CEREBROSPINAL FLUID IMMUNOGLOBULINS IN
CYSTICERCOSIS OF T H E C E N T R A L NERVOUS S Y S T E M
A . S P I N A - F R A N Ç A * J . A . L I V R A M E N T O * * L . A . B A C H E S C H I * * * P . G A R C I A - L O P E S * * * *
Immunoglobulin G ( I G G ) is found in a higher proportion in the normal cerebrospinal fluid ( C S F ) than immunoglobulin A ( I G A ) . Immunoglobulin M ( I G M ) has been found almost only in pathological conditions. In several different affections of the central nervous system ( C N S ) a n d / o r its coverings which are accompanied by an increase of g a m m a 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 evolu-tion !, 2
, 3
, 6
.n
.
C S F changes proper to cysticercosis of the C N S5
are accompanied by changes in the protein profile that are proper to the chronic inflammatory type also. D a t a on the subject were first reported in 1 9 6 01 3
and were later confirmed by other investigations s
> 9
.1 4
. However, no studies were reported with a view to stablishing the behavior of C S F immunoglobulins in cases of cysticercosis.
This investigation aims at evaluating the behaviour of C S F globulins IGG, I G A and I G M in cases of cysticercosis of the C N S .
M A T E R I A L A N D M E T H O D S
Cases of cysticercosis of the C N S w h o s e confirmed diagnoses were based upon the positive result of c o m p l e m e n t fixation test for cysticercosis ( C F T ) in the C S F were studied. N o n e of the patients had undergone neurosurgical m a n i p u l a t i o n 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 3 0 ) the C S F samples were studied as t o : titer of positive C F T ; total protein concentration; electrophoretic profile of proteins; a g a r
-F r o m the N e u r o l o g i c I n v e s t i g a t i o n s Center, D e p a r t m e n t of N e u r o - P s y c h i a t r y (Division of N e u r o l o g y ) , São P a u l o University M e d i c a l School: * A s s o c i a t e Professor; ** M e d i c a l A s s i s t a n t ; * * * A s s i s t a n t Professor; * * * * Professor of N e u r o l o g y , Univer-sity of Londrina M e d i c a l School.
diffusion a g a i n s t i m m u n e s e r u m for I G A , I G M and I G G , first without concentration of s a m p l e and then after a 1 0 - t i m e previous concentration; I G A , I G M and I G G concentration. I n 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 I G G , starting from the concentration v a l u e s found in CSF samples concentrated around ICO t i m e s .
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 e4
; results were expressed in units. T o t a l protein concentration w a s determined through the trichloroacetic acid method according to standardization previously r e p o r t e d1 5
. The protein profile w a s analyzed t h r o u g h electrophoresis, and according to the technique previously reported 1 2
; previous concentration of C S F s a m p l e w a s carried out t h r o u g h dialysis a g a i n s t g u m arabic; cellulose acetate gel strips (Cellogel, C h e -m e t r o n ) were e -m p l o y e d . Concentrations of a l b u -m i n fraction and g a -m -m a globulin fraction were c a l c u l a t e d considering their relative proportion in the protein profile and the total protein concentration of the C S F s a m p l e .
A g a r d i f f u s i o n w a s carried out according to Ouchterlony t e c h n i q u e1 0
: 2 0 m i -croliters of the s a m p l e and 2 0 mi-croliters of i m m u n e s e r u m anti I G A , I G M or I G G , produced in rabbits ( B e h r i n g w e r k e ) , were used; results are expressed with a plus ( + ) sign w h e n precipitation line w a s evidenced, and with a minus ( - ) sign when not-evidenced.
I G A , I G M and I G G concentrations were e v a l u a t e d t h r o u g h radial i m m u n o d i f f u -sion technique according to M a n c i n i7
, and B e h r i n g w e r k e i m m u n o p l a t e s were used: L C - t y p e in the first series, using 2 0 microliters of each C S F 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 1 0 0 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 i m m u n o g l o b u l i n in the s a m p l e s of CSF which w e r e tested. For the second series, the proportion of I G A , I G M and IGG w a s established, with base on the concentration found for each one of them.
For the first series, it w a s c a l c u l a t e d the correlation ( r ) b e t w e e n : C F T titer and a l b u m i n 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 v a l u e s were c a l c u l a t e d for z(r) of Fisher and through Student's í test; significance w a s e v a l u a t e d according to Snedocor's t a b l e .
R E S U L T S
For t h e 3 0 cases of first series, results referring to titer of positive C F T , con-centrations of total proteins, a l b u m i n , g a m m a globulin, I G A , I G M and I G G are presented in t a b l e 1 . Evidence of I G A , I G M and I G G t h r o u g h agardiffusion for non-concentrated C S F s a m p l e s and for 1 0 - t i m e 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 s a m p l e s of the second series ( 5 c a s e s ) , which were concentrated around 1 0 0 t i m e s .
A n a l y z e d correlation values are shown in table 3 .
C O M M E N T S
Investigations on the increase of g a m m a globulin in the C S F in cases of cysticercosis of the C N S already m e n t i o n e d1 3
A positive correlation between C F T titer and I G G concentration was also clear (r = 0 . 4 0 ; P < 0 . 0 5 ) . T h e difference between its value and the value found for correlation of concentration of g a m m a globulin and titer of C F T was not significative (t = 0 . 9 3 ; P > 0 . 0 5 ) . A s a counterproof it was calculated the correlation between this same titer and the albumin fraction concentration: the values found were not significant (r = 0 . 1 3 ; P > 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 literature 6
>1 1
I G G represented an average of 1 6 . 1 % of total protein concentration; g a m m a 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 g a m m a 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 < 0 . 0 1 ) .
I G A and I G M were present, in the first series of samples of the cases analyzed, as follows: in none of the cases when agardiffusion of non-concen-trated C S F was considered; I G A in one sample and I G M in three samples, when radial immunodiffusion was considered. However, when 10-time con-centrated C S F samples were used — and it was possible to do that in 16 of the cases — I G A was found to be present in 8 cases, and I G M in 2.
Through analysis of this latter series it can be verified that, with the exception of one case, presence of the three immunoglobulins studied can be demonstrated; a higher proportion of I G G ( 8 0 . 4 , average) is confirmed in relation to I G A and I G M ( 1 1 . 9 % and 7 . 7 % average, respectively).
These data prove that for an appropriate study of events characteristic of I G A and I G M , in inflammatory processes which produce immunobiolo-gical modifications in the C S F similar to those found in cysticercosis of the C N S , it is necessary to use properly concentrated samples or more precise methods than those currently available — and hereby analyzed.
D a t a obtained as to I G G show it to be the main immunoglobulin occuring in the C S F in cases of cysticercosis of the C N S . In such cases, it is part of the protein profile in a proportion higher than the usually referred in normal C S F . In this series, this proportion was of 16%, representing about 88% of the g a m m a globulin concentration. There is an unmistakable corre-lation between g a m m a globulin concentration and I G G concentration — and also between the latter and the C F T positivity titer.
S U M M A R Y
Investigation on the behavior of immunoglobulins I G G , I G A and I G M in the C S F in cases of cysticercosis of the C N S , based on data pertaining to two different series of cases. T h e first series comprises 30 samples of CSF, and the second one, 5 samples. I t was demonstrated that I G G is the one representing the largest contingent. I G G concentration keeps in pro-portion with the g a m m a globulin concentration, of which it represented an 88% average in the cases studied. Participation of I G G in the protein pro-file of the C S F is greater than the usually referred; the results for the material analyzed showed 16%. I t was verified a proporcionality also between I G G concentration and the titer of positive complement fixation test for cysticercosis; there is a positive correlation, whose numerical expression was found to be significant, in the samσles studied.
R E S U M O
Imunoglobulinas do líquido cefálorraqueano na cistieercose
do sistema nervoso central
repre-sentou 88% em mιdia. A participação de I G G no perfil proteico do L C R ι maior do que o habitualmente referido, sendo de 1 6 % para o material analisado. F o i verificado t a m b ι m haver proporcionalidade entre a concentra-ção de I G G e o título da positividade da reaconcentra-ção de fixaconcentra-ção do complemento para cisticercose: hα correlação positiva, cuja expressão numιrica foi sig-nificativa para as amostras estudadas.
R E F E R E N C E S
1 . A N S A R I , K . A . ; W E L L S , K . & V A T A S S E R R Y , G. T. — Q u a n t i t a t i v e esti-m a t i o n of cerebrospinal fluid globulins in esti-multiple sclerosis. A coesti-mparison of electrophoretic and i m m u n o l o g i c m e t h o d s . N e u r o l o g y (Minneapolis) 2 5 : 6 8 8 , 1 9 7 5 .
2 . F I S C H E R - W I L L I A M S , M . & R O B E R T S , R . — Cerebrospinal fluid proteins and serum i m m u n o g l o b u l i n s . Occurrence in multiple sclerosis and other neurologic diseases: c o m p a r a t i v e m e a s u r e m e n t of g a m m a - g l o b u l i n and the I G G class. A r c h . N e u r o l . ( C h i c a g o ) 2 5 : 5 2 6 , 1 9 7 1 .
3 . G L A S N E R , H . — Barrier i m p a i r m e n t and i m m u n e reaction in the cerebrospinal f l u i d . E u r . N e u r o l . 1 3 : 3 0 4 , 1 9 7 5 .
4 . K O L M E R , J . A . ; S P A U L D I N G , E . H . & R O B I N S O N , H . W . — A p p r o v e d L a b o r a t o r y T e c h n i c . A p p l e t o n - C e n t u r y - C r o f t s Inc., N e w Y o r k , 1 9 5 1 , pg. 797. 5 . L A N G E , O . — Síndrome liquσrica da cisticercose e n c ι f a l o - m e n í n g e a . R e v .
Neurol. Psiquiat. de São P a u l o 6 : 3 5 , 1 9 4 0 .
6 . L I N K , H . & M ٢ L L E R , R . — I m m u n o g l o b u l i n s in m u l t i p l e sclerosis and in-fections of the nervous s y s t e m . A r c h . N e u r o l . ( C h i c a g o ) 2 5 : 3 2 6 , 1 9 7 1 . 7 . M A N C I N I , G . ; V A E R M A N , J . P . ; C A R B O N A R A , A . O . & H E R E M A N S , J . F.
— A single radial diffusion m e t h o d for the i m m u n o l o g i c a l quantitation of proteins. In Peters, H . (Ed.) — Protides in Biological F l u i d s . Elsevier Publ. Co., A m s t e r d a m , 1963, p g . 3 7 0 .
8 . O B E R H A U S E R , E . & W E I N S T E I N , V . — Estudio c o m p a r a t i v o de líquido cιfalo raquídeo y s a n g u e en l٥es y cisticercosis. N e u r o c i r u r g i a 2 7 : 6 1 , 1969. 9 . O R D A Z , P. — Diagnσstico de la neurocisticercosis por el e x a m e n del líquido cefalorraquídeo. T e s i s . F a c u l d a d de Medicina, Univ. Central de Venezuela. Caracas, 1 9 7 2 .
1 0 . O U C H T E R L O N Y , O. — In vitro method for testing the toxin producing ca-pacity of diphteria b a c t e r i a . A c t a p a t h , m i c r o b . s c a n d . 2 5 : 1 8 6 , 1 9 4 8 . 1 1 . S A V O R Y , J . & H E I N T G E S , M . G . — Cerebrospinal fluid levels of I G G , I G A
and I G M in neurologic diseases. N e u r o l o g y (Minneapolis) 2 3 : 9 5 3 , 1 9 7 4 . 1 2 . S P I N A - F R A N Η A , A . — Eletroforese das proteínas do líquido cefalorraqueano:
Tιcnica. A r q . Neuro-Psiquiat. (São P a u l o ) 1 6 : 2 3 6 , 1 9 5 8 .
1 3 . S P I N A - F R A N Η A , A . — V a l o r do e x a m e eletroforιtico d a s proteínas do liquido cefalorraqueano n a cisticercose do sistema nervoso c e n t r a l . A r q . Neuro-Psi-quiat. (São P a u l o ) 1 8 : 3 0 1 , 1 9 6 0 .
1 4 . S P I N A - F R A N Η A , A . — I m u n o b i o l o g i a da cisticercose: a v a l i a ç ã o dos conceitos a t u a i s . A r q . Neuro-Psiquiat. ( S ã o P a u l o ) 2 7 : 1 2 5 , 1 9 6 9 .
1 5 . S P I N A - F R A N Η A , A . & A M A R , I. —• V a l o r e s normais da concentração proteica do líquido cefalorraquidiano cisternal. A r q . Neuro-Psiquiat. ( S ã o P a u l o ) 1 5 : 27, 1 9 5 7 .
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