- Email: [email protected]
The highly significant association of non-HLA, anti-epithelial cell cytotoxic antibodies with renal transplant failure
10
Abstracts
TBEHI61ILY SIGNIFICANT ASSOCIATION OF NON-KLA, ANTI-EPITHELIAL CELL CYTOTOXIC ANTIBODIES WITH RENAL TBANSPIANT FAILURE. S Marlin, PE Brenchley, PA Dyer, RJ Postlethwai%e, RWG Johnson, NWRegional T i s s u e T y p i n g & Immunology L a b s , R o y a l C h i l d r e n ' s H o s p i t a l , R e n a l Transplant Unit, Manchester, England~ [ n o r d e r t o a s s e s s t h e r o l e o f non-}HA a n t i b o d i e s in renal transplant rejection, we d e v e l o p e d a m i c r o c y t o t e x i c i t y assay whereby the adherent epithelial c e l l l i n e A5~9 was i n c u b a t e d o v e r n i g h t i n T e r a s a k i trays and then used as the target in a eytotoxicity assay for the screening of patient sera. Percentage cell killing, indicating the presence of anti-epithelial cell antibodies (AEC), was a s s e s s e d a f t e r staining with ethidium bromide and acridine orange. 77 c h i l d r e n w e r e r e t r o s p e c t i v e l y s c r e e n e d f o r t h e p r e s e n c e o f AEC. Of ~5 w i t h f u n c t i o n i n g t r a n s p l a n t s , 56 were AEC n e g a t i v e : 7 were AEC positive and also had autoreactive lymphocytotoxic antibodies (AEC/ auto). Of t h e 52 f a i l u r e s , t e s t e d a t t h e t i m e o f n e p h r e c t o m y , 22 were AEC n e g a t i v e : 10 were AEC p o s i t i v e ; in 5 cases these were AEC/auto. 50 a d u l t s were a l s o s t u d i e d : 6 h a d AEC o f w h i c h 2 w e r e A E C / a u t o . In the children, e x c l u d i n g t h e A E C / a u t o s , 7 o f t h e 52 f a i l u r e s as c o m p a r e d w i t h n o n e o f t h e ~5 s u c c e s s f u l transplants p r o d u c e d AEC (p=0.OO17). The t r a n s p l a n t s f a i l e d i n a l l 11 p a t i e n t s w i t h AEC w h e r e a s h E C / a u t o s were f o u n d i n 5 f a i l u r e s and 7 successes (p=O.O07). Pret r e a t m e n t o f t h e s e r a w i t h 0.01M DTT showed a l l AEC t o be IgM antibodies: failure to block the reactions with a monomorphic anti}HA c l a s s I a n t i b o d y showed t h e m n o t t o be d i r e c f i e d a g a i n s t HIA c l a s s I antigens. I n c o n c l u s i o n , AEC were n e v e r a s s o c i a t e d w i t h s u c c e s s f u l transplantation b u t were p r o d u c e d by 21~ o f c h i l d r e n a n d 8~ o f a d u l t s whose t r a n s p l a n t s failed.
STUDY OF SERUM REACTION PATrERNS OF PATIENTS WHO REJECTED KIDNEY TRANSPLANTS ACCORDING TO THE AMINO ACID SEQUENCES THAT MISMATCH. BD Clark, MS Park, PI Terasaki, Department of Surgery, University of California at Los Angeles, Los Angeles, California
When screening sera of highly sensitized patients, although the conventional serum analysis programs can identify included specificities to some degree, it is often difficult to be assured of the specificities. By development of computer programs that have the capability of storing the amino acid sequences for each of the panel cells, it has been possible to identit~¢ many high frequency sera 'as being directed to a "logical" epitope, represented by a single amino acid. We examined sera from 20 patients who had rejected their transplants. In one example, the kidney donor ~as A1 A24 B7 B62 and the recipient was A3 B7 B62. Thrce months after rejection, the antibody present in the recipient was anti A1,2,24,25,26,31,32,and 33. The program identified this antibody as simply being one directed against histidinc at positon 70. In this way, many "multispecific" antisera can be calssified more simply classified according to the amino acid 'epitope' against which they are directed. Moreover, the epitopes which result after rejection of a transplant can be postulated as the important immunogenic epitopcs against which the recipient reacted. To date, because of the multispecificity, sera from patients who had rejected have been difficult to analyze. Many more sera from patients who had rejected grafts are being investigated, and the program is being extended to analyzc the Class II specificities.
Abstracts
TBEHI61ILY SIGNIFICANT ASSOCIATION OF NON-KLA, ANTI-EPITHELIAL CELL CYTOTOXIC ANTIBODIES WITH RENAL TBANSPIANT FAILURE. S Marlin, PE Brenchley, PA Dyer, RJ Postlethwai%e, RWG Johnson, NWRegional T i s s u e T y p i n g & Immunology L a b s , R o y a l C h i l d r e n ' s H o s p i t a l , R e n a l Transplant Unit, Manchester, England~ [ n o r d e r t o a s s e s s t h e r o l e o f non-}HA a n t i b o d i e s in renal transplant rejection, we d e v e l o p e d a m i c r o c y t o t e x i c i t y assay whereby the adherent epithelial c e l l l i n e A5~9 was i n c u b a t e d o v e r n i g h t i n T e r a s a k i trays and then used as the target in a eytotoxicity assay for the screening of patient sera. Percentage cell killing, indicating the presence of anti-epithelial cell antibodies (AEC), was a s s e s s e d a f t e r staining with ethidium bromide and acridine orange. 77 c h i l d r e n w e r e r e t r o s p e c t i v e l y s c r e e n e d f o r t h e p r e s e n c e o f AEC. Of ~5 w i t h f u n c t i o n i n g t r a n s p l a n t s , 56 were AEC n e g a t i v e : 7 were AEC positive and also had autoreactive lymphocytotoxic antibodies (AEC/ auto). Of t h e 52 f a i l u r e s , t e s t e d a t t h e t i m e o f n e p h r e c t o m y , 22 were AEC n e g a t i v e : 10 were AEC p o s i t i v e ; in 5 cases these were AEC/auto. 50 a d u l t s were a l s o s t u d i e d : 6 h a d AEC o f w h i c h 2 w e r e A E C / a u t o . In the children, e x c l u d i n g t h e A E C / a u t o s , 7 o f t h e 52 f a i l u r e s as c o m p a r e d w i t h n o n e o f t h e ~5 s u c c e s s f u l transplants p r o d u c e d AEC (p=0.OO17). The t r a n s p l a n t s f a i l e d i n a l l 11 p a t i e n t s w i t h AEC w h e r e a s h E C / a u t o s were f o u n d i n 5 f a i l u r e s and 7 successes (p=O.O07). Pret r e a t m e n t o f t h e s e r a w i t h 0.01M DTT showed a l l AEC t o be IgM antibodies: failure to block the reactions with a monomorphic anti}HA c l a s s I a n t i b o d y showed t h e m n o t t o be d i r e c f i e d a g a i n s t HIA c l a s s I antigens. I n c o n c l u s i o n , AEC were n e v e r a s s o c i a t e d w i t h s u c c e s s f u l transplantation b u t were p r o d u c e d by 21~ o f c h i l d r e n a n d 8~ o f a d u l t s whose t r a n s p l a n t s failed.
STUDY OF SERUM REACTION PATrERNS OF PATIENTS WHO REJECTED KIDNEY TRANSPLANTS ACCORDING TO THE AMINO ACID SEQUENCES THAT MISMATCH. BD Clark, MS Park, PI Terasaki, Department of Surgery, University of California at Los Angeles, Los Angeles, California
When screening sera of highly sensitized patients, although the conventional serum analysis programs can identify included specificities to some degree, it is often difficult to be assured of the specificities. By development of computer programs that have the capability of storing the amino acid sequences for each of the panel cells, it has been possible to identit~¢ many high frequency sera 'as being directed to a "logical" epitope, represented by a single amino acid. We examined sera from 20 patients who had rejected their transplants. In one example, the kidney donor ~as A1 A24 B7 B62 and the recipient was A3 B7 B62. Thrce months after rejection, the antibody present in the recipient was anti A1,2,24,25,26,31,32,and 33. The program identified this antibody as simply being one directed against histidinc at positon 70. In this way, many "multispecific" antisera can be calssified more simply classified according to the amino acid 'epitope' against which they are directed. Moreover, the epitopes which result after rejection of a transplant can be postulated as the important immunogenic epitopcs against which the recipient reacted. To date, because of the multispecificity, sera from patients who had rejected have been difficult to analyze. Many more sera from patients who had rejected grafts are being investigated, and the program is being extended to analyzc the Class II specificities.