Hyperacute renal allograft rejection in HLA-AB identical situations
2
Seventh Annual Meeting of the American Association for Clinical Histocompatibility Testing Abstracts HONOCLONAL ANTIBODIES TO HLMAH Ia GL¥COPROTEINS:THEIR REACTIVITY WITH B-LYHPHOCYTIC LEUKEHIC CELLS (CLL). JANE ADDISt HICHELLE LETARTE AND JUDY FAL~ HOSPITAL FOR SIC)( CHILDREN AND TORONTO WESTERN HOSPITALt TORONTO. Five hybridoma lines producing anCibodles to human Ia antigens have been established. Hice of haplotype u vere lumunlzed vlth la glycoproteins obtained from tanrocholate extracts of human B-CLL cells and fractionated by Lena culinerls chromatography and gel filtration. The monoclonal antibodies I-'~-~,' 18c2, '1'8d$. 21~ and 21r5 vere shm~n to be la specific by cellular radlo/amunoassay, cytotoxicity assay and inmunoprecipJtatlon. Strou K binding to B-CLL cells and B-lymphoblaatold cell ixnes yes observed; T-ALL and T cell lines did not bind significant amounts of antibodies. The five monoclonal antibodies vere of IsGI subclass and shoved complement-dependent cytotoxlcity only in the presence of rabbit anti-uouse IgO serum; normal B cells and B-CLL cells were lysed ~hile normal T cells vere not. Polyacryla- mlde gel electrophoresle revealed that the monoclonal antibodies /zmunopre- clpltated a and B chains and in some cases the invarlant chain of In. The monoclonal antibodies recognize nonpolymorphlc determinants of h~man la molecules. Binding. under saturating conditions, of the various monoclunal antlbodi~s to B-CLI, cells derived from 6 different patients revealed that smae of the antibodies vere reactive only vlth subpopulat£ons of human Ia molecules. B-CLL provide a unique source of cells, relatlvely homogeneous and rich in Za. The expression of Ia molecules on these cells, as measured by interaction vith several nonoclonal antibodies, can reveal subpopulatious of Is molecules not apparent from studies of normal cells alone. RENAL AI, LOGRAFT REJECTION IN IILA-AB IDENTICAL SITUATIONS. A. Ahern~ S . B . ATt ruc, P. DellaPelle, A.B. Cosini~ R.B. Colvin and T.C. Fuller. Dept. o£ Surlery an4 Patbolol~r, Harvard Hadtcal School eg the Hessachusetts General Hospital, l~stoa. Xt is considered that only T cell reactive alloantxbody is capable cf medimtin 8 classical hyperacute rejection of n renal silo|raft. TVo pationts, both demonatcet~n8 high deacees o£ T cell panel antibody, received ELA-AB identical0 -DR dissimilar aUografts (one living related, one cadaver) v~ch the donor T cell crosshatch (Amos and sntiglobulin cytotoxintty) v*th 811 esra yes negative. Further studies revealed that both patients had ktaho tttered (1:32-1:64) donor-specifxc B cell antibodies which vere reactive Hck122-26 March 1981 m Chrlando,Florida, USA Human lmmuno/oSy3. 351-394 (1981) O I~hcvJcr Nocsh Holland. lnc. 1981 52 Vandc/bd¢Ave. New York, FIY 10017 351 0198-8859/8 i/08351-4452 50
Transcript of Hyperacute renal allograft rejection in HLA-AB identical situations
Seventh Annual Meeting of the American Association for Clinical Histocompatibility Testing
Abstracts
HONOCLONAL ANTIBODIES TO HLMAH I a GL¥COPROTEINS:THEIR REACTIVITY WITH B-LYHPHOCYTIC LEUKEHIC CELLS (CLL). JANE ADDIS t HICHELLE LETARTE AND JUDY FAL~ HOSPITAL FOR SIC)( CHILDREN AND TORONTO WESTERN HOSPITALt TORONTO.
Five hybridoma l i n e s producing anCibodles to human Ia a n t i g e n s have been e s t a b l i s h e d . Hice of hap lo type u v e r e lumunlzed v l t h l a g l y c o p r o t e i n s o b t a i n e d from t a n r o c h o l a t e e x t r a c t s o f human B-CLL c e l l s and f r a c t i o n a t e d by Lena c u l i n e r l s chromatography and g e l f i l t r a t i o n . The monoclonal a n t i b o d i e s I-'~-~,' 18c2, '1'8d$. 2 1 ~ and 21r5 v e r e shm~n t o be l a s p e c i f i c by c e l l u l a r rad lo /amunoassay , c y t o t o x i c i t y a s say and i n m u n o p r e c i p J t a t l o n . S t rou K b i n d i n g t o B-CLL c e l l s and B- lymphoblaa to ld c e l l i xnes y e s obse rved ; T-ALL and T c e l l l i n e s d i d n o t b ind s i g n i f i c a n t amounts of a n t i b o d i e s . The f i v e monoclonal a n t i b o d i e s v e r e o f IsGI s u b c l a s s and shoved complement-dependent c y t o t o x l c i t y on ly i n the p re sence o f r a b b i t a n t i - u o u s e IgO serum; normal B c e l l s and B-CLL c e l l s were lysed ~ h i l e normal T c e l l s v e r e n o t . P o l y a c r y l a - mlde g e l e l e c t r o p h o r e s l e r e v e a l e d t h a t t h e monoclonal a n t i b o d i e s /zmunopre- c l p l t a t e d a and B c h a i n s and i n some case s the i n v a r l a n t c h a i n o f I n . The monoclonal a n t i b o d i e s r ecogn i ze nonpolymorphlc d e t e r m i n a n t s o f h~man l a mo lecu le s . B ind ing . under s a t u r a t i n g c o n d i t i o n s , o f t h e v a r i o u s monoclunal a n t l b o d i ~ s t o B-CLI, c e l l s d e r i v e d from 6 d i f f e r e n t p a t i e n t s r e v e a l e d t h a t smae of t h e a n t i b o d i e s v e r e r e a c t i v e on ly v l t h subpopula t£ons o f human I a molecu les . B-CLL p rov ide a unique source o f c e l l s , r e l a t l v e l y homogeneous and r i c h i n Za. The e x p r e s s i o n o f I a molecules on t he se c e l l s , a s measured by i n t e r a c t i o n v i t h s e v e r a l n o n o c l o n a l a n t i b o d i e s , can r e v e a l subpopu la t ious o f I s molecules no t a p p a r e n t from s t u d i e s o f normal c e l l s a l o n e .
RENAL AI, LOGRAFT REJECTION IN IILA-AB IDENTICAL SITUATIONS. A. Ahern~ S.B. ATt ruc , P. D e l l a P e l l e , A.B. Cosini~ R.B. Colv in and T.C. F u l l e r . Dept . o£ S u r l e r y an4 Patbolol~r, Harvard Hadtcal School eg the Hessachusetts General Hosp i ta l , l~s toa .
Xt i s considered tha t only T c e l l reac t i ve a l loantxbody is capable c f medimtin 8 c l a s s i c a l hype racu te r e j e c t i o n o f n r e n a l s i l o | r a f t . TVo p a t i o n t s , b o t h demonatcet~n8 h i g h deacees o£ T c e l l pane l a n t i b o d y , r e ce ived ELA-AB i d e n t i c a l 0 -DR d i s s i m i l a r a U o g r a f t s (one l i v i n g r e l a t e d , one cadaver ) v ~ c h t h e donor T c e l l c r o s s h a t c h (Amos and s n t i g l o b u l i n c y t o t o x i n t t y ) v * t h 811 e s r a yes n e g a t i v e . F u r t h e r s t u d i e s r e v e a l e d t h a t b o t h p a t i e n t s had k taho t t t e r e d ( 1 : 3 2 - 1 : 6 4 ) d o n o r - s p e c i f x c B c e l l a n t i b o d i e s which v e r e r e a c t i v e
Hck122-26 March 1981 m Chrlando, Florida, USA
Human lmmuno/oSy 3. 351-394 (1981) O I~hcvJcr Nocsh Holland. lnc. 1981 52 Vandc/bd¢ Ave. New York, FIY 10017
351 0198-8859/8 i/08351-4452 50
352 Annual AA(.HT Meetmg, 1981
a t a l l tewperatures but were not au to- reac t lve or plat~elet adsorbable. An /smunofluorescence complement f ~ a t l o n assay with sere from the l lvzn 8 re la ted rec ip ien t and donor skin as substrata demonstrated br lght s t e ln log o f the d e m l mlcrovssculatere; th i s antibody i s adsorbable v / t h Al,Ba,DBv3 HTC LCL. In an addi t ional s e r i e s o f 17 b i l h l y sens i t i zed r ec ip i en t s , another case of pre- t ransplant h l~h- t l t e red donor - s l~c i f l c B c e l l antibody was found. TbLs slIol;raft wan lost (23 days) due to acute accelerated vascular r e j ec t i on , in comparlno~ to the 16 B c e l l crossmai~h negatlve pa t i en t s in whom no compromzse of eor ly functzon was observed. Prolbelnary analysts IndLcat~es that these B c e i l r eac t ive ant lbodles are not d i rec ted against the HLA-DR detemlnanta . Our studxes a n n e n t tha t h igh - t l t e r ed donor B c e l l antibody reac t ive with vascular endotheltum presents an Insumunntable b a r r i e r to successful r e ~ l t ransp lan ta t ion .
HLA TYPING OF HONOCYTES. G- A~b~ P.I. Terasakfe and H. Haktmoto. Tissue Typing Laboratory, UCLA School of Hedicine, University of California, Los A~eles, COlIf.
Honocytes can now be prepared in highly purif ied suspensions verifiable by the Coulter counter. Using such preparations, we have reotnvesttgated HLA typing of monocytes. Although monocytes are often hard to k111 in the cyto- tox ic i ty test, they react to HLA-A,B,C sere In exactly the same way as T lym- phacytes. That Is, they can be typed for these antigens using the same Corn* plement and 1~ hr incubation period, indicating that the eonocytes are sus- ceFttble to lysts. For HLA-DR typing, mnocytes require an extra hour of in- cubation but othenvtse tend to have the same antigens as B lymphocytes (Table 1). The HT1 to HT4 sere tend to react to a s l ight ly lesser extent with mono- cytes than with B lymphocytes. Honocytes do not react with the sere defining Tel, 2. 3, or 4. Although Lewis I or Lewis a reacted In 811 positive cases wtth the monocytes as well as B lymphocytes, Leads 2 and Lewis S did not re- act with monocytes. I t is also of interest that 8 c~ld antibodies did not react wtth monocytes.
We conclude that the peculiarit ies of mnocyte t~plng as compared to 8 cel l typing are part ia l ly attributable to a general weakness of reactions with monocytes but that there are also qualitative differences tn the d is t r i - bution of antigens on these cells as cmpared to B ly~pbocytes.
CHARACTERIZATION OF CYTOTOXIC LYHI~OCYTES AGAINST t~.A-A,B,G AND HLA-D/DR ANTIGENS. Edward Ball t Jun Okada e and Peter Stastny. Department of Internal I~Jtcfne, The University of Texas Health Science Center at Dallas, lexes. . .
CytotoxIc lymphocytes specific for HLA-D/D~ on monocyr~s or B lympnocytes have been generated with stimulating cel ls atsmtched only at the HLA-D re- gton (J. Exp. Had. 149:485, 1979). In the present ~ork we have gee~'ated effector cel ls against products of malttple HI.A loci and analyzed thin singly on selected target cel ls expressing only one of the relevant antigens. Thts pemltted comparison of Class I andClass I I HUt antigens as targets. using the same effector pepulattons. A stngle HLA-A or B target antigen produced 41,6 percent lysts, a stngle D/DR detemtnant, 39*5 percent. The effector cel ls were studied ustng monoclonal antibodies: OKT3, that reacts with most peripheral bloud T cel ls ; OKT4 and OKT8 spac|ftc for markers of T cell subsets (Reinherz et e l , 1979). Plixed 1)mpbocyte culture-activated cells were treated with mnoclonol antibody and c~mplemant prior to the cyto- tox ic i ty assay. OKT3 reduced cytotoxtc effector act iv i ty by 87t7 percent for Class I and 81,1 percent for Class Yl targets, indicating that the k i l lers are T l~nphocytas tn both cases. CytotoxIc cel ls for HIA-A,6 antigens were mostly T8% whereas ktl le~s agatest HLA-D/DR were reduced as effect ively by anti-T4 (62*7 percent Inhibit ion of lysts) as by . antt-T8 (40*7 percent inhibi t ion). The difference bet~en anti-class I --no anti-class I I k t l le rs , with regard to Inhibit ion by OKT4 antibody and co , leo ment. was highly signif icant (1~0.0001). The results indicate that cytotoxtc T cel ls recognizing HLA-D/OR 8re produced together with those recognizing HLA-A,B,C antigens, but that they are di f ferent on the has|s of surface phenotyplc markers as well as target antlgen specif ic i ty.
