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PRINCIPLES OF HLA TYPING; HLA MATCHING IN HSCT David Smillie H & I, NHSBT, Sheffield Histocompatibility & Immunogenetics Blood and Transplant • successful HSCT depends on many factors (disease, stage, age, treatment regime etc) • not least is HLA compatibility between patient and donor! Histocompatibility & Immunogenetics Blood and Transplant HLA TYPING REPORT – a collection of letters and numbers, how we arrive at this and what use is it? Histocompatibility & Immunogenetics Blood and Transplant DEFINITIONS • HLA = Human Leucocyte Antigen • membrane glycoproteins on all nucleated cells • ‘classical’ HLA loci, Class I (A,B,C) & Class II (DR,DQ,DP) each encoded by separate genes • recognised by the immune system as ‘self’ or ‘non self’ • this determines histocompatibility = acceptance/rejection of foreign tissue (e.g transplant) - host vs graft, graft vs host, graft vs leukaemia) • cellular immunity • antibody response • most polymorphic system in human genome - challenges for HLA typing and donor selection! Histocompatibility & Immunogenetics Blood and Transplant AMINO ACID POLYMORPHISM (this is what the immune system recognises) HLA molecule e.g HLA-A1 Histocompatibility & Immunogenetics e.g HLA-A2 Blood and Transplant DNA POLYMORPHISM (resolved by DNA typing) • SNP = Single Nucleotide Polymorphism • alleles differ by or more SNP Histocompatibility & Immunogenetics Blood and Transplant HLA ANTIGENS ON NUCLEATED CELLS DR DQ C DP B A DP A B DQ DR Histocompatibility & Immunogenetics paternal haplotype maternal haplotype C Blood and Transplant INHERITANCE OF HLA HAPLOTYPES Father + Mother = haplotypes (25% chance of identical sib) PARENTS A* a b c d CHILDREN a/c a/d b/c b/d Histocompatibility & Immunogenetics b/r B* C* DRB1* DQB1* (a) 01 08 07 03 02 (b) 03 07 07 15 06 (c) 02 44 05 04 03 (d) 30 13 06 10 05 (r) 02 44 05 10 05 Blood and Transplant ORGANISATION OF HLA GENES CHROMOSOME Histocompatibility & Immunogenetics Blood and Transplant HLA TYPING METHODS 1950’s 1960’s 1980’s 1990’s 1999 2000 2000’s discovery of HLA system serological typing first HLA genes cloned, sequenced DNA/PCR based HLA typing sequence entire MHC (HGP) database of all HLA alleles SBT, Luminex SSO Histocompatibility & Immunogenetics Blood and Transplant PROBLEMS ASSOCIATED WITH UNRELATED DONOR SEARCHING problems are: incomplete registry data (e.g no HLA-C or DQB1) HLA polymorphism (only 40-50% Caucasians have 10/10 HLA match, other groups less) • rare alleles/allelic variants • ethnicity • linkage disequilibrium donor drop out Histocompatibility & Immunogenetics Blood and Transplant PROBLEMS ASSOCIATED WITH UNRELATED DONOR SEARCHING (1) incomplete registry data • HLA • not all donors typed by DNA techniques • not all donors typed for DRB1 • not all donors typed for C &/or DQB1 • very few donors high resolution typing • gender, blood group, ethnicity, CMV not always available • costs Histocompatibility & Immunogenetics Blood and Transplant PROBLEMS ASSOCIATED WITH UNRELATED DONOR SEARCHING (2) HLA polymorphism • rare alleles/allelic variants (5,880 Class I, 1647 Class II alleles) • linkage disequilibrium Histocompatibility & Immunogenetics Blood and Transplant NO 10/10 DONOR BECAUSE OF RARE ALLELES HLA: A* B* C* DRB1* DQB1* JM 02:0 03:0 07:0 40:0 02:0 07:0 13:01 14:01/54 05:0 06:03 DEDKM 2127057 02:0 03:0 07:0 40:0 02:0 07:0 13:01 14:01/54 05:0 06 02#1 - 07 40:0 02 07 13:01 14 05 06 02#1 03 07 40:0 03 07 13:01 14 05 06 GB 1300733 DEDKM 620547 #1 Not HLA A*02:05 Histocompatibility & Immunogenetics Blood and Transplant NO 10/10 DONOR BECAUSE OF RARE ALLELES MUD’s HLA: A* B* C* DRB1* DQB1* SH 03 24 15:18 18 05 07 04:07 13:01 03:01 06 TO03 3992 01 24 15:10 18 03 12:03/06 04:02 11 03:02 05 AKB-142342 24 25 15:18 18 07 12:03/06 04:01 13:01 03:02 06 CBU’s (matching for HLA-A, B & DRB1 only) HLA: Panel SH N/A 6504125 A* B* C* DRB1* DQB1* Vol (ml) TNC (107) 03:01 24:02 15:18 18:01 05:01 07:04 04:07 13:01 03:01 06 N/A N/A ACCB 03 24:02 38:01 35:23 07:02 12:03 04:07 13:01 03:02 06:03 121 318 999293104 UICB 03 24 35 - 04 04:07 13:01 N/T 30 208 290055467 AUCB 03 24 27 35 04:07 13:01 N/T 60 91.9 Histocompatibility & Immunogenetics N/T Blood and Transplant SUITABLE DONOR DESPITE RARE ALLELES HLA: A* B* C* 02:11 11:01 35:03 40:06 04:0 DEDKM 3571314 02:11 11:01 35:03 40:06 04:0 15:02 0564-3760-1 02:11 11:01 35:03 40:06 12:03 15:02 DM Histocompatibility & Immunogenetics 15:02 DRB1* DQB1* 10:01 15:01 05:01 06:01 10:01 15:01 05:01 06:01 10:01 15:01 05:01 06:01 Blood and Transplant LINKAGE DISEQUILIBRIUM • some alleles occur more frequently together than expected by random association • extended (ancestral) haplotypes e.g.A*01, B*08, C*07, DRB1*03, DQB1*02 • commonly found HLA-B & C, HLA-DRB1 & DQB1 • patients with common HLA-B and -C or HLA-DRB1 and -DQB1 associations have a positive impact on the likelihood of finding a donor • patients with uncommon HLA-B and -C or HLA-DRB1 and -DQB1 associations have a negative impact on the likelihood of finding a donor Histocompatibility & Immunogenetics Blood and Transplant LINKAGE DISEQUILIBRIUM HLA-B & C Histocompatibility & Immunogenetics Blood and Transplant LINKAGE DISEQUILIBRIUM HLA-DRB1 & DQB1 Histocompatibility & Immunogenetics Blood and Transplant COMPOUNDING EFFECT OF LINKAGE DISEQUILIBRIUM A* HLA: B* DRB1* C* KaM 02:0 68:01 27:0 44:0 02 07 GB 1545503 (AN) 02:0 68:01 27:0 44:0 02 07 02 68:0 27 44 02 05 DE-BBB 12355 02 68:01 27 44 05 DE-DKM 336800 02 68 27 44 DE-DKM 513225 02 68:01 27 DE-DKM 2473710 02 68:01 0213-6872-5 02 0491-2420-9 02 1/026701 (BBMR) DQB1* 08:0 13:02 03 06:04 11:01 13:02 03 06:04 08 13:01 04 06:03 - 08 13:01 03 06:03 01 07 08 13:02 04 06:04 44 01 05 08 13:01 04 06:03 27 44 05 07 08 13:01 04 06:03 68:01 27 44 03 07 08 13:02 04 06:04 68 27 44 01 07 08 13:02 04 06:04 Histocompatibility & Immunogenetics Blood and Transplant WHAT IS THE RISK OF HLA MISMATCHING ? • graft failure (rejection) • GVHD (but GVL; ?HLA-DPB1) • selecting a mismatch at locus may affect other loci due to linkage disequilibrium Histocompatibility & Immunogenetics Blood and Transplant 16th IHW PROJECT INTO HLA MISMATCHING Histocompatibility & Immunogenetics Blood and Transplant 16th IHW PROJECT - RESULTS Histocompatibility & Immunogenetics Blood and Transplant HLA MISMATCHING – NO CONSENSUS IN UK! Histocompatibility & Immunogenetics Blood and Transplant HLA NOMENCLATURE Histocompatibility & Immunogenetics Blood and Transplant