From www.bloodjournal.org by guest on June 17, 2017. For personal use only.
Fibrillar
Bundles
from
Leukemic
By
D
URING
project,
nucleus
of blood
cytes
obtained
apparently
this
from
leukemic
patients,
the
the
number
to
the
lymphostructures
lymphocytes.
study
of
the
The
morphologic
normal
these
structures
in their
blood
lymphocytes,
significance
of their
presence
in the nucleus.
with
several
of
intranuclear
human
(1)
:
another
in the
Although
cytoplasm
such
in
was
on
bundles
donor.
in
observed
to determine
TONAKI
a normal
formations
therefore,
(2 )
fibrils,
Patients
INVESTIGATION
made
of fibrillar
was
previously
investigation,
these
individuals
understand
and
been
Lymphocytes
HIROSHI
AND
fibrillar
normal
never
STEFANI
obtained
reported
of Blood
Nonleukemic
MICROSCOPIC
observation
lymphocytes
from
of
of
S.
STEFANO
have
have
aspect
and
ELECTRON
the incidental
investigators14
purpose
in the Nucleus
and
leukemic
(3 )
and
to try
to
METHODS
Lymphocyte
Blood
Blood
samples
defined
by
the
also
vere
equal
To
following
and
inclil)ated
adhere
of
1 per
The
This
suspensions
were
buffer
) added
of
the
were
then
37#{176}C. During
per
cent
of
was
fixed
were
introduced
this
procedure,
the
nucleated
collected
from
( (hInted
to adjust
fixative
osmium
cells
at
final
for
as
addition,
as
controls)
patient
and
the
supernatant
in
medium
which
sedi-
consisted
serum.
lymphocytes
were
each
leukocytes
the
in
human
In
( considered
from
37#{176}C. The
leukemia
disease.
diseases
obtained
resuspended
fibrils,
lymphocytic
Hodgkin’s
concentrated
into
the
by
in
the
electron
means
flat-bottomed
granulocytes
cells
the
a
of
T60
and
flask
monocytes
supernatant
are
microscopic
seen
study.
Preparations
in \Iillonig
( wt./vol.
cent
these
90-99
at
chronic
with
were
compatible
suspensions
supernatant
Microscopy
p11
for
mm.
and
hematologic
mm.
and
The
30
glass
lymphocytes.
The
The
search
no
blood
and
(Difco)
with
patients
of
30-60
for
patients
eight
with
washed,
TC199
for
the
Electron
hyde
NaCI
out
the
16
and
milliliters
procedure:
to
be
patients
centrifuged,
facilitate
the
30
carried
parts
from
Gunz,5
Twenty
was
plasma
obtained
and
from
studied.
mentation
to
were
Dameshek
lymphocytes
were
of
Separation
was
tetnoxide
dehydrated
the
adjusted
in
16
1 : 1 with
to
the
osmolality
to
Veronal
through
hr
7.4.
the
The
with
fixative
suspensions
( pH
of
days
distilled
of
buffer
series
several
proper
7.4
increasing
1.5
)
to
a value
were
and
per
water
340
concentrations
cent
with
of
then
mOsM
glutaralde-
0. 13
per
340
postfixed
) for
of
one
alcohol
cent
mOsM.
with
hour.
and
From
tile Electron
Microscopti
and Radiobiology
Research
Sections,
Veterans
Adnlini.stration
Hospital,
Hines,
Ill.,
an(l
Department
of Radiology,
Tile
Chicago
Medical
School,
Chicago,
Ill.
First -ubmitted
July 2, 1969; accepted
for J)ublication
October
3, 1969.
Tiii-s work Was Sll;)porte(l
ill
part
by tile Leukemia
Research
Foundation,
Inc., Chicago,
Illinois.
STEFANO
S. STEFANI,
NI .D. : Chief,
Therapeutic
Radiology
Service,
Veterans
Adni inistration Hospital,
Hines,
Ill., and Clinical
Professor
of Radiology,
The Chicago
Medical
School,
Chicago,
Ill. HIROSHI
TONAKI,
B.S. : Electron
Microscopist,
General
Medical
and
Surgical
Research
BLooD,
Service,
VOL.
Veterans
35, No.
Administration
2
(F’LBRUAJSY),
Hospital,
1970
Hines,
Ill.
243
From www.bloodjournal.org by guest on June 17, 2017. For personal use only.
STEFANI
244
Fig.
present
Fig.
shaped
nucleus
1.-Normal
in nucleus
2.-Normal
lymphocyte.
(N).
lymphocyte.
arrangement
close
(N), mitochondrion
Pairs
of
Intranuclear
to nuclear
(M).
arrows
bundle
membrane
indicate
borders
of fibrils
(NM).
of fibril
(arrows)
Cytoplasm
AND
shown
marked
TONAKI
bundles
in Y-
(CY),
From www.bloodjournal.org by guest on June 17, 2017. For personal use only.
FIBRILLAR
BUNDLES
IN
THE
NUCLEUS
OF
BLOOD
245
LYMPHOCYTES
.‘
Fig.
3.-Normal
different
nuclear
propylene
than
lymphocyte.
Several
intranuclear
bundles
angles
and indicated
by arrows.
Nucleus
membrane
(NM),
mitochondnia
(M).
oxide,
1000
and
sections
finally
of
embedded
lymphocytes
in
were
araldite
studied
denoted
about
17,000
fibrils
(N),
( Fluka
mixture
at
of
by
sectioned
cytoplasm
) . In each
at
(CY),
case
more
magnifications.
RESULTS
Bundles
of fibrils
were
observed
in the
nuclei
of
blood
lymphocytes
of
15
of 30 nonleukemic
patients
studied.
These
fibrils
interchromatin
space
and were
arranged
in a parallel
were
always
found
in the
fashion
forming
a bundle
of
( Fig.
variable
length
bundles
were
peared
to have
and
seen
about
to form
a beaded
200
m
a Y-shaped
substructure
.
in
diameter
arrangement
and
when
1
(Fig.
seen
)
.
2).
in cross
In
one
The
instance,
fibrils
section
aphad
a
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246
STEFAXI
Fig.
4.-Normal
bundles
of
nucleus.
section
single
or
one
certain
could
bodies
cytoplasmic
given
sample
served
both
definite
nuclear
patients
appeared
occurred
proof
could
be
(arrows)
contain
found
any
angles
in
lumen.
were
the
In
observed
represented
separate
entire
interchro-
were
samples
only
often
be
than
out
all
of
samples
often
together
established
only
in
of
70-100
fibrils.
the
in the
and
concerning
the
they
intranuclear
same
and
small
On
bundles;
than
firmly
5).
studied,
intranuclear
more
found
found
be
structures
4 and
one
direction
not
these
bundles
in
in the
could
( Figs.
demonstrated
seen
times
extending
contact
between
intranuclear
always
seen
direct
in a sample
were
10-20
not
to
they
connection
these
more
were
seem
throughout
although
be
detectable
bundles
approximately
although
in
fibrils
at different
\Vhether
could
),
2
observed
that
contained
hand,
bundles
could
when
estimated
not
cut
winding
detectable
or nucleoli
of
(NB).
established.
( Fig.
no
structures
\Ve
sections
In
these
Further,
These
be
envelope
established.
nuclear
bundle
not
instances
nuclear
and
the
continuous
material,
of the
three
did
70
3 ) several
a
A
bundle
body
( Fig.
bundles
No
nuclear
approximately
matin
bers.
near
of
within
In
Intranuclear
)
instances
diameter
some
lymphocyte.
(IC1
material
TONAKI
,
#{149}.
interchroniatin
AND
nuin-
cellular
the
other
for
any
were
ob-
bundles,
section.
significance
of
these
bundles.
spite
of
of
extensive
16 patients
with
similar
searching,
with
Hodgkin’s
to those
chronic
disease.
found
these
fibrillar
lymphocytic
The
fibrils
in normal
bundles
leukemia,
found
patients.
in
were
found
and
in one
of eight
lymphoma
cases
the
in
only
From www.bloodjournal.org by guest on June 17, 2017. For personal use only.
FIBRILLAR
BUNDLES
Fig.
IN
5.-Normal
THE
NUCLEUS
OF
lymphocyte.
interchromatin
material
BLOOD
LYMPHOCYTES
Intranuclear
(1CM)
near
bundle
nucleolus
247
of
(NC)
fibrils
(arrows)
surrounded
found
by
in
chromatin
material.
DIscUSSIoN
The
observation
of these
structures
in
tients
without
hematologic
disease
contrasts
of
cytes
13
of
eight
patients
in incidence
16
patients
with
is the
stage
of maturation
he demonstrated.
In
that
spite
of
they
are
as
difficult
In
the
similar
lined
our
trate
even
segment
unless
no
proof
membrane
structures
the
membrane.14
during
fibrils
small
identify
study,
nuclear
low
are
Similar
of
incidence
rarely
in
the
these
of
of
the
cytoplasm
contact
findings
and
to
sections
are
because
of the
have
human
these
also
been
et al.6 reported
to the nuclear
merge
were
in
repeatedly
normal
appear
they
structures
and
the
and
described
have
outer
observed
in a few
appeared
chromatin.
of some
are
section.
have
and
also, that
membrane
with
the nuclear
strongly
suggestive
(b )
these
lymphocytes
fibrils
of
they
)
to
impression
longitudinal
investigators
of
remains
and
their
between
Several
between
structure
: (a
seven
difference
a different
it is our
nuclear
region,
cut
connection
made.
and
this
or
state,
bundles,
interchromatin
a direct
be
the
15 of 30 pain the lympho-
leukemia,
Whether
of population
however,
bundles
of
of a pathologic
of
observed,
this investigation.
Parker
arranged
perpendicularly
through
a defect
some
of our own
or
component
could
in the
frequency
lymphocytic
lymphocytes,
unusual
They
a
to
an
chronic
lymphocytes
their
absence
disease
investigated.
of a different
type
the
apparent
not
lymphocytes.
usually
of
the
with
Hodgkin’s
expression
blood
with
undernuclear
by
instances
to pene-
Although
connection
us
From www.bloodjournal.org by guest on June 17, 2017. For personal use only.
248
STEFANI
between
the
nuclear
and
cytoplasmic
nection.
These
bundles
are
pear
to have any connection
these
bundles
lymphocytes
Similar
do not resemble
by Tokuyasu
et
intranuclear
ing
into
the
The
observed
Bundles
and
and
pore
by
mice
been
found
in vitro
by
a definite
Bernhard
“viral
zone
found
have
in the
been
nucleoli
described
in
blood
The
in human
chick
sympaauthors,
extend-
the
from
Syrian
could
human
hepatocytes
et al.’2 detected
of hamster
cytoplasm
hamsters.
serve
as
of fibrils
resemble
former
cells,
and
and in the
In
an
addition,
also
be
the
of
those
a
viral
that
etiology
and
by
viral
noted
as Adenovirus
be considered
described
of
infected
Type-
they
by us in normal
expression
cells
also
infected
bundles
of
cells,
both
of Adenovirus
indicator
described
morphologically
could
at times
unknown.
by Rowe
nuclei
of thymic
Filaments
have
presented
the same
antigenic
character
suggest
that the bundles
of fibrils
could
bundles
lymphocytes
ap-
et al.5 These
envelope
and
in the
virus.”
in the nuclei
of
Similarly,
Kalnins
amnion
Type
12,
and
cells.
Granbolulan,1#{176}
“mouse
thymic
with
cells
of fibrils
authors
footprints”
neoplastic
Whether
and
infected
neoplastic
bundles
12. The
con-
fibrillar
bundles
by Patrizi
et al.’1
herpes
simplex
virus.
12-induced
be
show
area.
fibrils
in nuclei
of human
in vitro
with
Adenovirus
as
could
interchromatin
space
and do not
bodies
or nucleoli.
Morphologically,
nature
and
the function
of nuclear
fibrils,
as well
as those
in the cytoplasm
of the same
lymphocyte,
are
as yet
of fibrils
associated
with
virus
particles
have
been
described
newborn
these
Type
none
TONAKI
in the
nuclear
with
deuterium
by Masurovsky
approaching
close
to the nuclear
nuclear
Capps,9
of
the
al.7
fibrillar
thetic
neurons
treated
too, noted
some
fibrils
fibrils,
located
with
AND
in
leukemic
Kalnins
infection
et
al.
remains
to
represent
a
demonstrated.
Another
possible
paracrystalline
move
no
from
in
the
are
nucleus
structures
the
carried
of this
out
cytoplasm
study
However,
is that
or
our
of these
laboratory
which
may
We
could
versa.
cytoplasmic
studies
with
)
RNA
vice
autoradiographic
in
they
( possibly
material
to
or autoradiographic
literature.
being
composition
these
of nuclear
the
histochemical
reported
uridine
of
arrangement
eventually
find
explanation
the
bundles
using
of
aim
tritiated
testing
the
material.
SUMMARY
Fibrillar
of
30
bundles
normal
lymphocytic
Nuclear
bundles
numbers.
No
could
found
individuals.
chronic
fibrils
were
be
Similar
leukemia,
when
points
in
of
the
nuclei
fibrils
and
were
in one
observed
in
continuity
of
a
found
of eight
sample
between
blood
the
lymphocytes
in
cases
three
of
16
of Hodgkin’s
were
found
nuclear
and
15
cases
of
disease.
only
the
from
in
small
cytoplasmic
found.
ACKNOWLEDGMENT
We are
assistance.
indebted
to Mr.
Carl
Kirsh
and
Mr.
Joseph
Kompare
for
their
skillful
technical
From www.bloodjournal.org by guest on June 17, 2017. For personal use only.
FIBRILLAR
BUNDLES
IN THE
NUCLEUS
OF
BLOOD
249
LYMPHOCYTES
REFERENCES
1.
ture
J.
D. :
Zucker-Franklin,
of cells
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C.,
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and
Y.,
9:325,
F.,
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Jr. :
of lymphocytes
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Anderson,
D.
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ultrastruc-
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Epstein,
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normal
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The
thoracic
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Transformation
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Ultrastructure
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human
Suppl.
4. Wakasa,
H., Parker,
J. W., and Lukes,
R. J. : Cytoplasmic
filamentous
structures
in
activated
lymphocytes
: an electron
microscopy
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New
York,
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5. Dameshek,
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1964,
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2).
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and
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F. : Leu& Stratton,
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to growth
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8. Masurovsky,
E. B., Benitez,
H. H.,
and
Murray,
M. R. : Unusual
intranuclear
structures
in chick
sympathetic
neurons.
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Electron
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Chicago,
Ill., August-September
1967,
p. 188.
9. Rowe,
\V. P., and Capps,
W. I. : A
new
mouse
virus
causing
necrosis
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thymus
in newborn
mice. J. Exp. Med. 113:
831, 1961.
10. Bernhard,
W., and
Granboulan,
N.:
Morphology
of
oncogenic
and
nononcogenic
mouse
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Ciba Foundation
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of Murine
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Brown,
1962, p. 6.
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Reed,
plex
Patrizi,
C. A.
virus
newborn.
G.,
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of
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7. Tokuyasu,
K., Madden,
Zeldis,
L. J.: Fine
structural
interphase
nuclei
of lymphocytes
cells and in Adenovirus-12-infected
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hamster
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1967.
S. C.,
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V. I., Stich,
H. F., Gregory,
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anti-
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1970 35: 243-249
Fibrillar Bundles in the Nucleus of Blood Lymphocytes from Leukemic and
Nonleukemic Patients
STEFANO S. STEFANI and HIROSHI TONAKI
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