Levels in Stroke Patients of CSF Astroprotein,
an Astrocyte-Specific Cerebroprotein
T O R U HAYAKAWA, M.D.,
YUKITAKA U S H I O , M.D.,
N O R I O ARITA, M.D.,
TAKESADA M O R I ,
TOSHIKI YOSHIMINE,
YASUTAKA M A E D A , M.D.,
KEIJI SHIMIZU,
A N D A K I O M Y O G A , M.
685
M.D.,
M.D.,
M.D.,
SC.
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SUMMARY Astroprotein (an astrocyte-specific cerebroprotein) levels in cerebrospinal fluid (CSF) were
determined by radioimmunoassay in 47 stroke patients. (Astroprotein is immunologically identical to glial
fibrillary acidic protein.)
Astroprotein levels in CSF increased markedly in acute cases of intracerebral hemorrhage and slightly to
moderately in some acute cases of subarachnoid hemorrhage and cerebral infarction. In intracerebral
hemorrhage, CSF astroprotein levels in the acute stage of the ictus reflected the size of the lesion and were used
to estimate the clinical outcome. In subarachnoid hemorrhage and cerebral infarction, CSF astroprotein levels
were related to the general neurological state.
Evidence obtained indicated that fundamentally different destructive and/or degenerative processes in the
brain may be involved in intracerebral hemorrhage, subarachnoid hemorrhage and cerebral infarction and that
determination of CSF astroprotein may have clinical significance in stroke patients.
Stroke Vol 10, No 6, 1979
INVESTIGATION of brain-specific proteins has advanced rapidly in recent years. Astroprotein designated by Mori1 is the major component of Tay-Sachs
disease protein detected by Bogoch2 and is specific to
astrocytes and astrocytoma cells.36 Glial fibrillary
acidic protein (GFAP), extracted later by Eng et al.,6> 7
was immunologically proved to be identical with
astroprotein.8
It was reported that astroprotein levels in
cerebrospinal fluid (CSF) increased in patients with
glioma and it was suggested that the existence of a
destructive process in the brain might cause the increase of astroprotein in CSF.9> 10
The purpose of the present paper is to analyze CSF
astroprotein levels in stroke patients and study their
significance.
most of the patients (35) the first CSF samples were
obtained by lumbar puncture in the acute stage within
4 days after the ictus without neurosurgical procedures. From 14 patients, CSF samples were obtained on several occasions during the course of their
diseases.
The CSF samples were stored at -60°C until
assayed.
Methods of purification and radioimmunoassay of
astroprotein have been described elsewhere10 and only
a brief schematic outline is given in figs. 1 and 2.
Current radioimmunoassay is able to detect astroprotein in the range of 5-1000 ng/ml.
Astroprotein concentration was assayed in all CSF
samples and total protein level was simultaneously
determined in 36 of the samples.
Materials and Methods
Results
Ninety-four samples of CSF were collected from 47
patients in acute or subacute stages of stroke; intracerebral hemorrhage (ICH) 22 patients, subarachnoid hemorrhage (SAH) 11 patients, and
cerebral infarction (INF) 14 patients. Control CSF
samples were obtained from 8 patients without clinical
and laboratory evidence of intracranial diseases. From
Astroprotein concentrations in CSF of acute stage
stroke patients are presented infig.3. The mean value
of CSF astroprotein levels of the control patients was
10.0 ± 1.07 (SE) ng/ml. Regarding the values above
1000 ng/ml as abnormal, the mean values of astroprotein levels in CSF obtained from acute stage stroke
patients was 772.4 ± 90.14 ng/ml in ICH,
187.8 ± 109.05 ng/ml in SAH and 107.8 ± 57.77
ng/ml in INF. The astroprotein levels were markedly
and constantly high in the ICH group. In some of the
patients with SAH or INF, CSF astroprotein levels
also increased but only slightly or moderately. CSF
astroprotein levels of patients in acute stages of SAH
or INF did not exceed 1000 ng/ml except for a case of
SAH in critical condition. Statistical analysis by
Student's /-test disclosed significant difference
between the CSF astroprotein levels of ICH and SAH
(p < 0.001), the levels of ICH and INF (p < 0.0005),
and the levels of ICH and control (p < 0.0005).
From the Department of Neurosurgery, The Center for Adult
Diseases, Osaka, Nakamichi, Higashinari-ku, Osaka 537 (Drs.
Hayakawa and Yoshimine); Departments of Neurosurgery,
Surgery and Traumatology, Osaka University Medical School,
Fukushima, Fukushima-ku, Osaka 553 (Drs. Ushio, Mori and
Shimizu); Division of Cerebrovascular Diseases, Hanwa Hospital,
Karitacho, Sumiyoshi-ku, Osaka 558 (Drs. Arita and Maeda); and
Dainabot Radioisotope Laboratories, Toranomon, Minato-ku,
Tokyo 105, Japan (Mr. Myoga).
Reprints: Dr. Hayakawa.
This study was supported in part by a grant from the Research
Foundation for Cancer and Cardiovascular Diseases, Japan.
686
STROKE
glioma
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1979
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VOL 10, No 6, NOVEMBER-DECEMBER
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FIGURE
1. Procedures for purification of astroprotein.
ICH
Figure 4 presents the relationship between CSF
astroprotein levels in acute stage ICH and the size and
location of hematoma with or without rupture into the
ventricles. There is a tendency for patients with larger
hematomas or a hematoma ruptured into the ventricles to show higher levels of astroprotein in CSF.
Figure 5 shows the changes of CSF astroprotein
levels following the ictus when surgery is used. For a
few days after the onset, the astroprotein levels were
usually remarkably high, and then decreased
gradually. The decrease of astroprotein in CSF was
slower in patients treated by neurosurgical procedures, such as evacuation of hematoma, than in
those treated conservatively.
SAH
Figure 6 presents the relationship between CSF
astroprotein levels in patients with SAH and the
12S
I-AP
standard or sample
anti-AP (X10.000)
1 st incubation at 4°C for 4 8 " •72hrs
1 % NRS
anti-R-yG (X15)
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FIGURE
ICH
SAH
INF
patients. ICH: intracerebral hemorrhage. SAH: subarachnoid hemorrhage. INF: cerebral infarction.
clinical grade by Hunt and Kosnik.11 The patients with
higher grade showed higher levels of astroprotein in
CSF.
INF
Figure 7 presents the relationship between CSF
astroprotein levels and the levels of consciousness in
the INF patients. No patients in this series was in
coma at the time the first CSF specimen was collected.
The patients with slighter consciousness disturbance
showed lower levels of CSF astroprotein except for a
patient with brain-stem infarct, in which CSF
astroprotein on the 2nd day after ictus was very high.
In this patient, CSF astroprotein became higher on
the 3rd day and the infarct proved by CT scan to have
become hemorrhagic.
Clinical Outcome
IOO//J0
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2nd incubation at 4°C for 20hrs
centrifuge
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count
protein.
Control
FIGURE 3. CSF astroprotein levels of acute stage stroke
2. Procedures for radioimmunoassay of astro-
Figure 8 shows the relationship between CSF
astroprotein levels in acute stroke and clinical outcomes. In ICH, the clinical outcome of the patients
with lower astroprotein levels in CSF was better than
those with higher astroprotein levels. No definite
relationship was observed between CSF astroprotein
levels and clinical outcome in the SAH and INF
groups.
CSF ASTROPROTEIN LEVELS IN STROKE/Hayakawa
ICH
ICH
© ©
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687
et al.
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after ventricle drainage
after evacuation of
hematoma, etc.
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The changes of CSF astroprotein levels follow-
ing the ictus of ICH with reference to surgical procedures.
SAH
10c
Putaminal
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• small
® large
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FIGURE 4.
Thalamic Subcortical Cerebellar Brain stem
hematoma
hematoma
hematoma
hematoma
with rupture into ventricle
without rupture into ventricle
with rupture into ventricle
without rupture into ventricle
The relationship between CSF
astroprotein
levels in acute stage ICH and the size and location of
hematoma with or without rupture into the ventricles.
E
c
a
Z IO
UJ
O
CSF Total Protein
Figure 9 presents the relationship between astroprotein and total protein levels in CSF. There was
poor correlation between the 2 levels for all 3: ICH,
SAH and INF.
Discussion
Astroprotein, 1 which is immunologically identical
to glial fibrillary acidic protein (GFAP), 8 is the first
protein found to be specific for astrocytes and is
regarded as one of the components of the glial
filament,1-6> 8 Immunofluorescence studies using antiserum to astroprotein demonstrated that the specific
fluorescence was distributed in the cytoplasm of
fibrillary astrocytes in the normal brain, reactive
astrocytes and astrocytoma cells.1' *•6 Recently, this
protein has been successfully purified and radioimmunoassay with double antibodies has been developed
by Mori et al. 9 ' 10
<r
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a.
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10°
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1 2
3
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GRADING
by Hunt & Kosnik (1973)
FIGURE 6.
The relationship between CSF
levels and the clinical grades in SA H patients.
astroprotein
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FIGURE 7. 7/ie relationship between CSF astroprotein
levels and the levels of consciousness in INF cases.
Investigation of CSF astroprotein levels in patients
with brain tumors indicated that CSF astroprotein increased in patients with gliomas, especially malignant
gliomas.9-10 It seems likely that the mechanism of increased astroprotein in CSF might be connected to the
existence and degree of degenerative and/or destructive processes in the brain.
The present study demonstrates that CSF astroprotein increases in acute stages of stroke, especially in
ICH. In ICH, the fact that CSF astroprotein levels
were higher in patients with larger hematoma would
support the view that the increase of astroprotein in
CSF may be concerned with destructive processes in
the brain. In contrast to SAH and INF, the CSF
astroprotein levels in ICH increased remarkably soon
after the ictus. This may be due to differences of
destructive mechanisms of brain tissue among the 3
types of stroke. ICH may cause more rapid and more
serious destruction of brain tissue.
In SAH and INF, it was clear that the astroprotein
levels in CSF were related to the patients' neurological
state, i.e. clinical grade or consciousness level. The
general neurological condition may be considered a
reflection of the degree of destructive and/or degenerative processes in the brain.
10°
1
able to do active work
able to care for self
able to live at home with assistance
unable to care for self
dead
• ICH
o SAH
X INF
(
) : dead by complications
The relationship between CSF astroprotein
levels in acute stroke cases and clinical outcomes.
FIGURE 8.
There was poor correlation between CSF astroprotein levels and the prognosis in SAH and INF, though
there was good correlation in ICH. The mechanism
behind this difference remains obscure. In most of the
SAH cases, intracranial aneurysms were found and
treated neurosurgically. CSF was sampled within a
few days of the onset of ictus. Possibly re-rupture of the
aneurysms and late vasospasm, which happened after
CSF sampling, affected the clinical outcome. Furthermore, clinical outcome may have been directly influenced by surgical procedures which followed. In the
INF patients, the degenerative processes may be slow
and fluctuating. The timing of CSF sampling should
be further studied to clarify the relationship between
CSF astroprotein levels and clinical outcome in SAH
and INF.
There are numerous reports on changes in CSF i.e.
CSF ASTROPROTEIN LEVELS IN STROKE/Hayakawa el al.
•
o
X
aio 3
689
Acknowledgment
1 CH
SAH
1 NF
We wish to thank the Department of Neurosurgery, the Center
for Adult Diseases, Osaka, Departments of Neurosurgery, Surgery
and Traumatology, Osaka University Medical School and Division
of Cerebrovascular Diseases, Hanwa Hospital, for providing
samples of CSF. Grateful acknowledgment is made to Prof. H.
Mogami, Dr. K. Kamikawa, Dr. T. Sugimoto, Dr. M. Taneda and
Dr. T. Irino for their constant interest and valuable suggestions.
References
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ioc
50
100
I5OS
CSF TOTAL PROTEIN (mg/dl)
FIGURE 9. The relationship between astroprotein and total
protein levels in CSF of stroke patients.
enzymes, electrolytes, etc. in various neuropathological conditions,12 but there have yet been no
reports on changes in brain-specific protein levels in
CSF of stroke patients. Figure 9 demonstrates that
there was poor correlation between astroprotein and
total protein levels in CSF. The result would indicate
that the determination of astroprotein in CSF might
have a different significance than total protein.
Evidence suggests that the measurement of
astroprotein in CSF in the acute stage of stroke might
have clinical usefulness.
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Neurochemistry (Tokyo) 9: 75-78, 1970
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Disorders of Sphingolipid Metabolism. New York, Pergamon
Press, 273-278, 1966
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9. Mori T, Morimoto K, Ushio Y, Hayakawa T, Mogami H:
Radioimmunoassay of astroprotein (an astrocyte-specific
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Sekiguchi K: Radioimmunoassay of astroprotein (an astrocytespecific cerebroprotein) in cerebrospinal fluid and its clinical
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12. Engelhardt P, Avenarius HJ: Der diagnostische Wert von Enzymbestimmungen in Liquor cerebrospinalis. Med Klin 7 1 :
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Levels in stroke patients of CSF astroprotein, an astrocyte-specific cerebroprotein.
T Hayakawa, Y Ushio, T Mori, N Arita, T Yoshimine, Y Maeda, K Shimizu and A Myoga
Stroke. 1979;10:685-689
doi: 10.1161/01.STR.10.6.685
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