382 Annual AACHT Mecun~ 1981

cel ls with appropriate HLA phenotypes. The Ql/28 antigenic determinant is ,e~arkabiy resistant to denaturing treatments and Is present on both fr.-e and ~2-~nicroglobuiln assoc,ated HLA-A,B heavy chain, therefore definsng ~or the f i r s t time conformatlonal s ;mi lar i t ies between the two states of HLAoA,B heavy chain. Both Q|/28 and (~/~/* determinants are of protein nature, since they disappear upon pronase digestion of 3H-glucosamlne labeled material. However, the expression o f the determinant reactive with HoAb Q~/6~ is dependent on the presence of the carbohydrate moiety, because the HoAb Q~/~I, is unreactive with HLA-A,6 antigens syntheSiZed by lymphoid cel ls In presence of tun;camycin.

FIVE HIA-D ANTIGENIC CLUSTERS ASSOCIATED VITH IlIA-DR4. Bancy.Resnsmoen~ Hsrsam Sessl l and F r i t z H. Bach from th~ lmaunobsolo~y Research Center, Unxvers l ty-of Hsnaesots, Hxnneapolss, HN 55455

Our recent studies have confsrmed the ezssteace of four IIlA-D c lus te rs assoclated with HLA-DR4: Dw4, l)vlO. DBJ, DIT; and have Idrn t l f lod a new antisenxc c lus te r . LDAO. Fairy-seven DR4 posltxve respoaders ~ere tes ted wsth twenty RLA-DR4 assoczated homozygous typsnJ; c e l l s (I/TCs) representins these fzve ant%genlc clu~ters and also the ST-2 c lus t e r . Dr4 was defined by 5 HTCs and DviO by 4. The spectfsclty D~ was defined by 8W402 (TsnJ~t) and • local HTC, Dlfas A new specificity provsssonally designated LD~O was defsned by four RTCs: 15-40 ( loca l ) , ~ (~anaen), JlZou (Hsu) and 8~rJ09 (Sucsu-Foca). In a total of 97 sndsvsduals, DP~ was assocsated wxth I~4 xn 45~ of cases and wsth Dwl0 in I$~; of those DR4 hsplotypes not associated wltb Dw4 or Dw)O, 15 (16~ of the t o t a l ) were LI)~O, 9 (9~) were DBJ, 3 (3~) were DYI, and 12 (12~) ~ r e blank.

The Eishth Jnternatsonal Workshop Identlfs~d two HTC~, 8V322 (Festenstexn/Sachs) end 81/318 (Grosne-Vllde). as recognlz~ng • new DR~- associated specif ici ty, DBJ; however, our s tudies ladlc~te th i s c lu s t e r ss complex and appears to give typing response pat terns overlappsng wsth the I~r-2 c lu s t e r , as defined in our s tudies by three Japanese IITC~ (ST-2. HI, KA-TsuJl), and with a broader c lu s t e r defined by th~ RTCs DSto (Rsu) sad REd~ (Yestenstein/Sacbe ).

These studies are informative not only in defining the l)R&-assoelsted ~.LA-D c lus t e r s , and in s~q~orttng the concept tbe t D sad DR cannot be con° stdered sdentscal , but a lso in e~Fhaslzjn z the complexity of the ]/IA-D tea/on.

FUR33~R ~ALYSIS OF D-~GION CO~I~XIIT WITH YLT CLO~T~o ~ancy~ tns~e_n z Nat sam Se|a.ll and F r i t z H . . h c h , from the Ism~nobiolo~ Research Center, Unsver- s l t y of Hsnnesota. Hinneapolis, tin 554.55.

We have i n v e s t l p t e d pat terns of r e a c t l v l t y of "clones" obtslned from bulk PLT reacents. One reagent was generated a£asnst an entsre haplotype ussng fomtlial haplosdentsc~ c e l l s for pr/msnS. 128 "clones" were screened on day 20; 2°$-$,0 x I0 respeadlns c e l l s frequently save reactsvstses sn excess of I00,0OO-¢pm. with controls less th~n I000 ~m. Of the "clones" . 80~ were PLT-reactsve. soclndlns D*spectflc. s t imulator- specular, and DR-speclfsc pat terns of r eac t ivs ty , aa well aa several "broad" pat terns . The bulk PLT shoved Jtronll res t l smlat lon associated with IILA-D and not DR; sn p a r a l l e l . D-speclf lc clones were much so t , frequent than DR-specsfsc ones, ~lhereas , r / s i n g in a D/DR zncomp;tlble colblnat lon y le lds many clones vsth reactivsties associated vsth D and DR, fur ther analysts of the D region i s s imphf ted i f there i s D/~R matchsng sn the prmlng combsnatson, as i l l u s t r a t e d by another bulk PLT. "his reagent was generated between two fomt l i s l , beploidentsc~l c e l l s phenosdentscai for the D/DR specsfJcitleS, and appeared to de tec t determ,nants not assocsatcd with the s p e c i f s c i t l e a defined by conven- tsonal D/DR typsns. We obtained tvo major patterns of react iv i ty su Lhe "clones" from thss bulk reagent; both were unrelated to D or DR specsf t -

Abstracts .~8.~

c i t i e s o f the res tzmula tzng c e l l s Although the most f requent ps t t e rm of react iv i ty was associated ~zth SB2, clones uhich ~ere associated ~zth n e i t h e r D, DR, nor SB were a l so zdentifsed. The model o f the HLA-D regxon that can be derived from these studies, as I t re lates to overal l D-region complextty and to test ing ~zth ants-DR sera and HTCs, ~s l | be discussed.

PHYTOHEPiAGGLUTININ P STIMULATED HUMAN LYMPHOCYTES FOR RAPID TRANSPcANT ~ATCHING. F. Robbln% R.3 Hartzman, [mnunoloq~c Oncoloav O~ws~on, Georqetnw~ University School of Hedtcine~ gashlnqton, O.C. 20007 and the Tissue Bank, Naval qedical Research Ins t i tu te , ~ethesda, ~D ~0014.

Th~s research concerns the development of a rapid cross-matchlna procedure for transplantation. Human lyhmphocytes, ten days after treatment with phytohemagglut~n~n P /PHA-P), show PLT (secondar~ ~n v i t r o restlmulatlon) reaction patterns which correspond to the i r ce l lu la r tHLA-DI tYPe. Cymphocytes with shared ce l lu la r type stimulate PHA primed ce l ls to a lesser deqree wh~le the ma3or~ty of lwphocytes w~th unshared ~ LA-D type stimulate a s~qn~f~cantlv 9rearer response. Using a standard H thymd~ne ~ncorporat~on technique, PHA-PLT responses can be detected

as ear ly as I2 hours total ~ncubat~on t~me. Th~s method ~s simple, gives results which correspond to the primary mxed leukocyte response and thus may be useful as a cross-matching technique for cadaverlc renal transplantation.

PLT Stimulators TOI-FHA 101 94 NABA 523 68 127 20

DwS/? OwS/5 0w515 Ow~/5 Ow~l ~ Ow317 Dw41Q 24-36 hrs** 42* 533 578 625 1346 4130 400~ 48-60 hrs ~1 2153 2915 1161 ~968 9 8 1 8 14F2q

* ~PH 3H Thymldlne Incorporation ** ~H Thymldlne lahel~ng-harvest~nq times

HLA STUDIES REVEAL POSSIBLE NEW HUMAN T/ t VARIANT. P . J . Romano~ C. HeEwen~ T.O. Brewster I D.P. Bobeyt Foundation fo r Blood Research. Scarborou~h, HaIn~; Besiona~ memorial Hospital, Brunswick, Hazne, Sidney Farber Cancer Center, Boston, Hassachuse t t s

Cl~oxcal and Immunogenetxc s t u d i e s have been performed on members of 5 gene ra t i ons of a family tn which f a m i l i a l precocxous a r t h r i t i s has been r e p o r t e d . This dxsease zs c h a r a c t e r i z e d c l i n i c a l l y by the occurrence of a b n o r m l l y severe o s t e o a r t h r z t z s a t an ea r ly age, in add i t i on to deve lop- mental d e f e c t s such as sho r t e x t r e m i t i e s and stubby f sn~e r s . C l i n i c a l

I symptoms begin a t age 20-30 y e a r s .

Genetic and c l i n i ca l analyses strongly indicate dominant transmission of t h i s precocious a r t h r i t i s through 5 generations although HLA typinE could only be performed on members of 3 g e n e r a t i o n s . Seven of 0 ind iv idua l s in 2 gene ra t i ons have shown the d i s e a s e . Ten c h i l d r e n zn the | a s t gene ra t ion a re s t l ] l under age 20. Inx tza l HLA typ ing performed on these 0 ind iv idua l s has demonstrated t h e A2, BS, and/or A2, BI5 hap lo type(s ) zn the 7 a f f e c t e d family members. Pre l iminary haplotype segrega t ion ana ly s i s sn szblsn~ p a i r s f avo r s lankage of t h i s d i s e a s e wi th HLA.

Precocious o s t e o a r t h r z t i s zs a dxsease c h a r a c t e r i z e d by developmental de - f e c t s , possibly under genetic control of the human ma3or hzstocompatzbzlsty comp ex. Addi t ional family s t u d i e s w i l l be required t o a s c e r t a i n whether t h i s d i s e a s e ss c o n t r o l l e d by a gene t i c d e f e c t zn a region analogous t o t h e mouse T / t locus and l inked t o t he HLA region in humans.