IMMUNOLOGICAL COMMUNICATIONS, 4 ( 3 ) , 243-250 (1975)
MAST CELL SENSITIZING ANTIBODIES I N MYASTHENIA GRAVIS
*
Immunol Invest Downloaded from informahealthcare.com by UB Heidelberg on 11/16/14 For personal use only.
J . Schwartz, A. Eyquem & N. Vardinon Department o f Human Y i c r o b i o l o g y , Medical School, T e l - A v i v U n i v e r s i t y and Departmen: of Immuno-Hematology and Immuno-Pathology I n s t i t u t Pasteur, P a r i s .
,
Abstract Mast C e l l s S e n s i t i z i n g A n t i b o d i e s (WSAb) were d e t e c t e d i n p a t i e n t s w i t h Myasthenia Gravis ( M G r ) , by t h e I n d i r e c t Mast C e l l D e g r a n u l a t i o n (IMCD). I n most o f t h e examined s e r a a c o r r e l a t i o n c o u l d be e s t a b l i s h e d between MCSAb and f l u o r e s c e n t a n t i b o d i e s t o muscle and t o thymus. The YCSAb were s p e c i f i c f o r muscle e x t r a c t . They were r e s i s t a n t t o prolonged h e a t i n q and t h e i r r e a c t i v i t y i n t h e IMCD t e s t was n o t complement depending. Presence o f MCSAb i n s e r a of p a t i e n t s w i t h MGr evidences t h e a u t o - a l l e r g i c f e a t u r e s o f t h i s disease. Myasthenia G r a v i s (YGr) i s c h a r a c t e r i z e d b y homologous and heteroloqous a n t i b o d i e s , d e t e c t a b l e by:
imnunofluorescence
-
to
s k e l e t a l muscles and c y t o p l a s m i c c o n s t i t u e n t s o f m e d u l l a r y c e l l s o f t h e thymus (2,4,12,16)
,-
complement f i x a t i o n (5,12,13)
, tanned
r e d c e l l a q g l u t i n a t i o n ( 3 ) and a n t i g l o b u l i n consumption (12) w i t h muscle e x t r a c t .
,
More r e c e n t l y , a c e l l u l a r a s p e c t o f hyper-
s e n s i t i v i t y has been d e s c r i b e d i n MGr (1). S i n c e t h e d i s e a s e i s c o n s i d e r e d as auto-imnune ( 1 2 , l ) we were i n t e r e s t e d t o i n v e s t i g a t e whether a n t i b o d i e s c h a r a c t e r i s t i c
24 3 Copyright 0 1975 by Marcel Dekker, Inc. All Rights Reserved. Neither this work nor any part may be reproduced or transmitted in any form or by any means, electronic or mechanical, including photocopying, microfilming, and recording, or by any information storage and retrieval system, without permission in writing from the publisher.
244
SCHWARTZ, EYQUEM, AND VARDINON
f o r hypersensitive s t a t e s , such as Past Cell S e n s i t i z i n g Antibodies (MCSPb 9 , 1 Q , l l Y l 7 , 1 8 ) can be found in s e r a of p a t i e n t s with MGr and whether t h e r e i s any c o r r e l a t i o n between t h e MCSAb and the antibodies evidenced by immunofluorescence. For this purpose, s e r a from 29 p a t i e n t s w i t h MGr, as well Immunol Invest Downloaded from informahealthcare.com by UB Heidelberg on 11/16/14 For personal use only.
as sera from r a b b i t s immunized w i t h muscle e x t r a c t from PapioPapio monkey, have been examined f o r t h e presence of MCSAb, by the I n d i r e c t Mast Cell Degranulation test (IMCD-9) and r e s u l t s compared w i t h those obtained by imunofluorescence.
Materials and Methods The hyperimmune s e r a were prepared i n t h e "Department of Immunohaematology and Imnunopathology of the I n s t i t u t Pasteur" i n P a r i s , by inoculation of r a b b i t s , w i t h muscle e x t r a c t of Papio-Papio monkey w i t h Freund's Adjuvant ( 4 ) . The i n d i r e c t immunofluorescent tests f o r d e t e c t i o n of antibodies
a g a i n s t s t r i a t e d muscles and thymus myoid c e l l s were performed i n the same Department, using c r y o s t a t s e c t i o n s of r a t diaphragm, monkey and c a l f thymus.
These s e c t i o n s were fixed by a i r only.
P o s i t i v e r e s u l t s were obtained by d e t e c t i o n , w i t h t h e microscope Labor Lux (quartz iodine lamp and i n t e r f e r e n c e f i l t e r ) , of t h e f l u o rescence of t h e I band on muscle f i b e r and of t h e cytoplasm of t h e myoid c e l l s , w i t h the l / 5 d i l u t i o n of the s e r a . P o s i t i v e r e s u l t s were obtained i n the s e r a belonging t o the 1 s t group of p a t i e n t s w i t h severe forms of Myasthenia Gravis a t the t i t e r
of 1/80 t o 1/1280, and i n the 2nd group a t t h e d i l u t i o n s of 1/5 t o 1/160 w i t h l e s s severe forms of the d i s e a s e .
245
MYASTHENIA GRAVIS
The r e s u l t s were negative i n the t h i r d group of p a t i e n t s affected w i t h mild forms of myastbenia. The IMCD t e s t ( 9 ) i s based on the property of MCSAb t o a t t a c h themselves i n v i t r o t o normal peritoneal Mast Cells ( M C ) which by f u r t h e r exposure t o the s p e c i f i c antigen undergo de-
Immunol Invest Downloaded from informahealthcare.com by UB Heidelberg on 11/16/14 For personal use only.
granulation.
The t e s t was performed by mixing a drop of p e r i -
toneal c e l l suspension from normal guinea-pigs or r a t s , w i t h 0,05ml. of sera (on s l i d e s previously stained with neutral red), and adding onto t h i s mixture 0.05ml. of a d i l u t i o n containing 10 mg/ml. of the lyophilized muscle e x t r a c t ( 6 ) . taken a f t e r 20 minutes of incubation a t 37OC.
Readings were
Cell counts c a r r i e d
out on several f i e l d s involving 40 MC each, and the proportion of degranulated F1C versus i n t a c t MC, compared w i t h t h a t i n c o n t r o l s . Readings were repeated on several s l i d e s i n t e s t and i n c o n t r o l s . The reaction was considered p o s i t i v e when average of degranulated MC i n the t e s t was a t l e a s t double t h a t in c o n t r o l s (MC and s e r a ; MC and a n t i g e n ) .
Values of p o s i t i v e t e s t s were noted from
'I+"
to
"++++", according t o the d i f f e r e n c e between t e s t and c o n t r o l s (9511,17,18).
In order t o check the s p e c i f i c i t y of the r e a c t i o n ,
t h r e e other c o n t r o l s were performed:
a ) YC, antigen and normal
s e r a ; b) MC, s e r a from p a t i e n t s with MGr and unrelated antigens (Thyroid e x t r a c t , Bovine serum albumin); MC, s e r a from P a t i e n t s w i t h other d i s e a s e s (Hashimoto d i s e a s e , Tuberculosis) , and the
muscle e x t r a c t .
SCI-IWARTZ, EYQUEM, AM) VARDINON
246
All the studies were carried out in a double blind fashion. Repeated examinations a t different days showed the results t o be reproducible.
In few cases, attempts have been made t o adsorb
the serum on muscle before testing. Results
Immunol Invest Downloaded from informahealthcare.com by UB Heidelberg on 11/16/14 For personal use only.
The 29 sera from patients with MGr were divided into three groups in accordance wi t h thei r reacti vi ty in imnunof 1uorescence :
1 . Sera positive for muscle and for thymus cells ( M t T t ) ; 2.
Sera positive for muscle and negative for thymus cells
( M t T-);
3.
Sera neqative for b o t h muscle and thymus (!I- T - ) .
As can be seen from F i g . I , the "M+T+" group showed the
hiqhest proportion of positive reactions in the IMCD and most of the positive sera reacted i n h i g h t i t e r . In the group "MtT-", the proportion o f sera which were positive in the IMCD was lower t h a n in the f i r s t group, b u t reactivity of positive sera was relatively strong. In the "M-T-"
group, most of the sera were neqative; however,
one of the two positive sera reacted with hiqh t i t e r . The sera of rabbits imnunized with Papio-Papio muscle extract responded stronqly in the IMCD t e s t . Controls with other sera or other antigens were all negative. The few sera adsorbed on muscle before testing l o s t most of their reactivity in the IMCD t e s t . In order t o see whether
thr!
reaction was complement dependent,
samples from a l l these sera were withdrawn, heated a t 56OC for 30 minutes and reexamined by the IMCD t e s t .
No changes were observed
MYASTHENIA G R A V E
247
I .
0
t+++
0
+++
0.0
Immunol Invest Downloaded from informahealthcare.com by UB Heidelberg on 11/16/14 For personal use only.
++
0
...
0
0
+ .0
1
I
M+ T+
0 0
M+ T-
1
0 0
I
M - T-
FIGURE 1 Values o f MCSAb w i t h s e r a f r o m p a t i e n t s a f f e c t e d w i t h R G r d i v i d e d i n t o t h r e e groups i n accordance w i t h t h e i r r e a c t i v i t y i n immunofluorescence. i n any o f t h e samples.
I n a d d i t i o n , i n o r d e r t o see whether t h e
a n t i b o d i e s p r e s e n t i n these s e r a were r e a g i n s , we s u b m i t t e d them t o prolonged h e a t i n g , ( 4 hours a t 56OC).
No m o d i f i c a t i o n was ob-
served a f t e r t h i s t r e a t m e n t i n t h e behaviour o f t h e WSAb.
I t s h o u l d be n o t e d t h a t no d i f f e r e n c e was observed between t h e r e s u l t s o b t a i n e d on MC from normal guinea p i q s as compared t o those o b t a i n e d w i t h MC f r o m normal r a t s .
Furthermore, r e p e a t e d
examinations on t h e same sera always l e d t o s i m i l a r r e s u l t s . Discussion Our r e s u l t s show t h a t sera from p a t i e n t s w i t h MGr (213 out o f 29), c o n t a i n MCSAb a b l e t o a t t a c h themselves i n v i t r o on normal MC, which undergo d e g r a n u l a t i o n b y f u r t h e r exposure t o t h e muscle
extract.
The r e a c t i o n i s s p e c i f i c , as can be seen f r o m t h e n e g a t i v e
r e s u l t s obtained i n the controls.
SCWARTZ, EYQUEN, AND VARDINON
248
The anti bodies are n o t reagins , considering t h e i r unaltered behaviour a f t e r prolonged heating ( 4 hours a t 56OC); their a c t i v i t y in the IMCD t e s t i s n o t complement dependent. Identification of the antibody was n o t performed; however i t has been accepted l a t e l y ,
t h a t MC can be sensitized by non-reaginic
Immunol Invest Downloaded from informahealthcare.com by UB Heidelberg on 11/16/14 For personal use only.
antibodies identified as IgG globulins (19). A correlation could be observed between the presence and t i t e r
of these antibodies and presence of fluorescent antibodies t o muscle and thymus.
This correlation was n o t absolute, and in a few cases
a lack of parallelism was found (one serum "M+T+" add two sera which did not contain MCSAh and two sera I'M-T-" which d i d
"M+T-"
contain YCSAb).
This f a c t could be exnlained by a non-identity bet-
ween these two types of antibodies
-
the fluorescent antibodies and
the MCSAb- and i s i n accordance with the findings of Beutner and collab.
(2), and of van der Geld and collab. (14), about
heterogenity of antibodies in MGr.
Our findings concerning the presence o f MCSAb t o muscle extract i n these sera evidence a hypersensitive aspect in the MGr and constitutes an immunological approach t o the understanding
of this disease. Acknowledgments We gratefully acknowledge the technical assistance of Jeanne Hermet and Sylviane Basseterre from the Department of Immunohaematol ogy and Immunopathol ogy of the I n s t i t u t Pas teur, Paris
.
249
MYASTHENIA GRAVIS
References 1.
Alpert, L.J., Rule, A . , Norio, M . , Kott, E . , Kornfeld, P . and Osserman, K.E. Am.J.Cl.patho1. 58: 647, 1972.
2.
Beutner, E.H., Witebsky, E . , Ricken, D. and Adler, R.H. J.A.M.A. E 4 6 , 1962.
Immunol Invest Downloaded from informahealthcare.com by UB Heidelberg on 11/16/14 For personal use only.
3. 4.
Djanian, A.Y.,
63: 60, 1964.
Beutner, E.H. and Witebsky, E.
J.Lab.Clin.Med.
Eyquem, A. and DUC, H.T. 2e Congrgs National de Microbiologie Medicale. Bucarest, Sept. 1971.
5. Eyquem, A. and Duc. H.T. SOC. Fransaise d'Immunologie. 13 AoOt 1972. Annales de 1 ' I n s t . Pasteur 123:115, 1972. 6.
Eyquem, A., DUC, H.T., Hermet, J. e t B a s s e t e r r e , S. SOC. Frangaise d ' Immunologie, 5-6 Avril 1973. Annales d'Immuno123: 125, 1972. logie -
7.
Grob, D. and Namba, T.
8.
Lang, R., Shulman, S., Beutner, E. and Witebsky, E. York Acad. o f Sciences. 135:601 , 1966.
9.
Schwartz, J . , Klopstock, A . , Sickert-Duvdevani, P. and Honig, S. 1nt.Arch.Allergy 26: 333, 1965.
J.Clin.Invest.
42: 940,
Experientia
1963. Ann.New-
10.
Schwartz, J. and Vardinon, N.
11.
Schwartz, J . , Vardinon, N . and Moscovici Med.Sci. 22, 1971.
12.
S t r a u s s , A. , S e e g a l , B . C . , H S U , K.C. , Burkholder, P.M., Nastuk, W.L. and Osserman, K.E. Proc.Soc.Exp.Bio1. (NY) 105: 184, 1960.
13.
S t r a u s s , A.,
14.
van d e r Geld, H . , Feltkamp, T.E.W., Van Loghem, J . J . , Oosterhius, H.J.G.H. and Biemond, A . Vox Sang & 373, 1963.
15.
van d e r Geld, H . and Oosterhuis, H . J . G . H .
16.
van d e r Geld, H . , Feltkamp. T.E.W. and O o s t e r h u i s H.J.G.H. Proc.Soc.Exper.Bio1. (NY) 115:782, 1964.
17.
Vardinon, N. and Schwartz, J.
Deitch, A. and HSU, K.
702, 1966.
, 0.
Israel J. o f
Fed.Proc. 20: 38, 1961.
Experientia
8: 196, 1963. Vox Sang, -
27: 471 , 1971.
Immunol Invest Downloaded from informahealthcare.com by UB Heidelberg on 11/16/14 For personal use only.
250
SCHMARTZ, EXQUEM, AND VARDINON
18.
Vardinon, N. and Schwartz, J . , and Ben-Efraim, S. Med.Sci. 14, 1971.
1 s r . J . of
19.
Proceedings of the Second I n t e r n a t i o n a l Congress of Immunology 21-27 J u l y 1974,Workshop 79, Brighton.
MAST CELL SENSITIZING ANTIBODIES I N MYASTHENIA GRAVIS
*
Immunol Invest Downloaded from informahealthcare.com by UB Heidelberg on 11/16/14 For personal use only.
J . Schwartz, A. Eyquem & N. Vardinon Department o f Human Y i c r o b i o l o g y , Medical School, T e l - A v i v U n i v e r s i t y and Departmen: of Immuno-Hematology and Immuno-Pathology I n s t i t u t Pasteur, P a r i s .
,
Abstract Mast C e l l s S e n s i t i z i n g A n t i b o d i e s (WSAb) were d e t e c t e d i n p a t i e n t s w i t h Myasthenia Gravis ( M G r ) , by t h e I n d i r e c t Mast C e l l D e g r a n u l a t i o n (IMCD). I n most o f t h e examined s e r a a c o r r e l a t i o n c o u l d be e s t a b l i s h e d between MCSAb and f l u o r e s c e n t a n t i b o d i e s t o muscle and t o thymus. The YCSAb were s p e c i f i c f o r muscle e x t r a c t . They were r e s i s t a n t t o prolonged h e a t i n q and t h e i r r e a c t i v i t y i n t h e IMCD t e s t was n o t complement depending. Presence o f MCSAb i n s e r a of p a t i e n t s w i t h MGr evidences t h e a u t o - a l l e r g i c f e a t u r e s o f t h i s disease. Myasthenia G r a v i s (YGr) i s c h a r a c t e r i z e d b y homologous and heteroloqous a n t i b o d i e s , d e t e c t a b l e by:
imnunofluorescence
-
to
s k e l e t a l muscles and c y t o p l a s m i c c o n s t i t u e n t s o f m e d u l l a r y c e l l s o f t h e thymus (2,4,12,16)
,-
complement f i x a t i o n (5,12,13)
, tanned
r e d c e l l a q g l u t i n a t i o n ( 3 ) and a n t i g l o b u l i n consumption (12) w i t h muscle e x t r a c t .
,
More r e c e n t l y , a c e l l u l a r a s p e c t o f hyper-
s e n s i t i v i t y has been d e s c r i b e d i n MGr (1). S i n c e t h e d i s e a s e i s c o n s i d e r e d as auto-imnune ( 1 2 , l ) we were i n t e r e s t e d t o i n v e s t i g a t e whether a n t i b o d i e s c h a r a c t e r i s t i c
24 3 Copyright 0 1975 by Marcel Dekker, Inc. All Rights Reserved. Neither this work nor any part may be reproduced or transmitted in any form or by any means, electronic or mechanical, including photocopying, microfilming, and recording, or by any information storage and retrieval system, without permission in writing from the publisher.
244
SCHWARTZ, EYQUEM, AND VARDINON
f o r hypersensitive s t a t e s , such as Past Cell S e n s i t i z i n g Antibodies (MCSPb 9 , 1 Q , l l Y l 7 , 1 8 ) can be found in s e r a of p a t i e n t s with MGr and whether t h e r e i s any c o r r e l a t i o n between t h e MCSAb and the antibodies evidenced by immunofluorescence. For this purpose, s e r a from 29 p a t i e n t s w i t h MGr, as well Immunol Invest Downloaded from informahealthcare.com by UB Heidelberg on 11/16/14 For personal use only.
as sera from r a b b i t s immunized w i t h muscle e x t r a c t from PapioPapio monkey, have been examined f o r t h e presence of MCSAb, by the I n d i r e c t Mast Cell Degranulation test (IMCD-9) and r e s u l t s compared w i t h those obtained by imunofluorescence.
Materials and Methods The hyperimmune s e r a were prepared i n t h e "Department of Immunohaematology and Imnunopathology of the I n s t i t u t Pasteur" i n P a r i s , by inoculation of r a b b i t s , w i t h muscle e x t r a c t of Papio-Papio monkey w i t h Freund's Adjuvant ( 4 ) . The i n d i r e c t immunofluorescent tests f o r d e t e c t i o n of antibodies
a g a i n s t s t r i a t e d muscles and thymus myoid c e l l s were performed i n the same Department, using c r y o s t a t s e c t i o n s of r a t diaphragm, monkey and c a l f thymus.
These s e c t i o n s were fixed by a i r only.
P o s i t i v e r e s u l t s were obtained by d e t e c t i o n , w i t h t h e microscope Labor Lux (quartz iodine lamp and i n t e r f e r e n c e f i l t e r ) , of t h e f l u o rescence of t h e I band on muscle f i b e r and of t h e cytoplasm of t h e myoid c e l l s , w i t h the l / 5 d i l u t i o n of the s e r a . P o s i t i v e r e s u l t s were obtained i n the s e r a belonging t o the 1 s t group of p a t i e n t s w i t h severe forms of Myasthenia Gravis a t the t i t e r
of 1/80 t o 1/1280, and i n the 2nd group a t t h e d i l u t i o n s of 1/5 t o 1/160 w i t h l e s s severe forms of the d i s e a s e .
245
MYASTHENIA GRAVIS
The r e s u l t s were negative i n the t h i r d group of p a t i e n t s affected w i t h mild forms of myastbenia. The IMCD t e s t ( 9 ) i s based on the property of MCSAb t o a t t a c h themselves i n v i t r o t o normal peritoneal Mast Cells ( M C ) which by f u r t h e r exposure t o the s p e c i f i c antigen undergo de-
Immunol Invest Downloaded from informahealthcare.com by UB Heidelberg on 11/16/14 For personal use only.
granulation.
The t e s t was performed by mixing a drop of p e r i -
toneal c e l l suspension from normal guinea-pigs or r a t s , w i t h 0,05ml. of sera (on s l i d e s previously stained with neutral red), and adding onto t h i s mixture 0.05ml. of a d i l u t i o n containing 10 mg/ml. of the lyophilized muscle e x t r a c t ( 6 ) . taken a f t e r 20 minutes of incubation a t 37OC.
Readings were
Cell counts c a r r i e d
out on several f i e l d s involving 40 MC each, and the proportion of degranulated F1C versus i n t a c t MC, compared w i t h t h a t i n c o n t r o l s . Readings were repeated on several s l i d e s i n t e s t and i n c o n t r o l s . The reaction was considered p o s i t i v e when average of degranulated MC i n the t e s t was a t l e a s t double t h a t in c o n t r o l s (MC and s e r a ; MC and a n t i g e n ) .
Values of p o s i t i v e t e s t s were noted from
'I+"
to
"++++", according t o the d i f f e r e n c e between t e s t and c o n t r o l s (9511,17,18).
In order t o check the s p e c i f i c i t y of the r e a c t i o n ,
t h r e e other c o n t r o l s were performed:
a ) YC, antigen and normal
s e r a ; b) MC, s e r a from p a t i e n t s with MGr and unrelated antigens (Thyroid e x t r a c t , Bovine serum albumin); MC, s e r a from P a t i e n t s w i t h other d i s e a s e s (Hashimoto d i s e a s e , Tuberculosis) , and the
muscle e x t r a c t .
SCI-IWARTZ, EYQUEM, AM) VARDINON
246
All the studies were carried out in a double blind fashion. Repeated examinations a t different days showed the results t o be reproducible.
In few cases, attempts have been made t o adsorb
the serum on muscle before testing. Results
Immunol Invest Downloaded from informahealthcare.com by UB Heidelberg on 11/16/14 For personal use only.
The 29 sera from patients with MGr were divided into three groups in accordance wi t h thei r reacti vi ty in imnunof 1uorescence :
1 . Sera positive for muscle and for thymus cells ( M t T t ) ; 2.
Sera positive for muscle and negative for thymus cells
( M t T-);
3.
Sera neqative for b o t h muscle and thymus (!I- T - ) .
As can be seen from F i g . I , the "M+T+" group showed the
hiqhest proportion of positive reactions in the IMCD and most of the positive sera reacted i n h i g h t i t e r . In the group "MtT-", the proportion o f sera which were positive in the IMCD was lower t h a n in the f i r s t group, b u t reactivity of positive sera was relatively strong. In the "M-T-"
group, most of the sera were neqative; however,
one of the two positive sera reacted with hiqh t i t e r . The sera of rabbits imnunized with Papio-Papio muscle extract responded stronqly in the IMCD t e s t . Controls with other sera or other antigens were all negative. The few sera adsorbed on muscle before testing l o s t most of their reactivity in the IMCD t e s t . In order t o see whether
thr!
reaction was complement dependent,
samples from a l l these sera were withdrawn, heated a t 56OC for 30 minutes and reexamined by the IMCD t e s t .
No changes were observed
MYASTHENIA G R A V E
247
I .
0
t+++
0
+++
0.0
Immunol Invest Downloaded from informahealthcare.com by UB Heidelberg on 11/16/14 For personal use only.
++
0
...
0
0
+ .0
1
I
M+ T+
0 0
M+ T-
1
0 0
I
M - T-
FIGURE 1 Values o f MCSAb w i t h s e r a f r o m p a t i e n t s a f f e c t e d w i t h R G r d i v i d e d i n t o t h r e e groups i n accordance w i t h t h e i r r e a c t i v i t y i n immunofluorescence. i n any o f t h e samples.
I n a d d i t i o n , i n o r d e r t o see whether t h e
a n t i b o d i e s p r e s e n t i n these s e r a were r e a g i n s , we s u b m i t t e d them t o prolonged h e a t i n g , ( 4 hours a t 56OC).
No m o d i f i c a t i o n was ob-
served a f t e r t h i s t r e a t m e n t i n t h e behaviour o f t h e WSAb.
I t s h o u l d be n o t e d t h a t no d i f f e r e n c e was observed between t h e r e s u l t s o b t a i n e d on MC from normal guinea p i q s as compared t o those o b t a i n e d w i t h MC f r o m normal r a t s .
Furthermore, r e p e a t e d
examinations on t h e same sera always l e d t o s i m i l a r r e s u l t s . Discussion Our r e s u l t s show t h a t sera from p a t i e n t s w i t h MGr (213 out o f 29), c o n t a i n MCSAb a b l e t o a t t a c h themselves i n v i t r o on normal MC, which undergo d e g r a n u l a t i o n b y f u r t h e r exposure t o t h e muscle
extract.
The r e a c t i o n i s s p e c i f i c , as can be seen f r o m t h e n e g a t i v e
r e s u l t s obtained i n the controls.
SCWARTZ, EYQUEN, AND VARDINON
248
The anti bodies are n o t reagins , considering t h e i r unaltered behaviour a f t e r prolonged heating ( 4 hours a t 56OC); their a c t i v i t y in the IMCD t e s t i s n o t complement dependent. Identification of the antibody was n o t performed; however i t has been accepted l a t e l y ,
t h a t MC can be sensitized by non-reaginic
Immunol Invest Downloaded from informahealthcare.com by UB Heidelberg on 11/16/14 For personal use only.
antibodies identified as IgG globulins (19). A correlation could be observed between the presence and t i t e r
of these antibodies and presence of fluorescent antibodies t o muscle and thymus.
This correlation was n o t absolute, and in a few cases
a lack of parallelism was found (one serum "M+T+" add two sera which did not contain MCSAh and two sera I'M-T-" which d i d
"M+T-"
contain YCSAb).
This f a c t could be exnlained by a non-identity bet-
ween these two types of antibodies
-
the fluorescent antibodies and
the MCSAb- and i s i n accordance with the findings of Beutner and collab.
(2), and of van der Geld and collab. (14), about
heterogenity of antibodies in MGr.
Our findings concerning the presence o f MCSAb t o muscle extract i n these sera evidence a hypersensitive aspect in the MGr and constitutes an immunological approach t o the understanding
of this disease. Acknowledgments We gratefully acknowledge the technical assistance of Jeanne Hermet and Sylviane Basseterre from the Department of Immunohaematol ogy and Immunopathol ogy of the I n s t i t u t Pas teur, Paris
.
249
MYASTHENIA GRAVIS
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