- Email: [email protected]
A new monoclonal antibody, GR-ICOR-3, against chick embryo myocardiac cells
327
328
F~,J.E. ,Ar~,A. ,C~,T. ,Ar~, rez,L., S s ~ , A and Gsrrido, F. Seec. ~ . l~c. ~ . Uniwe~s~ty c£ Granada. Spain.
A.E.,A1~a-
Ferr~md~z, J.E., Ar~ega,A., AIv~-ez,L.,Ar~e~m,A.E., Lama,J. ,C~rg~lez,F.J. and Gsrrido, F. Seoc. Invest. Hasic. (l~dio~asc. Faculty c~ Medicine. Uhi~'~ity o~ Grmmda..%9~in.
Faculty ~f Medicine.
~9~3 class I and II 8ntigens(DR,DP,DQ) were studied in pectineal heart muscle in order to fur~qer characterize their physiopatological and surzlcal significance in atria] muscle. The expressicn of these molecules on cells surfaces has been appr~oched to date in few studies(Dsr~ et a].,1984). A total of 57 peetinesl muscle biopsy specimens were obtained during surgery for vascu/ar diseases affecting the atria, and analyzed with the ~,~b %~5/32,G~ll,twoHLA B locus specific and a groqo of ailosz~cific NAbs obtained at the " X Internaticr~l Workshop on HLA Antigens". against class I antigens. Class II antigens were studied with GRBI,TLi22 and 57/21 to detect DR,DQ and DP. Alkaline inmundnistochemicals assays were performed in cryostatic sect/on of auricular heart muscle which was found to express HIA class I sntigems strc~gly and HLA class II antigens weakly. Our results disagree with the finding of Dar~ and cols, who recorded wesk class I antigen expression and no expression . of class II molecules in cardiac n~scle, failing to specify ~/nether atrial c~ ventricular tissue wms tested.
Despite the numerous advences in cardiac surgery in the last decade, which have provided ab~%dant methodelozical and technical information of use in heart transplants, the number of post-tran~-plant rejections is still considerable. Class I(inp]icated in allotrmnsplant rejectian) ~nd class II ~.~C antigens were studied in ventricular ffNocardium in an attempt to shed li~ht on ~lis problem in the context of earlier studies (Dsrr et al. ,1984) claindng poor expression of these membrane glycoproteins by heart muscle cells. A total of 42 biopsy speci.mns obtained during valve surgery for a variety of disceders were analyzed with the ~.~:~s W6/32(Bemstable et ai.,1978); ~ produced by Dr. L6pez Nevot.These ~zing class I antigens. ~F40 GRBI reonp/nzing a mcrgmnorphic determinant of HLA-DR antigens(produced by Dra. Cabrera). Alkaline Immm%~hc~phatsse analyses shewed strcr~ expression of HLA class I and II molecules on human n]yocardial fiber cells. The obvious presence of these molecules, particularly class I antigens,may help to elucidate lhe n~_hani~ns involved
in heart transDlent rejection.
This investigat/cns was f~ded finantiated by tl~ant Res.no.0660 and Research Ccnt~act of Biotechnolo~y Action Program (HAP) C.E.E. no.Oa67. E.
Work finant/ated by G~ant Res. no.0660 and Research Contract of Biotednnolo~y Action Program (BAP) C.E.E.. No.O467.E.
329
330
A Nmd M[I~(XONAL ANrlmJM, GR-IIIB~, GHIOK ~ M]I~A~DIAC omJS. Fe,~%mndezJ Argn~a, A . E . , ~ ] ~ , F.J., A~gnega'A" E., Mkw~s, M.A., %~lez, C., (;am-idD, F., A I ~ , L .
C h a n g i n g R e l a t i o n s h i p b e t w e e n Cell M e m b r a n e and Collagen Fibers during Development of t h e Myocardium. J.P. van Groningen and A.C.G. Wenink, Dept. of Anatomy, University of Leiden.
Seoc.
In~stig.
~ .
Craniums.
Feollty
of
Wmdicine. Univemsity of C,~--~..qpR4n.
H y p e r t r o p h i c c a r d i o m y o p a t h y is c h a r a c t e r i z e d by cellular hypertrophy, fibrosis and disorganization. In o r d e r to e l u c i d a t e which factors determine the relationship between m y o c y t e cell s u r f a c e a n d c o l l a g e n n e t w o r k we w a n t to d e s c r i b e n o r m a l d e v e l o p m e n t . In a s t e r e o l o g i c a l s t u d y o f t h e m y o c a r d i u m of 24 r a t s of d i f f e r e n t a g e s , we s t u d i e d t h e r e l a t i o n s h i p o f c o l l a g e n a n d m y o c y t e cell m e m b r a n e s . We d e t e r m i n e d t h e s u r f a c e d e n s i t y o f myocardial cell membranes. This surface density decreases rapidly until around 4 w e e k s p o s t p a r t u m a n d a f t e r t h a t more slowly u n t i l a d u l t h o o d . E a r l i e r s t e r e o l o g i c a l research showed that the amount of collagen per volume myocardiurn increases until 4 weeks post partum and remains constant a f t e r t h a t . S i n c e t h e a v a i l a b l e cell surface decreases and the amount of collagen attached to it i n c r e a s e s , the relationship b e t w e e n t h e s e two m u s t c h a n g e . S i n c e collag e n a t t a c h e s to t h e cell m e m b r a n e v i a s p e c i f ic r e c e p t o r s , t h e i n c r e a s e in t h e a m o u n t o f t h e s e c h a r a c t e r i z e s a n i m p o r t a n t p e r i o d in t h e d e v e l o p m e n t o f t h e cell m e m b r a n e .
An immunization protocol similar to that of Stahli et. al. (1980), was used. Over a total period of 24 days, four immunising injections were given to mice (Balb/c).The injectione were made up of miriced embryonic hearts (heart loop only) wieh were centrifuged in order to obtain a pellet wich was then combined with 0,2 ml. PBS. Immunizations was carried out st 7 days intervals, except for the final stimulation which took place only three days after the preceding inyection. Cells used in fusion studies were obtained from the non-Ig secreting myelema cell line Sp/15 and spleen cells from previously immunized Balb/c mice. After titulation of the MoAb, GR-ICOR-3, the supernatant was screened using IIF techniques alcng with ELISA immunoenzymatic assays. To summarize, the MoAb, GR-ICOR-3 reacted with the embryonal chick cardiac cells intraeytoplasmatically, and an intens c r o s s - r e a c t i o n was also noted with human 8trium heart musole. Action Program (BAP) C.E.E. n2 0467.E.
$98
328
F~,J.E. ,Ar~,A. ,C~,T. ,Ar~, rez,L., S s ~ , A and Gsrrido, F. Seec. ~ . l~c. ~ . Uniwe~s~ty c£ Granada. Spain.
A.E.,A1~a-
Ferr~md~z, J.E., Ar~ega,A., AIv~-ez,L.,Ar~e~m,A.E., Lama,J. ,C~rg~lez,F.J. and Gsrrido, F. Seoc. Invest. Hasic. (l~dio~asc. Faculty c~ Medicine. Uhi~'~ity o~ Grmmda..%9~in.
Faculty ~f Medicine.
~9~3 class I and II 8ntigens(DR,DP,DQ) were studied in pectineal heart muscle in order to fur~qer characterize their physiopatological and surzlcal significance in atria] muscle. The expressicn of these molecules on cells surfaces has been appr~oched to date in few studies(Dsr~ et a].,1984). A total of 57 peetinesl muscle biopsy specimens were obtained during surgery for vascu/ar diseases affecting the atria, and analyzed with the ~,~b %~5/32,G~ll,twoHLA B locus specific and a groqo of ailosz~cific NAbs obtained at the " X Internaticr~l Workshop on HLA Antigens". against class I antigens. Class II antigens were studied with GRBI,TLi22 and 57/21 to detect DR,DQ and DP. Alkaline inmundnistochemicals assays were performed in cryostatic sect/on of auricular heart muscle which was found to express HIA class I sntigems strc~gly and HLA class II antigens weakly. Our results disagree with the finding of Dar~ and cols, who recorded wesk class I antigen expression and no expression . of class II molecules in cardiac n~scle, failing to specify ~/nether atrial c~ ventricular tissue wms tested.
Despite the numerous advences in cardiac surgery in the last decade, which have provided ab~%dant methodelozical and technical information of use in heart transplants, the number of post-tran~-plant rejections is still considerable. Class I(inp]icated in allotrmnsplant rejectian) ~nd class II ~.~C antigens were studied in ventricular ffNocardium in an attempt to shed li~ht on ~lis problem in the context of earlier studies (Dsrr et al. ,1984) claindng poor expression of these membrane glycoproteins by heart muscle cells. A total of 42 biopsy speci.mns obtained during valve surgery for a variety of disceders were analyzed with the ~.~:~s W6/32(Bemstable et ai.,1978); ~ produced by Dr. L6pez Nevot.These ~zing class I antigens. ~F40 GRBI reonp/nzing a mcrgmnorphic determinant of HLA-DR antigens(produced by Dra. Cabrera). Alkaline Immm%~hc~phatsse analyses shewed strcr~ expression of HLA class I and II molecules on human n]yocardial fiber cells. The obvious presence of these molecules, particularly class I antigens,may help to elucidate lhe n~_hani~ns involved
in heart transDlent rejection.
This investigat/cns was f~ded finantiated by tl~ant Res.no.0660 and Research Ccnt~act of Biotechnolo~y Action Program (HAP) C.E.E. no.Oa67. E.
Work finant/ated by G~ant Res. no.0660 and Research Contract of Biotednnolo~y Action Program (BAP) C.E.E.. No.O467.E.
329
330
A Nmd M[I~(XONAL ANrlmJM, GR-IIIB~, GHIOK ~ M]I~A~DIAC omJS. Fe,~%mndezJ Argn~a, A . E . , ~ ] ~ , F.J., A~gnega'A" E., Mkw~s, M.A., %~lez, C., (;am-idD, F., A I ~ , L .
C h a n g i n g R e l a t i o n s h i p b e t w e e n Cell M e m b r a n e and Collagen Fibers during Development of t h e Myocardium. J.P. van Groningen and A.C.G. Wenink, Dept. of Anatomy, University of Leiden.
Seoc.
In~stig.
~ .
Craniums.
Feollty
of
Wmdicine. Univemsity of C,~--~..qpR4n.
H y p e r t r o p h i c c a r d i o m y o p a t h y is c h a r a c t e r i z e d by cellular hypertrophy, fibrosis and disorganization. In o r d e r to e l u c i d a t e which factors determine the relationship between m y o c y t e cell s u r f a c e a n d c o l l a g e n n e t w o r k we w a n t to d e s c r i b e n o r m a l d e v e l o p m e n t . In a s t e r e o l o g i c a l s t u d y o f t h e m y o c a r d i u m of 24 r a t s of d i f f e r e n t a g e s , we s t u d i e d t h e r e l a t i o n s h i p o f c o l l a g e n a n d m y o c y t e cell m e m b r a n e s . We d e t e r m i n e d t h e s u r f a c e d e n s i t y o f myocardial cell membranes. This surface density decreases rapidly until around 4 w e e k s p o s t p a r t u m a n d a f t e r t h a t more slowly u n t i l a d u l t h o o d . E a r l i e r s t e r e o l o g i c a l research showed that the amount of collagen per volume myocardiurn increases until 4 weeks post partum and remains constant a f t e r t h a t . S i n c e t h e a v a i l a b l e cell surface decreases and the amount of collagen attached to it i n c r e a s e s , the relationship b e t w e e n t h e s e two m u s t c h a n g e . S i n c e collag e n a t t a c h e s to t h e cell m e m b r a n e v i a s p e c i f ic r e c e p t o r s , t h e i n c r e a s e in t h e a m o u n t o f t h e s e c h a r a c t e r i z e s a n i m p o r t a n t p e r i o d in t h e d e v e l o p m e n t o f t h e cell m e m b r a n e .
An immunization protocol similar to that of Stahli et. al. (1980), was used. Over a total period of 24 days, four immunising injections were given to mice (Balb/c).The injectione were made up of miriced embryonic hearts (heart loop only) wieh were centrifuged in order to obtain a pellet wich was then combined with 0,2 ml. PBS. Immunizations was carried out st 7 days intervals, except for the final stimulation which took place only three days after the preceding inyection. Cells used in fusion studies were obtained from the non-Ig secreting myelema cell line Sp/15 and spleen cells from previously immunized Balb/c mice. After titulation of the MoAb, GR-ICOR-3, the supernatant was screened using IIF techniques alcng with ELISA immunoenzymatic assays. To summarize, the MoAb, GR-ICOR-3 reacted with the embryonal chick cardiac cells intraeytoplasmatically, and an intens c r o s s - r e a c t i o n was also noted with human 8trium heart musole. Action Program (BAP) C.E.E. n2 0467.E.
$98