Transcript duquesnoy08efi
Clinical Relevance of HLA Epitope Antigenicity and Immunogenicity Rene Duquesnoy University of Pittsburgh Medical Center
Matching: The Traditional Way
• Count the number of A, B, DR antigen mismatches – Why do so many zero-antigen mismatches fail?
–
Why are many mismatches successful?
• Determine unacceptable mismatches for highly sensitized patients – Why do see so many graft failures?
– Why are many patients never transplanted?
• Crossreactive antigen matching for platelet transfusions of refractory thrombocytopenic patients – Why are so many such transfusions unsuccessful?
– Why do some non-crossreactive mismatches work so well?
Important Consideration
Differentiate between Antigenicity of epitopes (reactivity with antibodies) and Immunogenicity of epitopes (induction of specific antibodies)
HLA Mismatch Immunogenicity
• Mismatched HLA antigens have different epitope “loads” • HLA epitopes have different degrees of immunogenicity A better understanding of HLA immunogenicity will permit a permissible mismatch strategy for non-sensitized transplant patients
KIDNEY ALLOGRAFT SURVIVAL AND HLA CLASS I MATCHING AT THE AMINO ACID TRIPLET LEVEL
Duquesnoy, RJ , Takemoto S, de Lange P, Doxiadis IIN, Schreuder GMT , Persijn, G and Claas FJH: Transplantation 75:884-889, 2003
Effect of HLA-A,B Triplet Mismatching on Graft Survival of Zero-HLA-DR Mismatched Kidneys in Eurotransplant
Mismatch Group: 0 AB Ag 0 Trp 1 Trp 2 Trp 3 Trp 4 Trp 5-9 Trp 10-19 Trp 20+ Trp Number of Tx: 3426 231 218 377 450 629 3448 6078 943 % Graft Survival after 1 year 2 years 89.3
87.1
3 years 5 years 83.7
76.8
89.2
85.1
82.3
75.6
86.7 85.7 89.1 88.4
84.3 82.1 86.7 84.1
80.7 78.3 82.4 81.1
76.4 72.9 75.4 75.9
Low epitope “load ” 85.1
81.5
77.4
69.9
85.4
80.6
76.7
69.8
84.3
80.0
76.1
68.5
HLAMatchmaker Predicts Antibody Formation to HLA Mismatches
Volume 77(8) 27 April 2004 pp 1236-1239
THE NUMBER OF AMINO ACID TRIPLET DIFFERENCES BETWEEN PATIENT AND DONOR IS PREDICTIVE FOR THE ANTIBODY REACTIVITY AGAINST MISMATCHED HUMAN LEUKOCYTE ANTIGENS
Dankers, Marlies K. A; Witvliet, Marian D; Roelen, Dave L; De Lange, Peter; Korfage, Nelleke; Persijn, Guido G.; Duquesnoy, René; Doxiadis, Ilias I. N; Claas, Frans H. J.
Department of Immunohematology and Blood Transfusion, Leiden University Medical Center
Volume 77(8) 27 April 2004 pp 1236-1239
THE NUMBER OF AMINO ACID TRIPLET DIFFERENCES BETWEEN PATIENT AND DONOR IS PREDICTIVE FOR THE ANTIBODY REACTIVITY AGAINST MISMATCHED HUMAN LEUKOCYTE ANTIGENS
Dankers, Marlies K. A; Witvliet, Marian D; Roelen, Dave L; De Lange, Peter; Korfage, Nelleke; Persijn, Guido G.; Duquesnoy, René; Doxiadis, Ilias I. N; Claas, Frans H. J.
Department of Immunohematology and Blood Transfusion, Leiden University Medical Center
Class II Epitope Load and Immunogenicity
Duquesnoy et al “ Retransplant Candidates Have Donor-Specific Antibodies that React with Structurally Defined HLA-DR,DQ,DP Epitopes” . Transplant immunology, 18:352-360, 2008
Anti-DRB1 and Anti-DRB3/4/5 Antibodies after Kidney Transplantation
Mismatch DRB1 DR51 (DRB*05) DR52 (DRB*03) DR53 (DRB*04) Donor-Specific Antibodies 23/96 (24%) 4/8 (50%) 6/13 (46%) 15/18 (83%) Nr of Mismatched Eplets 6.8 + 3.6* 9.8 + 2.5
11.2 + 1.0
13.2 + 2.3
*p< 0.01
Anti-DQ and anti-DRB1 Antibodies after Kidney Transplantation
Mismatch DRB1 (N=96) Donor-Specific Antibodies 24% Nr of Mismatched Eplets 6.8 + 3.6* DQB (N=62) DQA (N=74) 87% 64% 10.2 + 3.3
11.4 + 4.9
*p< 0.01
The Epitope “Load” Determines the Immunogenicity of a Mismatched HLA Antigen
How Important is DRB1 Matching in Kidney Transplantation?
HLA Class II Matching
• DRB1 is “standard” • DRB3/4/5 – Antibodies to DR51, DR52 and DR53 • DQB1 and DQA1 – Relevance of DQB matching in transplantation – Patients make antibodies to DQB and DQA epitopes • DPA1 and DPB1 – Relevance of DP matching – Anti-DP antibodies in transplantation
A DR Antigen Mismatch Represents an Extra Epitope Load DRB1+ DRB3/4/5+ DQB+DQA+DPB+DPA
Conventional Matching for a DR7,DR18 Patient
Patient DRB1*0302 DRB1*0701 Donor mmDRB DRB1*0101 DRB1*0301 DRB1*0302 DRB1*0401 DRB1*0701 DRB1*0801 DRB1*0901 DRB1*1001 DRB1*1101 DRB1*1201 DRB1*1301 DRB1*1401 DRB1*1501 DRB1*1601 self self Which is the “best” mismatch?
Allele Based Compatibility for DR7,DR18 Patient
All are common DR-DQ haplotypes Donor mmDRB DRB1*0101 DRB1*0301 DRB1*0302 DRB1*0401 DRB1*0701 DRB1*0801 DRB1*0901 DRB1*1001 DRB1*1101 DRB1*1201 DRB1*1301 DRB1*1401 DRB1*1501 DRB1*1601 DRW x DRB3*0101 DRB3*0101 DRB4*0101 DRB4*0101 x DRB4*0101 x DRB3*0202 DRB3*0202 DRB3*0101 DRB3*0202 DRB5*0101 DRB5*0202 DQB DQA DQB1*0501 DQA1*0101 DQB1*0201 DQA1*0501 DQB1*0402 DQA1*0401 DQB1*0301 DQA1*0302 DQB1*0201 DQA1*0201 DQB1*0402 DQA1*0401 DQB1*0303 DQA1*0302 DQB1*0501 DQA1*0101 DQB1*0301 DQA1*0501 DQB1*0301 DQA1*0501 DQB1*0603 DQA1*0103 DQB1*0502 DQA1*0104 DQB1*0602 DQA1*0102 DQB1*0502 DQA1*0102 Self Self DRB1*0302 DRB1*0701 DRB3*0101 DRB4*0101 DQB1*0201 DQB1*0402 DQA1*0301 DQA1*0401
Epitope-Based Compatibility for DR7,DR18 Patient All are common DR-DQ haplotypes
Donor mmDRB DRB1*0101 DRB1*0301 DRB1*0302 DRB1*0401 DRB1*0701 DRB1*0801 DRB1*0901 DRB1*1001 DRB1*1101 DRB1*1201 DRB1*1301 DRB1*1401 DRB1*1501 DRB1*1601 DRW x DRB3*0101 DRB3*0101 DRB4*0101 DRB4*0101 x DRB4*0101 x DRB3*0202 DRB3*0202 DRB3*0101 DRB3*0202 DRB5*0101 DRB5*0202 DQB DQA DQB1*0501 DQA1*0101 DQB1*0201 DQA1*0501 DQB1*0402 DQA1*0401 DQB1*0301 DQA1*0302 DQB1*0201 DQA1*0201 DQB1*0402 DQA1*0401 DQB1*0303 DQA1*0302 DQB1*0501 DQA1*0101 DQB1*0301 DQA1*0501 DQB1*0301 DQA1*0501 DQB1*0603 DQA1*0103 DQB1*0502 DQA1*0104 DQB1*0602 DQA1*0102 DQB1*0502 DQA1*0102 Self Self Patient DRB1*0302 DRB1*0701 DRB3*0101 DRB4*0101 DQB1*0201 DQB1*0402 DQA1*0301 DQA1*0401 TotEp 31 8 0 19 3 6 9 32 23 27 30 31 39 43 DRB 5 1 0 8 0 6 3 6 6 10 3 4 15 17 DQB 12 0 0 10 0 0 5 12 10 10 11 12 10 12 DQA 14 7 0 1 3 0 1 14 7 7 16 15 14 14
Are DR8, DR9 and DR17 more “desirable” mismatches?
Epitope-based HLA Class II Compatibility
• Epitope compatibility information requires high resolution DRB, DQ and DP types • It is possible to identify DR mismatches with relatively low epitope loads • Such mismatches may reduce anti-class II antibody responses and perhaps benefit transplant outcome ?
HLA Mismatch Immunogenicity
• Mismatched HLA antigens have different epitope loads
• HLA epitopes have different degrees of immunogenicity
Frequencies of antibodies to donor DQB eplet mismatches DQB Eplet Expressed on Nr of cases Antibody Frequency 79ED2 52PQ3 45GE5 55PPP 70GT 45EV 57PA 14GL5 77DR 45GV 74SV2 140A2 52PL3 84QL2 74EL2 14GM 26L 66DI 70RT 26YL3 DQ4 DQ1 DQ2 DQ3 DQB1*0602/3 DQ7 DQ2,8 DQ5 DQ2,5 DQ1,4,8,9 DQ4,5 DQ2,6 DQ3,4 DQ2,3,4 DQ3,6 DQ2,4,6,8 DQ2,8,9 DQB1*0602/3/4/9 DQ2,4,6s DQB1*0602 *0603 DQB1*03 *0601/4/9 6 18 24 14 19 16 24 14 15 18 25 15 12 12 27 14 11 15 15 13 100% 89% 88% 79% 68% 63% 50% 50% 30% 29% 29% 27% 27% 23% 14% 13% 11% 8% 0% 0%
Duquesnoy et al Transplant immunology, 18:352-360, 2008
Frequencies of antibodies to donor DQA eplet mismatches DQA Eplet Eplet on 160AE 41GR3 75SL4 47EK2 50EF11 48LF 56RR5 69L 60QF5 80IRS2 75ILR 56RB 47ERW 34HE 25YT 41ER 160AD DQA1*0501/5 DQA1*04 *05 *06 DQA1*05 DQA1*02 DQA1*01 DQA1*02 *03 DQA1*03 DQA1*02 *03 *05 DQA1*02 *03 *04 *05 *06 DQA1*02 *03 *04 *06 DQA1*02 *04 *06 DQA1*02 *04 *05 *06 DQA1*0101/4/5 DQA1*0101/4/5 *02 *03 DQA1*01/2/4 *04 *05 DQA1*0101/2/4/5 *02 *03 DQA1*01 *02 *0301 *04 *06 Nr of cases 17 31 10 15 24 12 16 15 23 21 16 21 14 10 17 22 20 Antibody Frequency 80% 74% 67% 56% 45% 43% 40% 35% 32% 30% 29% 23% 10% 7% 4% 0% 0%
Duquesnoy et al Transplant immunology, 18:352-360, 2008
Dominant Eplets Reacting with anti-HLA-DP Antibodies
Locus DPB DPB DPB Mismatched Eplet 84DEAV 55DE* 84DEAV and/or 55DE Eplet Carrying Alleles Antibody Frequency DPB1 *01 *03 *05 *06 *09 *10 *11 18/23 (78%) *13 *14 *16 *17 *19 *20 *21 *30 DPB1*0201 *03 *0402 *06 *09 *10 *14 *16 *17 *18 *20 15/19 (79%) Donor- Specific Antibody 5/6 (83%) 5/6 (83%) 27/34 (79%) 11/11 (100%) DPA DPA DPA 51RA,83A 51QA,83T Other Eplets DPA1 *02 *04 DPA1 *01 *03 18/56 (32%) 5/56/ (9%) 0/33 (0%) nd nd nd
*
55DE is similar to the 57DE eplet on DR11 All 55DE-reactive sera reacted also with DR11 Duquesnoy et al Transplant immunology, 18:352-360, 2008
The Antibody Response to an HLA Mismatch is Restricted to a Limited Number of Epitopes
SERUM ANALYSIS AFTER TRANSPLANT NEPHRECTOMY REVEALS RESTRICTED ANTIBODY SPECIFICITY PATTERNS AGAINST STRUCTURALLY DEFINED HLA CLASS I MISMATCHES
Adeyi et al. Transplant Immunology, 14: 53-62, 2005
Thirty Patients With Rejected Kidney Transplants Underwent Allograft Nephrectomy 100% PRA 80% 60% 40% 20% 0%
Before Tx Pre-AlloNx. Post-AlloNx
Adeyi et al. Transplant Immunology, 14: 53-62, 2005
Donor Epitopes: A3: 62Qe,66rNv,70aQs,76Vd,80gTl,144tKr,149aAh,151aHe,163dT A31: 56R,62Qe,66rNv,74iD,76Vd,80gTl,193Av B55: 131S B63: 45Ma,66rNm,70aSa,74Y,131S
Adeyi et al. Transplant Immunology, 14: 53-62, 2005
Donor Epitopes (serum-reactive epitopes are shown in underlined bold font) A3: 62Qe,66rNv,
70aQs
,
76Vd
,80gTl,144tKr,149aAh,
151aHe
,
163dT
A31:
56R
,62Qe,66rNv,
74iD
,
76Vd
,80gTl,193Av B55: 131S B63:
45Ma
,66rNm,
70aSa
,74Y,131S
Adeyi et al. Transplant Immunology, 14: 53-62, 2005
Donor Epitopes (serum-reactive epitopes are shown in underlined bold font) A3: 62Qe,66rNv,
70aQs
,
76Vd
,80gTl,144tKr,149aAh,
151aHe
,
163dT
A31:
56R
,62Qe,66rNv,
74iD
,
76Vd
,80gTl,193Av B55: 131S B63:
45Ma
,66rNm,
70aSa
,74Y,131S Unacceptable epitopes identify unacceptable antigens e.g. 45Ma is present on B13, B46, B57,B62, B63. B75, B76, B76
Adeyi et al. Transplant Immunology, 14: 53-62, 2005
Case 3 Patient HLA-A1,A30; B8,B18; DR17, Donor HLA-A1,A30; B8,B13; DR7,DR17 Graft Survival: 5 months; Graft Nephr.: 12 months 100% 80% 60% 40% 20% -6 -4 -2 0 2 4 6 8 10 12 14 16 18 20 22 24 Months after Nephrectomy 0% Donor Epitopes: B13: 41T,45Ma,
76En ,
80rTa,
82aLr
,
144tQl
,163E 82aLr is expressed by all Bw4-associated HLA-B antigens + A23, A24, A25 and A32 144tQl is unique for HLA-B13 After six months: the anti-76En and anti-82aLr antibodies have disappeared, Only anti-144tQl antibodies are detected
Adeyi et al. Transplant Immunology, 14: 53-62, 2005
Relative Immunogenicity of Eplets How often do mismatched eplets induce specific antibodies?
14th International HLA Workshop Project Analysis of donor-specific antibodies in patients whose rejected kidney transplants had been removed
Preliminary Results: Eplet Immunogenicity in 62 Cases
Eplet 116D 171H 113HD 116Y 207S 9S 9F 113HN 9H 113YN 45EE 183DA 193AV 163E 66IA 69AQS 245AS # Reactive 0/9 0/9 1/10 1/10 3/15 2/9 3/13 3/13 2/8 2/8 3/9 5/15 5/15 4/11 3/8 3/8 3/8 0% 0% 10% 10% 20% 22% 23% 23% 25% 25% 33% 33% 33% 36% 38% 38% 38% Eplet 211S 76VDT 253Q 275EL 113YR 113YH 41T 150AHE 45TE 62QR 163R 30DTL 69AQA 166DG 45KE 76ENI 62RR # Reactive 3/8 5/13 4/10 4/10 6/13 8/17 5/10 5/10 6/11 6/11 6/11 5/9 5/9 8/14 6/10 6/10 5/8 38% 38% 40% 40% 46% 47% 50% 50% 55% 55% 55% 56% 56% 57% 60% 60% 63% Eplet 76ANT 127K 66KA 81ALR 180E 150AHV 94TV 105SW 142MTT 145HAA 166ES 79RL 177DK 65RK 62EG 62GR 161D # Reactive 5/8 13/20 12/18 8/11 8/11 16/22 15/20 15/20 17/22 17/22 7/9 8/10 8/10 16/19 12/14 16/18 9/10
Duquesnoy, RJ and Claas FHJ: Progress Report of 14th International Histocompatibility Workshop Project on the Structural Basis of HLA Compatibility, Tissue Antigens, 69 (Suppl. 1): 1-5, 2007
63% 65% 67% 73% 73% 73% 75% 75% 77% 77% 78% 80% 80% 84% 86% 89% 90%
15
th
International Workshop Project on Epitope Immunogenicity
• Analyze post-allograft nephrectomy sera for antibodies against donor class I and class II epitopes • Serum screening with single alleles (Luminex) and by CDC • So far, 40 laboratories worldwide will contribute informative cases • For more information go to
http://www.15ihiws.org/project.php?n=13 To participate please contact Frans Claas or Rene Duquesnoy
HLA Mismatch Immunogenicity
• Mismatched HLA antigens have different epitope loads • HLA epitopes have different degrees of immunogenicity A better understanding of HLA immunogenicity will permit a permissible mismatch strategy for non-sensitized transplant patients
New Website
http://www.HLAMatchmaker.net
• Introductory articles and tutorials • HLAMatchmaker related publications – From UPMC – From other institutions • Free downloads of all programs – Class I and class II matching – Antibody analysis – Other programs • Contacts and communications