
a2-globulin
has been
of Fe5#{176}
into the blood
of
a2-globulin
sections
of the
in each
case,
it was
significantly
<0.01).
trypsin,
activity
completely
Biochemical
sialidase
is shown
and
rabbit
in table
destroyed
Corporation,
anti-erythropoietin’
1, 2 and
the
Freehold,
biological
New
3 respectively.
activity
Jersey.
of
From www.bloodjournal.org by guest on June 14, 2017. For personal use only.
744
ROSSE
Table
of Erythropoiesis-Stimulating
Reaction
with Sialidase
1.-Inhibition
Assay
Source
Activity
Saline
-
control
#{176}Standard error
Table
Results
% incorporation
of blood
(transfused
of the
2.-Inhibition
mean
shown
Fe
enzyme
4.55 (0.34)#{176}
1.00 (0.27)
0.16
0.08
(0.02)
(0.01)
5.39
1.03
(1.11)
0.19
(0.02)
(0.56)
0.07
(0.01)
0.15
(0.01)
in parentheses.
Activity
Source
of ErythropoiesisStimulating
Activity
Without
Anemic
serum
Anoxic serum
Cerebellar
hemangioblastoma
cyst fluid
Renal cyst fluid
control
of the
mean
shown
the tested
materials.
Reaction
extract
of normal
urine
did not
by
Trypsin
Results
I % incorporation
Fe”
of blood
(transfused
mouse)]
Assay
Saline
in 1 ml.
mouse)]
enzyme
of Erythropoiesis-Stimulating
#{176}Standard error
by
With
Without
Anemic
serum
Anoxic serum
Cerebellar
hemangioblastoma
cyst fluid
Renal cyst fluid
WALDMANN
Activity
of Erythropoiesis.
Stimulating
AND
enzyme
Digestion
in 1 ml.
With
enzyme
5.62
(0.55)#{176}
0.11
(0.01)
2.05
(0.66)
0.08
(0.01)
7.46
( 1.15)
1.87
(0.25)
0.06
0.07
(0.01)
(0.01)
0.05
(0.01)
_________
in parentheses.
of the materials
with
a rabbit
affect
the biological
activity.
antiserum
to
an
DIscUssIoN
Although
of great
the
bits31’33’34
mates
hormone,
biological
and
of
the
erythropoietin,
activity
have
sheep’9’32
and
molecular
the
preparation
a high
phoretic
in the
of
reflect
this
from
sort
the
the
the
region
present
poiesis
stimulating
a2-globulin
region
rectly.
Since
ations
in
the
whole
by
studies,
factor
in each
by determining
serum
of the
the
or cyst
electrophoretic
fluid
mobility
have
biologically
erythropoietin
groups
the
and
by
is made
the protein
that
of
content.31’32
has been
in the
molecule.
used,
the
region
of
the
by
erythro-
to be in the
activity
di-
possibility
procedures
with
The
electrofound
to be
a2-globulin
tested
was found
of the biological
be
esti-
made
active
materials
mobility
purification
Some
is a mucoprotein
mobility
could
rab-
been
electrophoretic
by
of anemic
patients.29
The assumption
the majority
of
and hexoseamine
purified
preparations
some
concentrates
serum
of anemic
of the
that
purified,
the
erythropoietin
characteristics
indicated
beeti
from
purified
preparations.
characteristics
of
have
ai-globulin32’34
In
urine
of
sialic
acid,
carbohydrate,
mobility
of these
partially
others.31
not
prepared
characteristics
analysis
of these
partially
the chemical
and physical
Analyses
has
been
of
was
alterelimin-
ated.
Information
about
the
nature
of
erythropoietin
molecule
has
also
been
ob-
From www.bloodjournal.org by guest on June 14, 2017. For personal use only.
PROPERTIES
OF ERYTHROPOIESIS-STIMULATINC
Table
3.-Inhibition
of
Erythropoiesis-Stimulating
Anti-Erythropoietin
0.13
(0.01)
(0.42)
0.04
(0.01)
5.67
2.66
(0.81
(0.57)
0.07
(0.01)
0.03
(0.01)
Saline
0.09
(0.01)
upon
the
control
of the
noting
weight
factors
approach
erythropoietin
the
anoxic
human
from
anemic
serum,
biological
of
on the
rats
neutralizes
mice,
rabbits,
the
antiserum
to human
of
human
found
in
the
that
and
not
this
serum
From
these
poiesis
serum
from
found
in
in renal
an
the
polycythemia
gioblastoma
seen
is
of
in
studies
human
anemic
demonserum,
have
the
same
anemic
association
probably
due
as that
that
cerebellar
to
fluid
and
erythropoietic
identical
which
the
and
frogs.
erythro-
cyst
to
the
the
sources
stimulating
hemangioblastoma
cysts
of
ac-
that
indicates
of birds
substance
and
anemic
determinants
factor
with
renal
the
secretion
renal
and
pathologic
antigenic
Thus,
anoxic
studies,
erythropoietic
from
rabbit
the
This
destroy
“normal”
patient.
Garcia
erythropoietri
the
cyst
quail.36
if not
may
Thus,
of serum
from
in the present
of anoxic
the
antiserum
urinary
neutralized
common
is similar,
an
human
stimulating
fluid,
specific
and
both
from
share
patient
serum
present
fluid
of
structure2’
of proteins.
activity
Likewise,
does
not
Japanese
it is clear
cyst
anoxic
to
as well
erythropoiesis
considerations,
found
this
activity
protein
cyst
a single
serum
antiserum
of anoxic
mammals
in the
present
from
similarity
anoxic
cysts
other
by
erythropoietin
and
factors
from
the
cerebellar
antiserum
urinary
erythropoiesis-stimulating
man
and
the
Likewise,
biological
upon
the erythropoietic
sheep
and man.
anemic
However,
present
fluid
immunologic
hemangioblastoma
serum.
fluid.19
the
molecule.2#{176} The
activity
the
have
cyst
)
that
is equal
to a molecular
for erythropoiesis-stimu-
that
factor(s
biologi-
demonstrated
cerebellar
show
is dependent
biological
to demonstrate
in
treatments
previously
activity.
rabbits
or anemic
are
to
cyst
Schooley35
in
and
others
renal
made
tivity
fluid
by
acid
and
rat
cyst
sources
of sialic
destruction
cerebellar
have
portions
of the molecule
and
that this size is the same
renal
used
for
activity
or chemical
we
erythropoiesis-stimulating
requirements
used
of physical
active
)
in parentheses.
approach,
been
presence
that
shown
effect
this
from
has
strate
mean
the
Using
biologically
of 25,000-35,000
The
antibody
1.61 (0.60)#{176}
of
be
With
2.83
by
and
antibody
Anoxic serum
Cerebellar
hemangioblastoma
cyst fluid
Renal cyst fluid
activity.
lating
-
Fe” in 1 ml.
mouse)
I
Anemicserum
#{176}Standard error
size
Without
with
Antibody
Results
I % incorporation
of blood
(transfused
Assay
tamed
Activity
Neutralizing
Source
of ErythropoiesisStimulating
Activity
cal
745
FACTORS
fluid,
and
erythropoietin
pathogenesis
of
and
cerebellar
erythropoietin
by
the
hemanthese
lesions.
It is not
has
been
surprising
shown
that
in
erythropoietin
experimental
animals
is found
that
in renal
the
kidney
cyst
fluid
is a major
since
source
it
From www.bloodjournal.org by guest on June 14, 2017. For personal use only.
746
ROSSE
of erythropoietin
produced
able
evidence
indicated
in response
to anemia
the
hormone
with
fluid
cyst
are
of the von Hippel-Lindau
seek the extra-renal
sites
might
and
be the
ovarian
stance,5
adrenal
tumors’2
mors
been
secrete
poietin
and
are
starved
the
poiesis
poietin
elaborated
produced
certain
other
the
factors
present
cerebellar
was
study
.
we
by
are
erythremia
secondary
thropoietin
in rabbits
activity
was
to an extract
materials
share
stimulating
fluid,
the
cysts and
erythropoietin
and
renal
with
sialidase
with
similar
In
le
presente
patientes
de
con
studio
stimulation
cyst
of the
(a)
anemia
nos
not
erythrohormones,
erythro-
the same,
mechanism
as erythroby which
study.
of each
on
zonal
trypsin;
fluid
appear
ha
aplastic,
certe
le
(b)
and
that
these
The
erymade
these
the
erythropoiesiscerebellar
hemangio-
similar,
if not
in association
to be
ma-
due
identical.
with
to the
renal
secretion
of
INTERLINGUA
comparate
erythropoietic
four
by an antiserum
This
implies
that
seen
appears
implies
mobility.
to be
erythrocytosis
of the
electrophoresis
this
Thus,
serum,
anoxic
of
( a)
with
and
in cyst
fluid
cyst
and
(d)
a
electrophoretic
and
IN
patients
anoxia,
a renal
activity
and
characteristics
from
due
to
to (c)
cerebellar
hemangioblastomas
by these
lesions.
factores
tu-
erythropoietin.11’34
molecular
serum
determinants.
SUMMARIO
de
in-
these
stimulating
further
a2-globulin
anemic
pathogenesis
for
in one
that
are
for
factor
some
in the
biological
an
antigenic
in human
except
Androgenic
assay
the
also completely
neutralized
of urine
of anemic
patients.
common
factors
cyst
Therefore,
The
as
treatment
tumors44
assay
all of which
are known
give in fact negative
same,
or nearly
or anoxia,
the
compared
( )
b
with
erythremia
mucoproteins
places
to
or anoxic
SUMMARY
have
migrate
used.
requires
found
to
pheo-
components
The
which
mouse
polycythemia
be
possible
hormone,
animal,
that
tissues
cysts,
all
patients,
systems
show
tumors
adrenocortical
erythropoiesis
assay
four
polycythemia.
is entirely
and thyroid
the experimental
the
occurs
where
of tumor
that
logical
in anemic
these
It
stimulate
erythropoiesis
and
found
destroyed
blastoma
done.4244
studies
hemangioblastoma.
materials
from
or polycythemic
produce
with
was
with
or serum
by the
rat
tumors
patients
terials
associated
by some tumors
is the
in response
to anemia
anemia
from
virilizing
which
present
stimulating
aplastic
fossa.
and
when
detected
the
in
In
not
posterior
steroid
hormones,
erythropoiesis
in
although
results
Thus,
negative
may
This
suggests
of erythropoietin
the
been
that
renal
)
fibromas43
tissue
substances
adrenocortical
to stimulate
and
have
in tumor
has
anoxia.38’39’4#{176} Consider-
hypernephroma,
syndrome.41
of production
WALDMANN
of erythropoietin
than
the kidney,
assays
hemangioblastomas
uterine
erythropoietin
or
erythropoietin
( i.e.,
positive
cerebellar
Hepatomas,542’44
anemia
It is of interest
in which
been
and
chromocytoma,
animals
unknown.
polycythemia
have
to
that
extra-renal
production
and anoxia.39’4#{176} The sites,
other
is produced
associated
or
in response
AND
quales
erythremia
characteristicas
se
trova
causate
molecular
in
per
le
seros
anoxia
de
e in
From www.bloodjournal.org by guest on June 14, 2017. For personal use only.
PROPERTIES
OF ERYTHROPOIESLS-STIMULATING
le liquido
biologic
un
in cata-un
globulina
indica
Le
de cystes
de patientes
un hemangioblastoma
(d )
renal,
activitate
facite
significa
que
ab patientes
esseva
etiam
un
extracto
anti
con
de
ha in commun
determinantes
le quales
le pathogenese
de
del
in le liquido
cystes
renal
erythrocytose
que
cerebellar
e hemangioblastomas
ille
pare
ab
un
anemic.
antigenic.
Assi
Isto
il pare
in sero
human
in hemangioblastoma
si non
es vidite
esser
per
patientes
es trovate
de cyste
es simile,
Isto
electrophoretic.
completemente
urina
le erythropoiese
e anoxic,
como
e trypsina.
mobilitate
neutralisate
un cyste
activitate
le
zonal
sialidase
simile
stimulante
anemic
per
con
(c )
a
que
in electrophorese
le mateniales
e in le liquido
poietina
migra
le tractamento
es mucoproteinas
in conilios,
le factores
bellar,
materiales
per
erythropoietinic
antisero,
747
con erythremia
secundari
cerebrellar.
Esseva
trovate
quatro
le materiales
que
que
del
e es destruite
02
FACTORS
identic.
Per
in association
causate
per
cereconsequente,
con
cystes
le secretion
renal
de
erythro-
lesiones.
ACKNOWLEDGMENTS
We wish to express
our
electrophoresis
experiments
excellent
technical
assistance.
appreciation
and to Miss
to Dr.
1)elores
John
L.
Houston
Fahey
for helpful
and Mrs.
Eleanor
advice
in the
Hull for their
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From www.bloodjournal.org by guest on June 14, 2017. For personal use only.
1964 24: 739-749
A Comparison of Some Physical and Chemical Properties of
Erythropoiesis-Stimulating Factors from Different Sources
WENDELL F. ROSSE and THOMAS A. WALDMANN
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