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patient paris
82
Abstracts
P442
P441
HIGH RESOlUTION HLA MATCHING ASSOCIATED WITH DECREASED MORTALITY AFTER .
UNRELATED BONE MARROW TRANSPLANTATION. Jeannet MieIlel' , Speiser Daniel', Rufer Nathalie', GrundschoberChrislop/le', GralWohl AJois', Chapuis Bernard', HeIg Claudine', LiiligerCornelius', S'enMarja-Kaisa', Roosnek Eddy', and TIerC)' Jean-Mariel . 'Department ofInternalMedicine, UniversityHospital, Geneva, Switzerland, 'Di~s ion ofHematology, University Hospital Basel, Switzerland, ' University Hcspital Eppendorf, Hamburg, Germany and ' UniversityCentral Hospital, Heisinki, Finland As comparedwilh related HLA identicalsiblingdonors, bone marrow Iran'planlation (BMT) with phenotypically HLA ABDR-'tompaUble' unrelated donorsis associated withincrea' edmalta/ity. This maybedue 10 HLAinrompalibilifa nol detected byron,entlana! typing. We have analyzed44 unrelaledpaUBnt-OOnor pairswho weramaicIled ~ HLA-A,-B, and-OR byrOUbne tissue typing. 0 ... canprehensiYe HLA typing approach consisled 01serology,cytlltoxic T cell precursor (CTLp) tests, Tcell doning, oIigolyping and DNAsequencing. This allowed the idenli1icatioo of numerous HLA mismalchesmissed by!herouline typing procedure. 24 patienl-donoc pairs were highly matched, whilethe remai1ing 20 pairs were allele mismatched at ertherHLA·A.-B,-C,-DR, or -DO loci, andIor for !heCTLp lest Patient and donor age, di....,osis, and treatment
p443
BoNEMARROWlllANSPLAHTATlON (8MT) WITH UNRELATED DONORS IN NORWAY
Egela'ld Tastei1, 8l'i1d1 L.aentz. EYellSBO Steil A AlTed1lsen Dagfi1n, 'll1cxsl1t E, Institute of Tra nsplantation Immuno logy, Med~ Depa rtment A, and Surgical Departm ent B, The National Hospital , Oslo , Norway In 1990 - 1995, 32 Norwegian adult patients, mean age 31 years (15-50), 16/14 males/females , underwent BMT with unrelated donors . 21 patients had CML in CP, 6 had AML in 1. or 2, rem, and 5 had ALL In 2. or 3. rem. 10/13 CMV neg patients received CMV neg marrow , Busulphan , 16 mglkg , and cyclophosph amide , 120 mgl kg, were used for cond itioning. Standard cyc1osporin N melothrexate acute graft versus host disease (AGVHD) prophylaxis was given. All but one donor matched serolog ically for HLA-A and -B and fo r DRBl and DOB1 by SSOP or DNA sequencing. 13/21 donors were mismatched for one DPB1 al ele. A compatible donor was found for 78% of tile patients. 25 patients were transplanted at Th e National Hospital : Fille patients (20%) developed AGVHD grade III-IV and 11 (44%) patients had AGVHD grade I-II. AGV HD was observed w~h and w~hout DPB1 mismatch es. One rejection was seen. 19 (76%) pal ients are alive 2-66 month s post-BMf. Five patients died within 8 months and one 17 months post-BMT. The data indicate that BMf with unrelated HLA-A , B, DRB1 , and DOB1 matched dono rs gives results approach ing those of BMT w~h HLA-identi cal sibs. Matching for DPB1 seems 10 be less important. BMT with unrelaled donors wilh one HLA-A, B, or DR mismatch is being considered to allow more patienls be lransplanted.
P445
MOLECULAR ANALYSIS OF CLASS U HLA DISPARITY IN BONE MARROW TRANSPLANTS USING UNRELATED DONORS Baxter-Lowe LA, Noreen H, Salazar M, Schmeckpeper B, Williams TM, Granja C, Awdeh Z, Begovich A, Kitajima D, Morse L, HeglandJ, Ng J, Hartzman R, Hurley C,Chapek M
National MarrowDonorProgram. Minne apol is, Minnesota, USA
Retrospective analysis of class U HLA disparity was performedfor l ,302 donorrecipient pairs from unrelated-donorbane marrow transplants performed between1988 and 1994. A combination of DNA-based meth od s was used to resolve class II HLA alleles. The observed mismatches between donor and recipientfollowing high resolution class II typing is summarized by locusbelow. Lo cus
1mJl1
1!B1!J
1lliM~
!lQAllli!lUDPAl
nflll
0/. Mislnatched O bservations
19.5 931
14.0 508
11.0 219
13.9 JJ8
86.9 735
0,4 259
18.8 483
47.8 1190
Pre liminary ana lysis of these typi ngs re vealed substan tial Glass II HLA d isparity between donors and rec ipients , primaril y attr ibutab le to the high ra te of HLA·DP dispa rity. Disp arity be tween donor and recipient for HLA·DRB an d HLA-DQ was less freq uent. Disparities w ithin HLA ·DRBI groups were most freq uently observed for DRB [· 11 (23%), DRBI · U8 (20% ), and DRB I4 04 ([7% ). Each mismatch w ithin a loc us was furt her defined to include analysis o f the struct ural and funct iona l changes in the HLA molecule and the type (conservative ve rsus no n-conservative ) of amino acid substitutio n. For insta nce, the most frequently mismatched res idues encod ed by the sec ond exon of DRB I were P71 and p86. Mismatches, definedat various levels (incl udin g sero logic , gene ric DNA-b ased. alle lic and amino acid) and corre lated with patien t outcome. will be used to investigate th e clinical relevance of class II HLA dispar ity for bone marrow tran splantation using unrelated donors. The study was sponsored by the N ational Marr ow Donor Prog ram
(Office of Naval Research Grant #NOOOI4-93-I-0658).
CORRELATION OF RISKOF ACUTEGRAFT-VERSUS-HOSTDISEASE (GVHD) WITH DONOR-RECIPIENTMISMATCHINGFOR HLA·D06 1 ALLELES IN UNRELATED DONOR(URD) TRANSPlAN TATION Petersdort EW, Longton G, Anasetti C, Mickelson E, SmithA, Martin P, Hansen J Fred Hutchinson CancerResearch Center and the Universityof Washington, Seattle, WA, USA We studied the effect of HLA-DOB1 allele mismatching on the development 01acute GVHDin 4B9patients who underwent marrowtnansplantation from HLA-A, B, DR serologically malched URDs for hemalologic malignancy at our center between May 1, 1985 and June 30, 1995. All patent s received methotrexale and cycJosporine as GVHD prophylaxis and unmanipulated (T cell replete) marrow. HLA-DRBl and DOBl alleles were typed by mclecular nnethods. Of the 469 donor-recipient pairs, 343 were DRBl and DOBl matched, 47 we", DRB1 matched but DOBl mismatched, 51 were DRBl mismatched but DOBl matched, and 28 were DRBl and DOBl mismatched. In a univariable Cox model, the relative risk (RR) 01grades III-IV acute GVHDfor DOBl mismatching among DRBl matches was 2.0 (95% CI: 1.3, 3.0; p=.OO2) and tor DOBl mismatching among DRB1 mismatches was 1.5 (95% CI: .78, 2.7; p; .24). The RR of DOBl mismatching when stratified on DRB1 matchstatus was 1.8 (95% CI: 1.2, 2.6; p; .D02) HLA-DOB1 allele mismatching is a risk factor tor acuteGVHDin unrelated marrowlransplantation, and henceHLA-DO functionsas a transplantation antigen.
P444 UNRELATED BMT IN HUNGARY, 4 YEARS EXl"ERJENCE E.GyM . E.Nyiri, B.Ko!szi". L.TImb", Gy.Pelr.lnyi Had. lnatinne of Kaematology. BLood Tramfusion and ImmllnOlogy. Budapest; "Szt. LWI6 Hospital, Budapest, Hungary Hungarian Bone Marrow Donor Regj ~try was estabLisbed in L99L with the aim at providing the basic cootact bel-.J the clinical BMT progt:lD1 and the world-wide organization. Unrelated donor sean:hes have been initiated for 3S patients (19 children , 16 adult» via EDS ccm pe ter liDk up since 1992 . Altogether 87 voJu.nteers from diffeml1 rountri.n were asbd 10 give blood for DR t)lPing >nd 42 for MLC and DNA typing. Serological, cellular (MLC.CT Lp) >nd molcculla biol"iPcal metlloplantr:ieh indicated BMT were: CML (0= 10), AML (0~ 1) , ALL (0=1). SAA (0= 1), F>nconi anaemia (0=2). All dnnor ·..cipient pairs wen: HLA·A!l,DR ,DQ identical. HLA-DPB" dirren:d in 5 from the 8 DPS " typed pain. 10 eacb case MLC _ negative in GVH directioD. but in 2 CUC'S were iDcorlclusive in HVO din."Clion. CfLp f""lUCDCie> were measured in 6 cases (2 low and 4 IOgll 1ev1:1l.C died sbonly after transplaDlation. Thi s worl< " .. supported in part by the Europe8Jl Foundation forScienees, Arts and Culture, Pari..Ams1erdam, by Hungarian MinilIuyofWellfan:(ETI-T227/1 993)
P446
HLA-Cw MISMATCHES IS PREDICIlVE OF B27-44 SUBTYPE MISMATCHESIN UNRELATED MARROWDONOR/PATIENTPAIRS. RAFFOUX C, TATAR! Z. FORTIER C, TAMOUZA C, MARZAIS F, O. CHARRON, Laboratoire d'lmmunologie et d'Histocornpatibilite, Hopital Saint Louis, Paris. France, HLA-C serologically defined alleles have been described in strong linkage disequilibrium with HLA-B alleles: Cw4 with B35. Cw8 with B14. In unrelated donor/recipient situations, we developed a PCR-RFLP Typing technique which is able to define 21 HLA-Cw alleles among the 36 defined by sequencing . We performedmolecular subtyping of B44 or B27 alleles using PCR/SSO and DGGE techniques (B4402,03,04,05,06/ 2701,02,03,04,05,06,07,08,09) in 14 pairs sharing these alleles among 100 donor/recipient pairsserologically HLA-AB identical
Cw identical Cw different
B27-844 identical 8
B27-B44 different
2
4
o
We suggest the possibility to use a new decisional tree for the choice of the unrelated donor: First, AB serological typing, then HLA ORBI, then Cw molecular typing followed among the Cw-ORBI identical donors to the patientby AB molecular genotyping,These results emphazise the importance of HLA Cw and AB molecular Typing in donor selection.
Abstracts
P442
P441
HIGH RESOlUTION HLA MATCHING ASSOCIATED WITH DECREASED MORTALITY AFTER .
UNRELATED BONE MARROW TRANSPLANTATION. Jeannet MieIlel' , Speiser Daniel', Rufer Nathalie', GrundschoberChrislop/le', GralWohl AJois', Chapuis Bernard', HeIg Claudine', LiiligerCornelius', S'enMarja-Kaisa', Roosnek Eddy', and TIerC)' Jean-Mariel . 'Department ofInternalMedicine, UniversityHospital, Geneva, Switzerland, 'Di~s ion ofHematology, University Hospital Basel, Switzerland, ' University Hcspital Eppendorf, Hamburg, Germany and ' UniversityCentral Hospital, Heisinki, Finland As comparedwilh related HLA identicalsiblingdonors, bone marrow Iran'planlation (BMT) with phenotypically HLA ABDR-'tompaUble' unrelated donorsis associated withincrea' edmalta/ity. This maybedue 10 HLAinrompalibilifa nol detected byron,entlana! typing. We have analyzed44 unrelaledpaUBnt-OOnor pairswho weramaicIled ~ HLA-A,-B, and-OR byrOUbne tissue typing. 0 ... canprehensiYe HLA typing approach consisled 01serology,cytlltoxic T cell precursor (CTLp) tests, Tcell doning, oIigolyping and DNAsequencing. This allowed the idenli1icatioo of numerous HLA mismalchesmissed by!herouline typing procedure. 24 patienl-donoc pairs were highly matched, whilethe remai1ing 20 pairs were allele mismatched at ertherHLA·A.-B,-C,-DR, or -DO loci, andIor for !heCTLp lest Patient and donor age, di....,osis, and treatment
p443
BoNEMARROWlllANSPLAHTATlON (8MT) WITH UNRELATED DONORS IN NORWAY
Egela'ld Tastei1, 8l'i1d1 L.aentz. EYellSBO Steil A AlTed1lsen Dagfi1n, 'll1cxsl1t E, Institute of Tra nsplantation Immuno logy, Med~ Depa rtment A, and Surgical Departm ent B, The National Hospital , Oslo , Norway In 1990 - 1995, 32 Norwegian adult patients, mean age 31 years (15-50), 16/14 males/females , underwent BMT with unrelated donors . 21 patients had CML in CP, 6 had AML in 1. or 2, rem, and 5 had ALL In 2. or 3. rem. 10/13 CMV neg patients received CMV neg marrow , Busulphan , 16 mglkg , and cyclophosph amide , 120 mgl kg, were used for cond itioning. Standard cyc1osporin N melothrexate acute graft versus host disease (AGVHD) prophylaxis was given. All but one donor matched serolog ically for HLA-A and -B and fo r DRBl and DOB1 by SSOP or DNA sequencing. 13/21 donors were mismatched for one DPB1 al ele. A compatible donor was found for 78% of tile patients. 25 patients were transplanted at Th e National Hospital : Fille patients (20%) developed AGVHD grade III-IV and 11 (44%) patients had AGVHD grade I-II. AGV HD was observed w~h and w~hout DPB1 mismatch es. One rejection was seen. 19 (76%) pal ients are alive 2-66 month s post-BMf. Five patients died within 8 months and one 17 months post-BMT. The data indicate that BMf with unrelated HLA-A , B, DRB1 , and DOB1 matched dono rs gives results approach ing those of BMT w~h HLA-identi cal sibs. Matching for DPB1 seems 10 be less important. BMT with unrelaled donors wilh one HLA-A, B, or DR mismatch is being considered to allow more patienls be lransplanted.
P445
MOLECULAR ANALYSIS OF CLASS U HLA DISPARITY IN BONE MARROW TRANSPLANTS USING UNRELATED DONORS Baxter-Lowe LA, Noreen H, Salazar M, Schmeckpeper B, Williams TM, Granja C, Awdeh Z, Begovich A, Kitajima D, Morse L, HeglandJ, Ng J, Hartzman R, Hurley C,Chapek M
National MarrowDonorProgram. Minne apol is, Minnesota, USA
Retrospective analysis of class U HLA disparity was performedfor l ,302 donorrecipient pairs from unrelated-donorbane marrow transplants performed between1988 and 1994. A combination of DNA-based meth od s was used to resolve class II HLA alleles. The observed mismatches between donor and recipientfollowing high resolution class II typing is summarized by locusbelow. Lo cus
1mJl1
1!B1!J
1lliM~
!lQAllli!lUDPAl
nflll
0/. Mislnatched O bservations
19.5 931
14.0 508
11.0 219
13.9 JJ8
86.9 735
0,4 259
18.8 483
47.8 1190
Pre liminary ana lysis of these typi ngs re vealed substan tial Glass II HLA d isparity between donors and rec ipients , primaril y attr ibutab le to the high ra te of HLA·DP dispa rity. Disp arity be tween donor and recipient for HLA·DRB an d HLA-DQ was less freq uent. Disparities w ithin HLA ·DRBI groups were most freq uently observed for DRB [· 11 (23%), DRBI · U8 (20% ), and DRB I4 04 ([7% ). Each mismatch w ithin a loc us was furt her defined to include analysis o f the struct ural and funct iona l changes in the HLA molecule and the type (conservative ve rsus no n-conservative ) of amino acid substitutio n. For insta nce, the most frequently mismatched res idues encod ed by the sec ond exon of DRB I were P71 and p86. Mismatches, definedat various levels (incl udin g sero logic , gene ric DNA-b ased. alle lic and amino acid) and corre lated with patien t outcome. will be used to investigate th e clinical relevance of class II HLA dispar ity for bone marrow tran splantation using unrelated donors. The study was sponsored by the N ational Marr ow Donor Prog ram
(Office of Naval Research Grant #NOOOI4-93-I-0658).
CORRELATION OF RISKOF ACUTEGRAFT-VERSUS-HOSTDISEASE (GVHD) WITH DONOR-RECIPIENTMISMATCHINGFOR HLA·D06 1 ALLELES IN UNRELATED DONOR(URD) TRANSPlAN TATION Petersdort EW, Longton G, Anasetti C, Mickelson E, SmithA, Martin P, Hansen J Fred Hutchinson CancerResearch Center and the Universityof Washington, Seattle, WA, USA We studied the effect of HLA-DOB1 allele mismatching on the development 01acute GVHDin 4B9patients who underwent marrowtnansplantation from HLA-A, B, DR serologically malched URDs for hemalologic malignancy at our center between May 1, 1985 and June 30, 1995. All patent s received methotrexale and cycJosporine as GVHD prophylaxis and unmanipulated (T cell replete) marrow. HLA-DRBl and DOBl alleles were typed by mclecular nnethods. Of the 469 donor-recipient pairs, 343 were DRBl and DOBl matched, 47 we", DRB1 matched but DOBl mismatched, 51 were DRBl mismatched but DOBl matched, and 28 were DRBl and DOBl mismatched. In a univariable Cox model, the relative risk (RR) 01grades III-IV acute GVHDfor DOBl mismatching among DRBl matches was 2.0 (95% CI: 1.3, 3.0; p=.OO2) and tor DOBl mismatching among DRB1 mismatches was 1.5 (95% CI: .78, 2.7; p; .24). The RR of DOBl mismatching when stratified on DRB1 matchstatus was 1.8 (95% CI: 1.2, 2.6; p; .D02) HLA-DOB1 allele mismatching is a risk factor tor acuteGVHDin unrelated marrowlransplantation, and henceHLA-DO functionsas a transplantation antigen.
P444 UNRELATED BMT IN HUNGARY, 4 YEARS EXl"ERJENCE E.GyM . E.Nyiri, B.Ko
P446
HLA-Cw MISMATCHES IS PREDICIlVE OF B27-44 SUBTYPE MISMATCHESIN UNRELATED MARROWDONOR/PATIENTPAIRS. RAFFOUX C, TATAR! Z. FORTIER C, TAMOUZA C, MARZAIS F, O. CHARRON, Laboratoire d'lmmunologie et d'Histocornpatibilite, Hopital Saint Louis, Paris. France, HLA-C serologically defined alleles have been described in strong linkage disequilibrium with HLA-B alleles: Cw4 with B35. Cw8 with B14. In unrelated donor/recipient situations, we developed a PCR-RFLP Typing technique which is able to define 21 HLA-Cw alleles among the 36 defined by sequencing . We performedmolecular subtyping of B44 or B27 alleles using PCR/SSO and DGGE techniques (B4402,03,04,05,06/ 2701,02,03,04,05,06,07,08,09) in 14 pairs sharing these alleles among 100 donor/recipient pairsserologically HLA-AB identical
Cw identical Cw different
B27-844 identical 8
B27-B44 different
2
4
o
We suggest the possibility to use a new decisional tree for the choice of the unrelated donor: First, AB serological typing, then HLA ORBI, then Cw molecular typing followed among the Cw-ORBI identical donors to the patientby AB molecular genotyping,These results emphazise the importance of HLA Cw and AB molecular Typing in donor selection.