CEREBROSPINAL
FLUID IMMUNOGLOBULINS IN
CYSTICERCOSIS OF T H E CENTRAL NERVOUS SYSTEM
A. J.
SPINA-FRANÇA* A . LIVRAMENTO * *
L.
A . BACHESCHI***
P.
GARCIA-LOPES * * * *
Immunoglobulin G ( I G G ) is found in a higher proportion in the normal cerebrospinal fluid M
( C S F ) than
immunoglobulin A
(IGA).
Immunoglobulin
( I G M ) has been found almost only in pathological conditions.
In several
different affections of the central nervous system ( C N S ) and/or its coverings which are accompanied by an increase of gamma globulin in the C S F protein profile
— that
characterizes
the so-called chronic inflammatory profile
—
there also occurs a predominance of I G G over the other immunoglobulins. This predominance of I G G undergoes some fluctuations: many of them are proper to the disease considered, and other to its respective stage of evolu2
3
6
tion !, , , . CSF
n
.
changes proper to cysticercosis
of the C N S
5
are accompanied by
changes in the protein profile that are proper to the chronic inflammatory type also.
Data on the subject were first reported in 1 9 6 0 s
9
confirmed by other investigations > .
1 4
.
1 3
and were later
However, no studies were reported
with a view to stablishing the behavior of C S F immunoglobulins in of
cases
cysticercosis. This investigation
aims at
evaluating
the
IGG, I G A and I G M in cases of cysticercosis
MATERIAL
AND
behaviour of
C S F globulins
of the C N S .
METHODS
Cases of cysticercosis of the C N S whose confirmed diagnoses were based upon the positive result of complement fixation test for cysticercosis ( C F T ) in the CSF were studied. N o n e of the patients had undergone neurosurgical manipulation prior to this investigation. T w o series of cases are comprised in this study, the first one composed of 3 0 C S F samples, and the second of 5 . In the first series (cases 1 to 30) the positive C F T ; total protein concentration;
CSF samples were studied as t o : titer of electrophoretic profile of proteins; a g a r -
F r o m the Neurologic Investigations Center, D e p a r t m e n t of Neuro-Psychiatry (Division of N e u r o l o g y ) , São Paulo University Medical School: * Associate Professor; ** Medical Assistant; * * * Assistant Professor; * * * * Professor of Neurology, University of Londrina Medical School. This FAPESP
investigation w a s partially supported by research grant number F u n d a ç ã o rip Amnarn à P p c n n i c a rln TTctnrtn rio Q 5 n •001110
417
from
diffusion against immuneserum for I G A , I G M and IGG, first without concentration of s a m p l e and then after a 10-time previous concentration; I G A , I G M and I G G concentration. In the second series (cases 3 1 to 3 5 ) it w a s established the relative proportion of I G A , I G M and IGG, starting from the concentration values found in CSF samples concentrated around ICO t i m e s . 4
T h e titer of positive C F T w a s established according to the K o l m e r t e c h n i q u e ; results were expressed in units. Total protein concentration w a s determined through the trichloroacetic acid method according to standardization previously r e p o r t e d . The protein profile was analyzed through electrophoresis, and according to the technique previously reported ; previous concentration of C S F s a m p l e w a s carried out through dialysis against g u m arabic; cellulose acetate gel strips (Cellogel, Chemetron) were employed. Concentrations of albumin fraction and g a m m a globulin fraction were calculated considering their relative proportion in the protein profile and the total protein concentration of the C S F s a m p l e . 15
1 2
10
Agardiffusion w a s carried out according to Ouchterlony t e c h n i q u e : 2 0 m i croliters of the sample and 2 0 microliters of i m m u n e s e r u m anti I G A , I G M or IGG, produced in rabbits ( B e h r i n g w e r k e ) , were used; results are expressed with a plus ( + ) sign when precipitation line w a s evidenced, and with a minus ( - ) sign when not-evidenced. I G A , I G M and IGG concentrations were evaluated through radial immunodiffu sion technique according to M a n c i n i , and Behringwerke immunoplates were used: LC-type in the first series, using 2 0 microliters of each CSF sample, and 2 0 micro liters of three different dilutions of standard serum ( B e h r i n g w e r k e ) ; Tripartigen-type in the second series, using 5 microliters of each C S F s a m p l e concentrated around 100 times, and 5 microliters of three different dilutions of the standard serum already mentioned. W i t h base on the results obtained for the standard serum, it w a s calculated the concentration of each immunoglobulin in the samples of CSF which were tested. For the second series, the proportion of I G A , I G M and IGG was established, with base on the concentration found for each one of them. 7
For the first series, it w a s calculated the correlation (r) between: C F T titer and albumin concentration; C F T titer and g a m m a globulin concentration; C F T titer and I G G concentration; g a m m a globulin concentration and I G G concentration. Differences between correlation values were calculated for z(r) of Fisher and through Student's í test; significance w a s evaluated according to Snedocor's t a b l e .
RESULTS For the 3 0 cases of first series, results referring to titer of positive CFT, con centrations of total proteins, albumin, g a m m a globulin, I G A , I G M and I G G are presented in table 1 . Evidence of I G A , I G M and I G G through agardiffusion for non-concentrated CSF samples and for 10-time concentrated samples is presented in table 1, a l s o . T a b l e 2 shows the relative concentrations found for I G A , I G M and I G G for samples of the second series (5 cases), which were concentrated around 1 0 0 t i m e s . Analyzed correlation values are shown in table 3 .
COMMENTS
Investigations of
cysticercosis
of
on the increase of gamma globulin in the C S F in cases the
C N S already m e n t i o n e d
13
showed
that
there
is
a
positive correlation between the concentration of gamma globulin in the C S F and the respective titer of positive C F T .
Data gathered in the present in
vestigation support these findings since, for the first series of cases, it was found also a positive correlation between these data (r =
0.60; P
this
gamma
0.05). same
found were
value
globulin
and and
the titer
A s a counterproof it titer
not
its
and
significant
the (r
albumin =
0.13;
0 . 0 5 ) and this demonstrates that there is no correlation between
the
two of them, as it was expected. Presence of I G G was a constant, as can be seen in data referring to the first series of cases; in the average ( 6 . 2 m g m / 1 0 0 m l ) it proved to be superior to normal values
reported in
6
1 1
literature > .
In this
same
series
I G G represented an average of 1 6 . 1 % of total protein concentration; gamma globulin concentration represented an average of 2 0 . 1 % of the total protein concentration.
I G G concentration represented also an average of 8 7 . 9 % of
the gamma globulin concentration.
There was a significant
positive corre
lation between the representative values of I G G concentration and gamma globulin concentration (r = 0 . 6 7 ; P
FLUID IMMUNOGLOBULINS IN
CYSTICERCOSIS OF T H E CENTRAL NERVOUS SYSTEM
A. J.
SPINA-FRANÇA* A . LIVRAMENTO * *
L.
A . BACHESCHI***
P.
GARCIA-LOPES * * * *
Immunoglobulin G ( I G G ) is found in a higher proportion in the normal cerebrospinal fluid M
( C S F ) than
immunoglobulin A
(IGA).
Immunoglobulin
( I G M ) has been found almost only in pathological conditions.
In several
different affections of the central nervous system ( C N S ) and/or its coverings which are accompanied by an increase of gamma globulin in the C S F protein profile
— that
characterizes
the so-called chronic inflammatory profile
—
there also occurs a predominance of I G G over the other immunoglobulins. This predominance of I G G undergoes some fluctuations: many of them are proper to the disease considered, and other to its respective stage of evolu2
3
6
tion !, , , . CSF
n
.
changes proper to cysticercosis
of the C N S
5
are accompanied by
changes in the protein profile that are proper to the chronic inflammatory type also.
Data on the subject were first reported in 1 9 6 0 s
9
confirmed by other investigations > .
1 4
.
1 3
and were later
However, no studies were reported
with a view to stablishing the behavior of C S F immunoglobulins in of
cases
cysticercosis. This investigation
aims at
evaluating
the
IGG, I G A and I G M in cases of cysticercosis
MATERIAL
AND
behaviour of
C S F globulins
of the C N S .
METHODS
Cases of cysticercosis of the C N S whose confirmed diagnoses were based upon the positive result of complement fixation test for cysticercosis ( C F T ) in the CSF were studied. N o n e of the patients had undergone neurosurgical manipulation prior to this investigation. T w o series of cases are comprised in this study, the first one composed of 3 0 C S F samples, and the second of 5 . In the first series (cases 1 to 30) the positive C F T ; total protein concentration;
CSF samples were studied as t o : titer of electrophoretic profile of proteins; a g a r -
F r o m the Neurologic Investigations Center, D e p a r t m e n t of Neuro-Psychiatry (Division of N e u r o l o g y ) , São Paulo University Medical School: * Associate Professor; ** Medical Assistant; * * * Assistant Professor; * * * * Professor of Neurology, University of Londrina Medical School. This FAPESP
investigation w a s partially supported by research grant number F u n d a ç ã o rip Amnarn à P p c n n i c a rln TTctnrtn rio Q 5 n •001110
417
from
diffusion against immuneserum for I G A , I G M and IGG, first without concentration of s a m p l e and then after a 10-time previous concentration; I G A , I G M and I G G concentration. In the second series (cases 3 1 to 3 5 ) it w a s established the relative proportion of I G A , I G M and IGG, starting from the concentration values found in CSF samples concentrated around ICO t i m e s . 4
T h e titer of positive C F T w a s established according to the K o l m e r t e c h n i q u e ; results were expressed in units. Total protein concentration w a s determined through the trichloroacetic acid method according to standardization previously r e p o r t e d . The protein profile was analyzed through electrophoresis, and according to the technique previously reported ; previous concentration of C S F s a m p l e w a s carried out through dialysis against g u m arabic; cellulose acetate gel strips (Cellogel, Chemetron) were employed. Concentrations of albumin fraction and g a m m a globulin fraction were calculated considering their relative proportion in the protein profile and the total protein concentration of the C S F s a m p l e . 15
1 2
10
Agardiffusion w a s carried out according to Ouchterlony t e c h n i q u e : 2 0 m i croliters of the sample and 2 0 microliters of i m m u n e s e r u m anti I G A , I G M or IGG, produced in rabbits ( B e h r i n g w e r k e ) , were used; results are expressed with a plus ( + ) sign when precipitation line w a s evidenced, and with a minus ( - ) sign when not-evidenced. I G A , I G M and IGG concentrations were evaluated through radial immunodiffu sion technique according to M a n c i n i , and Behringwerke immunoplates were used: LC-type in the first series, using 2 0 microliters of each CSF sample, and 2 0 micro liters of three different dilutions of standard serum ( B e h r i n g w e r k e ) ; Tripartigen-type in the second series, using 5 microliters of each C S F s a m p l e concentrated around 100 times, and 5 microliters of three different dilutions of the standard serum already mentioned. W i t h base on the results obtained for the standard serum, it w a s calculated the concentration of each immunoglobulin in the samples of CSF which were tested. For the second series, the proportion of I G A , I G M and IGG was established, with base on the concentration found for each one of them. 7
For the first series, it w a s calculated the correlation (r) between: C F T titer and albumin concentration; C F T titer and g a m m a globulin concentration; C F T titer and I G G concentration; g a m m a globulin concentration and I G G concentration. Differences between correlation values were calculated for z(r) of Fisher and through Student's í test; significance w a s evaluated according to Snedocor's t a b l e .
RESULTS For the 3 0 cases of first series, results referring to titer of positive CFT, con centrations of total proteins, albumin, g a m m a globulin, I G A , I G M and I G G are presented in table 1 . Evidence of I G A , I G M and I G G through agardiffusion for non-concentrated CSF samples and for 10-time concentrated samples is presented in table 1, a l s o . T a b l e 2 shows the relative concentrations found for I G A , I G M and I G G for samples of the second series (5 cases), which were concentrated around 1 0 0 t i m e s . Analyzed correlation values are shown in table 3 .
COMMENTS
Investigations of
cysticercosis
of
on the increase of gamma globulin in the C S F in cases the
C N S already m e n t i o n e d
13
showed
that
there
is
a
positive correlation between the concentration of gamma globulin in the C S F and the respective titer of positive C F T .
Data gathered in the present in
vestigation support these findings since, for the first series of cases, it was found also a positive correlation between these data (r =
0.60; P
this
gamma
0.05). same
found were
value
globulin
and and
the titer
A s a counterproof it titer
not
its
and
significant
the (r
albumin =
0.13;
0 . 0 5 ) and this demonstrates that there is no correlation between
the
two of them, as it was expected. Presence of I G G was a constant, as can be seen in data referring to the first series of cases; in the average ( 6 . 2 m g m / 1 0 0 m l ) it proved to be superior to normal values
reported in
6
1 1
literature > .
In this
same
series
I G G represented an average of 1 6 . 1 % of total protein concentration; gamma globulin concentration represented an average of 2 0 . 1 % of the total protein concentration.
I G G concentration represented also an average of 8 7 . 9 % of
the gamma globulin concentration.
There was a significant
positive corre
lation between the representative values of I G G concentration and gamma globulin concentration (r = 0 . 6 7 ; P
