From www.bloodjournal.org by guest on June 17, 2017. For personal use only.
Acquisition
of Autophagic
Vacuoles
Physiological
By
Using
interference-contrast
erythrocyte
vacuoles
can
easily
dowing
Role
P.
CHm5T0PIIER
by Human
of the
HOLROYDE
AND
microscopy,
rapidly
and
noted
be
distinguished.
Because
effect
inherent
in
of the shathis
optical
bers
system
they
appear
as
surface
studied,
Spleen
H.
FRANK
in
all
GARDNER
splenectomized
They
were
rare
in
spleens.
Transfusion
tions.
Twenty
splenectomized
patients
with both normal
and abnormal
erythropoiesis
had 54.3 per cent vacuolated
cells
that these
inclusions
can
normal
mature
erythrocytes
as compared
to 2.1 per cent in a control
group
of normal
adults.
A splenectomized
they
lating
cyte
inclusions
and
morphologically
bin
F
degeneration.
vacuoles
of
Similar
inclusions
OLLOWING
like membrane
microscopy.1
fants
and
We have
splenectomized
with
These
cells
described
This
paper
presents
are,
indeed,
largely
inclusions
made
and
to
define
vacuoles
the
normally
patient.
from
the
all circulating
that
these
be
by
erythrocytes
viewed
with
when
large
with
that
illusions
beneath
nature
acquired
by
when
circuFurther,
circulation
suggests
that, in the
autophagic
vacuoles
process
numbers
evidence
be
with
show
in
absence
of
may occur
red cells.
It is
inclusions
would
removed
as
a physiological
a functioning
spleen.
were
occurred
in
the presence
optical
asplenic
normals
studies
of an intact
spleen
without
loss of red cell viability.
The
in situ in
presumed
were
reported
patients
low
an
cleared
evidence
a spleen,
hemoglo-
SPLENECTOMY,
indentations
sis as compared
spleens.2
previously
which
suggestive
in
are
the presence
measurable
patient
with
pure red cell aplasia,
whose
erythrocyte
mass
was
maintained
with
fresh
normal
blood,
had
44 per
cent
vacuolated
cells.
Electron
microscopy
thin
sections
showed
numerous
erythro-
subjects
and were
found
in greater
numin patients
with
young
cell popula-
tions.
intact
indenta-
Erythrocytes
of
numbers
of such
cells
in
both normal
and
abnormal
in older
children
circumstances
of vacuolated
the
and
seen
reflect
the
membrane.
inclusions
and
with
presence
Further,
their
crater(I-C)
newborn
in-
erythropoie-
adults
where
other
intraerythrocyte
indentations
which
the cell
these
appear
to develop
interference-contrast
with
intact
workers3’4
had
inclusions.
I-C
microscopy
of erythrocyte
an attempt
role
in
is
normal
physiology.
From
the
Hematology
Research
Laboratory,
Presbyterian-University
of Pennsylvania
Medical
Center
and the Department
of Medicine,
University
of Pennsylvania
School
of
Medicine,
Philadelphia.
Pa.
Submitted
January
29, 1970; revised
June 4, 1970;
accepted
June 9, 1970.
Supported
in parts by Grants
AM11188-03
and FR05610
from
the USPHS.
CHRISTOPHER
P. HOLROYDE,
MB.,
M.R.C.P.:
Fellow
in Hematology,
PresbyterianUniversity
of Pennsylvania
Medical
Center,
Philadelphia,
Pa. FRANK
H. GARDNER,
M.D.:
Professor
of Medicine,
Univorsity
of Pennsylvania
School
of Medicine;
Acting
Director
of
Medicine
and Director
of Hematology
Research,
Presbyterian-University
of Pennsylvania
Medical
Center,
Philadelphia,
Pa.
566
BLOOD,
VOL.
36,
No.
5
(NovEsdnEa),
1970
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HOLROYDE
GARDNER567
AND
MATERIALS
AND
METHODS
Patients
Control
subjects
indices,
normal.
In
at
1966,
bone
3 months
A.B.,
the
basis
of
59Fe
precursors.
of
over
mately
2-week
benefit.
Donor
1962.
intervals.
donors
at
10
was
healthy,
hematological
film
were
surgically
judged
documented
with
a marked
was
less
in
working,
and
had
normal
decrease
0.001
per
routine
volunteers
cent
clinical
traumatic
rupture
hematologic
with
and
approxi-
without
for
in
a period
at
1969
removed
red
count,
During
transfusions
August
normal
male
pure
platelet
date.
than
blood
spleen
and
selective
to
regularly
compatible
from
cell
idiopathic
a normal
healthy
suffering
white
performed
had
as
normal
days
normal,
male
apparently
blood
with
diagnosed
remains
count
fresh,
Splenectomy
were
was
condition
reticulocyte
a 25-year-old
apparently
was
blood
female,
with
whose
and
patients
reticulocytopenia,
Her
her
personnel
count
examination.
incorporation
maintained
D.F.,
He
Other
6 months
was
to
anemia,
red
laboratory
hospital
53-year-old
hemoglobin
cell
and
reticulocyte
were
prior
a
cent
mass
hospital
hematocrit,
marrow
cell
in
least
on
study
red
adult
subjects
patient
aplasia
8 per
of
of
hemoglobin,
Splenectomized
splenectomy
cell
consisted
including
indices.
intact
spleens.
Microscopy
I-C
microscopy.
Blood
phosphate-buffered
stored
for
up
to
Erythrocvtes
which
individually
any
as
red
communication
simply
times
in
x
the principle
the number
to
cell
20
palladium
estimate
appearing
of
cells
wet
to
either
were
of
with
no
of
the
Reichert
cells.
discrete
or
as
the
“pocked”
to be
in
least
500
cells
cell
were
cell.
be
In
referred
de-
on
a
to
its
previous
erythrocytes.
will
be
was
indentation
per
“pitted”
they
could
morphology.
A vacuolated
crater-like
or
potassium
cells
interference-contrast
At
number
misleading,
cent
alteration
of vacuolated
more
per
). Fixed
apparent
optics.5
or
1.0
milliosmoles
of Normarski
indentation
terms
volumes
290
using
one
described
these
20
7.4,
preparation
have
size
shows
vacuolated
12
temperature
a
Since
the
in the
text
cells.
electron
gluteraldehyde
buffered
room
on
presented
as
Scanning
at
in
(pH
as
these
to be
fixed
solution
viewed
regardless
work
immediately
is based
scanned
surface,
12
1 month
were
microscope
fined
was
gluteraldehyde
microscopy
solution
deionized
volumes
mm.
glass
alloy
and
cover
(SEM).
Cells were fixed in 0.5 per cent potassium
for 1 hour at room temperature.
They were then
distilled
water
and monolayer
films were prepared
slips.
viewed
Monolayers
with
the
were
Cambridge
coated
under
“Stereoscan”
Electron
microscopy
(E-M).
Cells
were
either
saline,
then fixed in 0.1 per cent glutaraldehyde
high
vacuum
electron
washed
three
with
microscope
times
in White’s
phosphatewashed
10
on clean
in
saline,6
gold-
(Mark
phosphate
or fixed
1).
buffered
directly
in 1.0
cent phosphate-buffered
glutaraldehyde
spinning
and was further
fixed with a solution
para-formaldehyde
in Na cacodylate
buffer.7
solution.
A red cell button
was produced
by
of 5 per cent glutaraldehyde
and 5 per cent
Postfixation
was
with 2.0 per cent Os04
in
buffer
for
Ui hours
at 2#{176}C.After
washing
in 0.5 per
cent uranyl
acetate
in
acetate
buffer,
cells were
successively
dehydrated
and embedded
in Epon
812.
sections
(60-80
m)
were cut with a Dupont
diamond
knife on a Sorvall
MT-2
per
s-collidine
veronal
Ultrathin
ultramicrotome,
and
stained
observed
with
Trans’,jsion
Patient
normal
cell
was
A.B.
indices.
A poo!ed
the
taken
transfused
obtained
acetate
and
lead
electron
to
was
recipient.
in
fate
of vacuolated
a healthy
with
1 unit
two
of
hydroxide,8’9
mounted
on
copper
grids
microscope.
of
51Cr-labeled
clearance
with
75
were
cells
blood
pCi
a normal
with
from
vacuolated
Na
51Cr
the
compatible
obtained
at
and
within
fresh
were
obtained
volunteer
of
with
blood
radioactivity
compatible
the
occasions
donor
labeled
Samples
hematocrit,
from
compare
on
samples
sample
the
changes
demonstrate
was
hypertransfused
Appropriate
within
measure
uranyl
7 (JeoLco)
Studies
donors.
half-life
with
a JEM
chromate
intervals
intact
splenectomized
cells
with
donor
blood
the
was
D.F.
radioactivity.
RBC
recpient
to
cells.
of blood
He
red
the
vacuolated
1 unit
four
normal
to assess
of
spleen.
from
ensure
from
number
recipient,
an
blood
to
(500
then
To
ml.)
rapidly
Samples
were
For
this
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568
ACQUISITION
OF
AUTOPHAGIC
VACUOLES
Fig.
1 .-Erythrocyte
morphology
viewed
with
the
interference-contrast
microscope.
Wet
preparation,
original
magnification
X 1000.
(A) normal
adult
cells;
(B)
cells
from
an adult
subject
with
“normal”
erythropoiesis,
splenectomized
for traumatic
rupture;
(C and D) cells from
splenectomized
subject
A.B. with
pure
red cell
aplasia.
study,
cells
vacuolated
were
counted
per
2000
incorporation
studies
were
performed
and
RBC
studies
were
performed
as
survival
RBC.
Routine
obtained
as
described
by
hematological
described
Read
et
by
data
Dacie
and
and
59Fe
Lewis.10
al.1t
RESULTS
Normal
with
I-C
erythrocytes
as crater-like
cent
SD)
±
small,
from
were
uncommon
from
a normal
trast,
20 splenectomized
had
54.3
vacuoles
diameter)
on
these
per
per
0.2
constituting
adult
cent
was
In
±
biconcave
appearing
were
normal
present
observed
to
with
possess
both
and
therefore
same
cells
more
shows
readily
that
identified.
the
were
membrane
than
and
to
cent
viewed
system
1.1 per
±
usually
single
1A).
Multiple
vacuoles
of all cells
and no cell
cent
cells,
optical
they
more
13.1 per cent vacuolated
(Fig.
1B-D).
They
tended
when
this
per
(Fig.
normal
discs
with
on 2.6
erythrocytes
to 0.8 p. diameter
less than
0.01 per
patients
cell.
uniform
vacuoles,
indentations,
of cells.
ranging
as
seen
Discrete
membrane
(mean
and
were
microscopy.
five.
In
abnormal
with
be
from
larger
Scanning
indentations
marked
con-
erythropoiesis
one
(0.2
electron
to over
p. to
1.5
20
p.
microscopy
seen
with
I-C
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HOLROYDE
AND
569
GARDNER
Fig.
2.-Scanning
electron
micro-
graph,
original
magnification
Erythrocyte
from splenectomized
A.B.
A
large
dentation
cavity
of
tions
microscopy
intact
large
in
are
fects,
however,
disorders
but
shown
( Fig.
membrane
observed
the
cell.
also
noted
Genuine
within
Two
in-
the
smaller
con-
indenta-
(arrows).
crater-like
membrane
de-
were
regularly
seen
in splenectomized
subjects
with
various
were
found
on less than
1.0 per cent of red cells. An example
is
2 ) . None
were
seen
in a control
group
of normal
adults
with
splenectomized
patient
vacuolated
cells.
other
splenectomized
vacuoles
inclusions
E-M
which
3 A-D.
The
structures
penheimer
as
a rare
etc.,
patients
are
mitochondrial
and
in all splenectomized
patients
with
vacuolated
than
persons
with
splenectomy.
illustrated
for
(Fig.
similar
studied
cell
cell
aplasia
had
blood
erythrocyte
none
to those
(Fig.
5),
populations
and,
nor
with
They
I-C
with
the
looked
where
The
were
per
cent
like,
and
and
The
microscopy.
surrounded
red
by
cell
envelope.
normal
density.
erythro-
Some
Fig.
“empty,”
other
erythrocyte
granules,
Papthe peripheral
of these
structures
(Fig.
4 A,B).
Apart
of these
were
described
being
found
in
reticulocytes
inclusions
44.0
of this patient
inclusions
bodies,
siderotic
time
to time
in
Examples
comparison
4 C),
erythrocytes.
spleens
Large
for,
Howell-Jolly
are seen
from
remnant
mature
in reticulocytes.
red
connected
stained
vacuoles
subjects
young
intact
demonstrably
bodies,
which
following
Inclusions
pure
to those
seen
studied
in detail.
neither
bodies,
other
with
and ranged
from material
which
resembled
smaller
collections
with
increased
electron
an amorphous
debris,
while
others
appeared
Heinz
particularly
from
not
inclusions
such
blood,
corresponding
A.B. were
was
They
were variable
cyte
cytoplasm
to
vacuoles
contained
A.B.,
thin sections
of the peripheral
patients
showed
numerous
apparently
from patient
a membrane
in
illusory.
are
12,000.
patient
spleens.
The
A.B.
measure
crater-like
is
X
seen
taken
from
in patient
A.B. were
noted
in greater
numbers
tended
described
to be
were
more
rare
circumstances,
occurred
predominantly
inclusions
as illustrated
(Fig.
3 C,D)
in
in these
vacuolated
noted
in this group.
\Vhen
patient
A.B. with
red cell
sions
with
fresh
normal
compatible
were
not
decreased
by
alterations
in
to base-line
observe
1
26 per
red
values
vacuoles
cent
cell
and
mass
aplasia
blood,
40 per cent
34 per
was
was hypertransfused
the number
of
(expected
cent
and
decrease
52 per
on two
vacuolated
calculated
cent
with
15 and 21 days, respectively,
(Fig.
6) demonstrating
could
develop
in normal
mature
erythrocytes
when
in
occacells
from
return
the
cir-
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570
ACQUISITION
Fig.
3.-Hypothetical
Splenectomized
surrounded
adjacent
patient
a double
by
erythrocyte
of
pure
cytoplasm.
included
material,
ternal
configuration
zone
shows
figures
sequence
A.B.
with
membrane
vacuolation
suggests
A
and
that
“in situ”
red cell
but appears
later
the
two
containing
material
resembling
increased
electron
density
with
imply
some
longevity
of these
AUTOPHAGIC
degradation
aplasia.
of
(A)
identical
inclusion,
presence
vacuoles
myelin
respect
structures.
OF
below,
inclusion
other
shows
(arrow)
respects
culating
within
D.F.
these
spleen,
(Fig.
normal
remnant
of
they
cells
T 3 of donor
erythropoiesis,”
had
cells were
transfused
slowly
7). Evidence
disappeared
for their
is derived
cells
the
The
ex-
of
A
figures
surrounds
this inclusion
to the surrounding
cytoplasm.
(C) A large
inclusion
shows
clear
which
Myelin
marked
is reminisvacuole
is
contains
a
material.
in an asplenic
patient.
The 51Cr
the recipient
was 27.5 days
(normal
with
“normal
vacuolated
is
to
condensation
of an amorphous
debris.
may have
coalesced.
(B)
condensation
and vacuolation
of constituents.
In this instance,
the debris
cent of degenerating
lipid material,
possibly
membrane.
A much
smaller
also noted
in this cell (arrow).
(D) Two large
vacuoles,
one of which
small
material.
inclusion
An
in
VACUOLES
from
from
subject
49.5 per cent
into a normal
from
continued
the
D.F.
T 3 of pooled,
26-32
days).
the
peripheral
circulation
minimal
in the
transfused
donor
Splenectomized
vacuolated
recipient
blood
loss
of radioactive
normal
recipient
cells.
\Vhen
with
an intact
over
in the recipient
48-72
hours
as apparently
label.
was
cells
donor
The
30 days.
t1Cr
From www.bloodjournal.org by guest on June 17, 2017. For personal use only.
HOLROYDE
AND
Fig.
4.-Other
patient
with
are seen
as coarse
inclusions,
spleen
and
longitudinal
electron-dense
of
A rare
with
acute
illustrated
sideroblastic
for
comparison.
anemia.
Iron-laden
and transverse
section
(M).
Siderotic
collections.
The cell is a reticulocyte
polyribosomes
patient
absence
(C)
erythrocyte
intact
in
cytoplasmic
mized
571
GARDNER
(PR).
a congenital
(B)
A
unstable
surrounding
membrane.
mitochondrial
remnant
well
formed
(Blood
from
Note
kindly
body
the
provided
splenectomized
patient
(SG)
appear
from
scattered
from
a
irregular
by
Alcoholic
mitochondria
granules
as seen
Heinz
hemoglobin.
(A)
splenecto-
structure
Dr.
and
Steven
Feig.)
A.B.
DIsCUSSIoN
E-M
Scanning
membrane
for the
That
they
effect
inherent
superior
appear
as
in
Heinz
phase
Koyama
that
I-C microscopy
vacuoles
of
erythrocyte
are
low
that
siderotic
erythrocyte
granules,
inclusions
etc.,
following
splenectomy.1315
The
pathology
as on splenectomy
et
al.3
structures
appearing
in
1962
noted
light
as
in large
measure
illusory
optical
density
within
the
structures
results
from
the peculiar
optics.12
In our hands,
I-C microscopy
optics
in allowing
small
erythrocyte
observed.
recognized
bodies,
peripheral
blood
much
on underlying
microscopy,
surface
Nomarski
to standard
easily
and rapidly
It has long been
bodies,
show
photographs
indentations
with
most
part,
represent
are
such
variably
and,
cell.
shadowing
clearly
was
vacuoles
to
be
as Howell-Jolly
noted
in
the
type and number
depend
as
per se. Using
standard
phase
and
dark
inclusions,
referred
From www.bloodjournal.org by guest on June 17, 2017. For personal use only.
572
ACQUISITION
to as “postsplenectomy
mized
subjects.
The
described
tamed
in
et
of
al.4
The
vacuoles
E-M
appearance
might
contain
Kent
et al.
membrane.
ing
al)normal
aplasia
had
Since
autologous
cell
maintained
lated
in
cells
with
could
an asplenic
provide
The
vacuolated
production
appearance
densation
fresh
normal
develop
blood
The
two
of
and
erythrocyte
vacuolation
the presence
imply
that they
have
demonstrated
splenectomized
only
contain
isoenzyme,
enzymes.19
tive
were
within
enzymes
almost
normally
may
never
persons,
as
apnear
empty.
II
be
seen
by
but
that
both
(endocytosis
all
similar
splenectomized
studies
bound,
were
in this
based
to
on
the enzymatic
phagocytic
cells
paper
the
E-M
degradation
with
poorly
workers
have
shown
occaautophagic
vacuoles
from
an ubiquitous
erythrocyte
significant
Since
spleens,
?)
A similar
to
vacuoles
vesicles
those
we
subjects
in the
have
studied
been
Further
described
and
were
the
that
also,
proposed
in
small
the
for
patient
in
slow
obligatory
but subsequently
form
larger
is
normal
many
vacuoles
morphologi-
that
an
proposed
noted
inclusions
they
are
from
namely,
work
reaction
degrada-
Conversely,
necessarily
to
additional
lysosomal
other
erythrocytes
explanation,
has
that
that
occurs.
stage
is not
this
by
large
vacuolated
it is presumed
in a few
reticulocytes.2#{176}
this phenomenon.
subjects
I and
form
independently
have
coalesced
mechanism
that
shared
is possible
vacuolation
alternative
to
It
enzyme
observed
an
suggested
a property
soluble.
represent
structures
of mitochondria
in dog
laboratory
to investigate
Inclusions
similar
certain
similarly
localized.
in subjects
with
suggest
It is possible
after
small
in
III
inclusions
5).
by
others.2#{176} In splenectomized
could
represent
an end
from
3 A and
events
Certainly,
other
activity
in some
phosphatase
is
may
regularly
cal sequence
progression
(Figs.
and
before
These
reported
vacuo-
circulating
from
this patient.
The
progressive
conmaterial
was morphologically
suggestive
fractionation
was
particle
were
noted
that
reaction
vacuoles
removed
vacuoles
‘em)tv
implied
product
within
these
vacuoles
may not necesautophagic
in nature.
Kornfeld
and Gregory,18
howthree
isoenzymes
of acid phosphatase
in erythrocytes
donors,
while
those
from
subjects
with
intact
spleens
I,
the
of
pure
red
interest.
cell mass
are
Enzymes
product
postulate
that
by a limitassociated
when
studies
is so.
sequence
appearing
hemoglobin
of
Cell
two.
enzyme
this
erythrocytes
hypertransfusion
developed
lysosomal
systems’6’17
sional
localized
acid
phosphatase
human
red
cells.4
Because
acid
enzyme,
normal
inclusions
of included
degeneration,
or autologous
workers
to
surrounded
predominantly
transfusions,
in previously
patient.
of hemoglobin
of isologous
sarily
ever,
from
from
splenectoto the vacuoles
finding
that
patient
A.B.
with
cells was therefore
of particular
approximated
zero
and
red
further
evidence
that this
have
proposed
a hypothetical
\Ve
of red cells
equivalent
lead
both
groups
of
degenerating
hemoglobin
proposed
that
they
are
erythropoiesis.
44 per cent
red
cell
with
was
in 50 per cent
are probably
VACUOLES
study
using
I-C microscopy
and close
agreement
was obper cent
of vacuolated
cells
were
found
in a comparable
twenty
splenectomized
patients.
The
postsplenectomy
vacuole
can
be correlated
with
the “autophagic
vacuole”
described
later
by Kent
probably
such
vacuoles,”
inclusions
AUTOPHAGIC
this
54.3
since
group
light
OF
the
A.B.
fuse
vacuoles
elimination
progress
in greater
step.
in
were
numbers
this
found
in
From www.bloodjournal.org by guest on June 17, 2017. For personal use only.
HOLROYDE
Fig.
5.-Red
tomized
empty”
An
appear
with
about
vesicle
young
cent
cell
lack
by
therefore,
selectively
marked
in
eliminated
from
viability.26
donor
were
In
intact
spleen
in 48-72
T % normal,
dogs,
was
three
in
for
a larger
this
study
hours.
the
reticulocytes.
process
would
by Koyama
They
supravital
cell
by
loss
on
the
and
appear
to be
et al. in 196422
animals23
other
without
cells
from
of a normal
of radioactivity
a
and,
hand,
are
altering
the
a splenecto-
recipient
with
minimal
and
was
nonlethal.
though
iron
a function
inclusions
of Heinz
bodies
The process
is
the
spleen
vacuolated
do
removed
by
for removing
in laboratory
circulation
Since
bodies
as Howell-Jolly
bodies,
from
their
presence
in
Sequestration
of the spleen.
granules,
51Cr-labeled
from
sized
preformed
inclusions
Romanovsky,
destruction
red
medium
They
appear
to represent
per Se. The presence
of these
Siderotic
the
splenec-
a
spherical
particularly
red cells.22’
to be a function
man.
cleared
the
noted
and
into
structures
such
etc.,
as is evident
erythrocyte
presumably
mized
erythropoiesis”
(M)
implies
that
they
would
normally
be
unique
architecture
is well
adapted
whose
cell’s
“normal
noted.
of affinity
or inclusion-containing
shown
by Rifkind24
accompanied
51Cr
using
also
populations,
of cells,
splenectomy
spleen
with
containing
are
their
distinctive
morphology.
as well as of splenectomy
following
functioning
damaged
has been
(AV)
(Vi)
other
well
recognized
siderotic
granules,
50 per
and
age
a patient
vacuole
“empty”
not represent
Heinz
bodies,
stains,
of cell
from
previously.
A mitochondrion
to be discharging
their
contents
years
autophagic
a small
patients
inclusions
4
vacuole
vacuole.
and
cell
TTP
for
573
GARDNER
AND
A similar
E-M
sections
an
the
finding,
of the
Tx
50
AB53
Fig.
40
30
VACuOLATED
CELLS
#{176}#{176}
20
40
HCT
0
30
#{176}‘
0-0
o
io
zo
DAYS
IL)
‘O
DO
of
6.-Hypertransfusion
splenectomized
with
red
cell
aplasia.
of vacuolated
cells
return
about
to
20
patient
base-line
days.
.
A:B.
Dilution
occurs
values
with
in
From www.bloodjournal.org by guest on June 17, 2017. For personal use only.
ACQUISITION
574
OF
AUTOPHAGIC
VACUOLES
00
00
80
80
Fig.
60
60-f
7.-Clearance
of
51Cr-labeled
transfused
vacuolated
VAUOLATED
cells
from
the
circulation
cipient
of a normal
rewith
an
intact
Note
the minimal
of radioactive
label.
spleen.
20
loss
30
0
70
50
HOURS
spleen
achieved.
failed
to demonstrate
It is conceivable
surface
and
that
Although
the
much
dated,
the
form
vacuoles
proposed
sequent
splenic
milieu
information
evidence
and
the
precise
manner
vacuoles
extrude
their
that
suggests
that
the
facilitates
such
concerning
that
may
normal
are
remains
erythrocytes
removed
reflect
removal
the red
via
was
cell
a process.
mechanisms
mature
majority
that such inclusions
on erythrocyte
aging.
by which
contents
by
the
a degradative
to
may
be
eluci-
continually
normal
spleen.
It is
cellular
process
con-
ACKNOWLEDGMENT
The authors
gratefully
acknowledge
the expert
technical
assistance
of Mrs.
Judith
Burden
and
Mr. Josef \Veibel.
They
are indebted
to Dr. Alastair
McKee
of the Pioneering
Research
Laboratory,
Experimental
Station,
E. I. duPont
du Nemours
and Co., Wilmington,
Del., for
his skill in preparing
and viewing
cells with the “Stereoscan”
electron
mcroscope
and to Dr.
Max A. Listgarten
of the Center
of Oral Health,
University
of Pennsylvania,
for his advice
concerning
the preparation
of electron
micrographs.
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From www.bloodjournal.org by guest on June 17, 2017. For personal use only.
1970 36: 566-575
Acquisition of Autophagic Vacuoles by Human Erythrocytes Physiological
Role of the Spleen
CHRISTOPHER P. HOLROYDE and FRANK H. GARDNER
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