Transcript HLAMATCHMAKER: A MOLECULARLY BASED DONOR …

ASHI’s New World Symphony
A Master Piece With Five Movements
First Movement: Perfecting The Match
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?
New Developments in HLA
• Complete typing for all HLA loci
• Better methods for HLA antibody
identification
• HLA structure and polymorphisms
• HLA and transplant immunity
HLA Effect on Transplantation
•
Humoral Immunity
– Complement-dependent antibodies
– Complement-independent antibodies
•
Cellular Immunity
– Direct Allorecognition
• Cytotoxic CD8 T-cells
• Effector CD4 T-cells
• Regulatory T-cells
– Indirect Allorecognition
• Effector CD4 T-cells
– NK cells (KIR)
•
HLA-restricted Immune Responses
– Anti-viral immunity
• Cytotoxic CD8 T-cells
– Recurrent autoimmune disease
• Effector CD4 T-cells
•
Graft-versus-Host Reactivity
– Mediated by donor T-cells
Antibody Responses to HLA
• Class I HLA antigens
– HLA-A and HLA-B
– HLA-C
– MICA
• Class II HLA antigens
–
–
–
–
DRB1
DRB3, 4, 5
DQB and DQA
DPB and DPA
HLA Antigens Have Multiple Epitopes
• “Private” epitopes and “public” epitopes shared
between cross-reacting antigens (CREGs)
• Epitopes defined by polymorphic amino acid
residues
• HLAMatchmaker
– Original version (2002): “triplets”, i.e. linear threeresidue sequences
– New version (2006): “eplets”, i.e. patches of residues
within a 3 Angstrom radius of a polymorphic residue
on the molecular surface.
Duquesnoy: “A Structurally Based Approach to Determine HLA Compatibility at
the Humoral Immune Level”, Human immunology, 67: 847-862, 2006
Amino Acid Polymorphisms On
The Molecular Surface Are
Responsible for Epitopes That
Induce Specific Alloantibodies
How can we visualize the polymorphic residues?
Locations of Polymorphic Residues on HLA-A, B, C Molecules
From: Kostyu et al. Human Immunology 57, 1-18, 1997
Space Fill Model of HLA Class I Molecule
Top view
Side view
a1
peptide
a1
a2
a2
b2M
a3
Visualization of polymorphic residues with three-dimensional
structural modeling of HLA molecules with Cn3D program
downloadable from the NCBI website
www.ncbi.nlm.nih.gov
Polymorphic Residues on Class I Antigens
HLA-A2
HLA-B27
HLA-Cw3
Topography of Polymorphic Residues on
HLA-DR and HLA-DQ Molecules
HLA-DQ
HLA-DR
a1
a1
b1
b1
a2
b2
a2
b2
HLAMatchmaker Concept
The HLA type of the antibody producer
determines what structural components of an
immunizing HLA antigen can be “seen” as non-self
Structural Basis of a HLA-B51 Mismatch
Polymorphic
Residues on B51
Structural Basis of a HLA-B51 Mismatch
Polymorphic
Residues on B51
“Seen” by
A2,A68;
B27,B44
Structural Basis of a HLA-B51 Mismatch
Polymorphic
Residues on B51
“Seen” by
A2,A68;
B27,B44
“Seen” by
A2,A68;
B35,B44
Structural Basis of a HLA-B51 Mismatch
Polymorphic
Residues on B51
“Seen” by
A2,A68;
B27,B44
“Seen” by
A2,A68;
B35,B44
“Seen” by
A2,A24;
B7,B8
Matching at the Epitope Level
Provides a Better Assessment
of HLA Compatibility than
Matching at the Antigen Level
HLAMatchmaker: Original Version
Donor HLA-A,B mismatches are defined by
linear sequences of amino acid residues
(triplets) on alloantibody-accessible sites
(i.e. a-helices and b-turns) of HLA molecules
Human Immunol. 63: 339-352, 2002 + 63: 353-363, 2002
Histocompatibility Determination by
HLAMatchmaker
• The HLA-A,B,C type of the patient represents
six strings of self-triplets that cannot induce
specific alloantibodies
• Each donor HLA antigen represents a string
of triplets
• Compatibility is assessed by lining up donor
triplet strings with patient triplet strings to
determine differences between donor and
patient (Microsoft Excel program)
Human Immunol. 63: 339-352, 2002 + 63: 353-363, 2002
Notation System for Triplets and Eplets
The number indicates the amino acid sequence position
Polymorphic residues are listed with the single amino
acid letter code (monomorphic residues are not listed)
Examples: 9F 12SV …. 66RNV 76ES 90A …etc.
Potential Donor has
a HLA-B8 Mismatch
For which patient is this antigen
a better mismatch?
Patient 1: HLA-A2,A30; B18,B27; Cw2,Cw4
Patient 2: HLA-A2,A31; B42,B53; Cw2,Cw7
HLA-B8 Mismatch for
HLA-A2,A30; B18,B27; Cw2,Cw4
Patient HLA
A02
A*0201
A30
A*3001
B18
B*1801
B27
B*2705
Cw2 Cw*0202
Cw4 Cw*0401
Donor HLA
B8
B*0801
9
F
S
H
H
Y
S
12
sV
sV
sV
sV
aV
sV
14
R
R
R
R
R
W
17 41 45 56 62 66 70 74
gR A Me G Ge rKv aHs H
gS A Me R Qe rNv aQs D
GR A Te G Rn qIs tNt Y
gR A Ee G Re qIc aKa D
sR A Ge G Re qKy rQa D
gR A Ge G Re qKy rQa aD
D aM
R
gR
A Ee G Rn qIf
tNt
76
Vd
Vd
Es
Ed
Vn
Vn
80
gTl
gTl
rNl
rTl
rNl
rNl
D Es
rNl
Mismatched triplets are in underlined bold white font
HLA-B8 Mismatch for HLA-A2,A30; B18,B27; Cw2,Cw4
82
A02 lRg
A30 lRg
B18 lRg
B27 lLr
Cw2 lRg
Cw4 lRg
Donor HLA
B8 lRg
163
A02 T
A30 T
B18 T
B27 E*
Cw2 E*
Cw4 T
Donor HLA
B8
T
90 105 107 127 131 138 142
A
S W K
R
T T
A
S
G N
R
T I
A
P
G N
S
T I
A
P
G N
S
T I
A
P
G N
R
T I
D
P
G N
R
T I
144 147 149 151 156 158
tKh W aAh aHv L
A
tQr W aAr aRw L
A
tQr W aAr aRv L
A
tQr W aAr aRv L
A
tQr W aAr aRe W
A
tQr W aAr aRe R
A
A
P
G N
R
T I tQr W aAr
166 171 177 180 184 186 193 199 207 246
Ew Y Et Q A
K Av A
S
A
Ew Y Et Q P
K Pi A
G
A
Ew H Et Q P
K Pi A
G
A
Ew Y Et Q P
K Pi A
G
A
Ew Y Et Q Eh K Pv A
G
A
Ew Y Et Q Eh K Pv A
G
A
Ew
Y
Dt
E
P
K
Pi
A
G
A
aRv D
248 253
V
Q
V
E
V
E
V
E
V
E
V
E
V
A
E
HLA-B8 has six mismatched triplets
HLA-B8 Mismatch for
HLA-A2,A31; B42,B53; Cw2,Cw7
Patient HLA
9 12 14 17 41 45 56 62
66
70
74 76
A2
A*0201
F sV
R gR
A Me G Ge rKv aHs
A31
A*3101
T sV
R gR
A Me R Qe rNv aHs iD Vd gTL
B42
B*4201
Y sV
R gR
A Ee G Rn
B53
B*5301 Y aM R gR
Cw2
Cw*0202 Y aV R sR
Cw7
Cw*0701 D aV R gR
Donor HLA
B8
B*0801
D aM R gR
qIy aQa
H
80
Es
rNl
A Te G Rn qIf tNt Y En
A Ge G Re qKy rQa D Vn
A Ge G Re qNy rQa aD Vs
rIa
rKl
rNl
A Ee G Rn
rNl
qIf
tNt
D
Vd gTL
D
Es
82
90
105
107
127
131
138
142
144
147
149
151
156 158
A2
lRg
A
S
W
K
R
T
T
tKh
W
aAh aHv
L
A
A31
lRg
A
G
N
R
T
I
tQr
W
aAr aRv
L
A
B42
lRg
A
S
P
G
N
R
M
I
tQr
W
aAr aRv
D
A
B53
Cw2
Cw7
aLr
lRg
lRg
A
A
D
P
P
P
G
G
G
N
N
N
S
R
R
M
T
T
I
I
I
tQr
tQr
tQr
W
W
L
aAr aRv
aAr aRe
aAr aRa
L
W
L
A
A
A
B8
lRg
A
P
G
N
R
M
I
tQr
W
aAr aRv
D
A
163
166
171
177
180
184
186
193
199
207
246
248
253
A2
T
Ew
Y
Et
Q
dA
K
Av
A
S
A
V
Q
A31
T
Ew
Y
Et
Q
dP
K
Av
A
S
S
V
Q
B42
T
Ew
Y
Dt
E
dP
K
Pi
A
G
A
V
E
B53
L
Ew
Y
Et
Q
dP
K
Pv
A
G
A
V
E
Cw2
E
Ew
Y
Et
Q
eH
K
Pv
A
G
A
V
E
Cw7
T
Ew
Y
Et
Q
eP
K
Pl
A
G
A
V
Eq
B8
T
Ew
Y
Dt
E
dP
K
Pi
A
G
A
V
E
B8 is a zero-triplet mismatch for A2,A31;B42,B53;Cw2,Cw7
Examples of HLA Antigens with 0-2 Triplet Mismatches
Nr of
Mismatched
Triplets
Zero
Patient 1:
HLA-A2,A30;
B18,B27;
Cw2,Cw4
Patient 2:
HLA-A2,A31;
B42,B53;
Cw2,Cw7
A69, B64, B65
A32, A74, B8, B35,
B54, B55, B56, B59,
Cw3, Cw6
One
A68, B37, B39, B73, A69, B51, B67, B70,
B74, Cw6
B71, B72, B76, B78
Two
B61, B70, B71, B72, A33, A68, B7, B38,
B39, B46, B52, B58,
Cw1, Cw3, Cw6
B64, B65, B75, B77,
B81, B82, Cw1,
Cw8
Is HLAMatchmaker
Clinically Useful?
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
Five-Year Graft Survivals of Zero-HLA-DR Mismatched Primary
Kidney Transplants with 0-4 HLA-A,B Antigen Mismatches
UNOS Database
0 AB Ag
100
1 ABAg
2 AB Ag
3 AB Ag
90
4 AB Ag
80
P<0.001
70
60
0
1
2
3
4
5
Effect of HLA-A,B Triplet Matching on Five-Year Graft Survivals of
Zero-HLA-DR Mismatched Kidney Transplants in the UNOS Database
UNOS Database
0 ABDR(N=10609)
100
0-2T (N=251)
3-10T (N=4060)
>10T (16959)
90
>0DR
p<0.001
80
70
60
0
1
2
3
4
5
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
Number of Recipients: 3426
231
1 trp 2 trp 3 trp 4 trp 5-9 trp 10-19 trp 20-29 trp
218 377 450 629 3448
6078
943
% Graft Survival after
1 year
2 years
3 years
5 years
86.7
84.3
80.7
76.4
89.3
87.1
83.7
76.8
89.2
85.1
82.3
75.6
85.7
82.1
78.3
72.9
89.1
86.7
82.4
75.4
88.4
84.1
81.1
75.9
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
The Epitope “Load” Determines
the Immunogenicity of a
Mismatched HLA Antigen
HLA Matching at the Triplet Level
• Reduces antibody formation
• Improves kidney transplant survival
• Increases the availability of suitably
matched donors
The Highly Sensitized
Transplant Candidate
“Match” or “Treat”?
The Highly Sensitized
Transplant Candidate
“Match” or “Treat and Match”?
Transplant Strategy For Highly
Sensitized Candidates
•
•
•
•
•
Sensitization history
Laboratory assessment of sensitized state
Serum antibody reactivity and specificity
Identification of acceptable mismatches
Assess transplantability of patient (what is the
chance of finding a suitably matched donor?)
HLAMatchmaker for the
Highly Sensitized Patient
Example
Patient : HLA-A3,11;B18,62 (PRA=96%)
HLAMatchmaker Results:
Mismatched for
Zero Triplets
B75
One Triplet
B72 B76
Two Triplets
A30 B35 B46 B50 B54 B64 B70 B71 B78
Lymphocytotoxicity Screen
Patient : HLA-A3,11;B18,62
(PRA=96%)
1. 48/50 panel cells gave consistently positive reactions
with patient’s serum
2. Two negative cells: A3,A26;B62,- and A11,-;B18,51
3. A26 and B51 have unshared triplets but none of them
are apparently recognized by patient’s antibodies
9
12
14
17
A03 A*0301
F
sV
R
A11 A*1101
Y
sV
B18 B*1801
H
B62 B*1501
41
45
56
62
74
76
gR
A Me
G
Qe rNv aQs
D
Vd gTL
R
gR
A Me
G
Qe rNv aQs
D
Vd gTL
sV
R
gR
A
Te
G
Rn qIs tNt
Y
Es
rNl
Y
aM
R
gR
A Ma
G
Re qIs tNt
Y
Es
rNl
A26 A*2601
Y
sV
R
gR
A Me
G
Rn rNv aHs D
B51 B*5101
Y
aM
R
gR
A
G
Rn qIf tNt
82
90 105 107 127 131 138 142 144 147 149 151 156
Te
66
70
Y
80
An gTL
En rIa
A03 A*0301 lRg
A
S
G
N
R
T
I
tKr
W
aAh aHe
L
A11 A*1101 lRg
D
P
G
N
R
T
I
tKr
W
aAh aHa
Q
B18 B*1801 lRg
A
P
G
N
S
M
I
tQr
W
aAr aRv
L
B62 B*1501 lRg
A
P
G
N
S
M
I
tQr
W
aAr aRe W
A26 A*2601 lRg
D
P
G
N
R
T
I
tQr
W
tAh aHe W
B51 B*5101 aLr A
P
G
N
S
M
I
tQr
W
aAr aRe
L
158 163 166 171 177 180 184 186 193 199 207 246 248 253
A03 A*0301
A
dT Ew
Y
Et
Q
dP
K
Pi
A
G
A
V
E
A11 A*1101
A
R
Ew
Y
Et
Q
dP
K
Pi
A
G
A
V
E
B18 B*1801
A
T
Ew
H
Et
Q
dP
K
Pi
A
G
A
V
E
B62 B*1501
A
L
Ew
Y
Et
Q
dP
K
Pi
A
G
A
V
E
A26 A*2601
A
R
Ew
Y
Et
Q
A
S
S
V
Q
B51 B*5101
A
L
Ew
H
Et
dA K Av
Q dP K Pv
A
G
A
V
E
Patient 3: HLA-A3,11;B18,62 (Contd)
4. Eight mismatched triplets on A26 and five mismatched
triplets on B51 are not recognized by patient’s
antibodies; they are acceptable triplet mismatches
5. Enter A26 and B51 as “negative antigens” in
HLAMatchmaker
6. The program will identify additional HLA antigens with
triplets that either matched or acceptable
Patient 3: HLA-A3,11;B18,62 (PRA=96%)
HLAMatchmaker Results after Serum Analysis
Neg Ag
Neg Ag
Zero-trp mm
Mismatched for
Zero/Acc Triplets: A25 A26 A32 A66 A74 B35 B51 B52 B53 B75 B77
One Triplet
A29 A34 A43 B59 B72 B76
Two Triplets
A30 A31 A33 B46 B49 B50 B54 B63 B64 B70 B71
After A26 and B51 had been entered as negative antigens, HLAMatchmaker
identified eight HLA antigens as acceptable/zero triplet mismatches
Serum Screening May Identify
Acceptable Antigen Mismatches
and Increase the Probability of
Finding a Suitable Donor for a
Highly Sensitized Patient
Chances of Finding a Zero or Acceptable HLA-A,B
Mismatch for a High PRA Patient (30 patients)
Caucasoid Donors
10.000%
Zero/Acceptable Triplet mm
Zero/Acceptable Ag mm
1.000%
0.100%
0.010%
29
27
25
23
21
19
17
15
13
11
9
7
5
3
1
0.001%
Patient Number
Eurotransplant Patients, Leiden, The Netherlands
Eurotransplant’s Acceptable Mismatch
Program Now Includes HLAMatchmaker
Validation of HLAMatchmaker by
direct cross-matches
HLA-A mismatch
Expected Observed
negative negative
0 triplet 18
18
1 triplet 25
25
HLA-B mismatch
Expected
negative
0 triplet
54
1 triplet 133
Observed
negative
54
131
AM are Acceptable Mismatches
5-85% and >85% PRA are
conventional ET-KAS cases
Claas et al.: The Acceptable Mismatch Program as a Fast Tool to Transplant Highly
Sensitized Patients Awaiting a Post-Mortal Kidney: Short Waiting Time and Excellent Graft
Outcome. Transplantation 78:190-193, 2004
Shortcomings of Current
HLAMatchmaker Algorithm
• Limited to linear three-residue
sequences (i.e. triplets)
• Does not always consider
conformational influences of nearby
polymorphic residues
• Triplets do not identify all corresponding
serologically defined determinants
New Eplet Version of
HLAMatchmaker
•
Duquesnoy, RJ: “A Structurally Based Approach to Determine HLA Compatibility
at the Humoral Immune Level”, Human immunology, 67: 847-862, 2006
•
Duquesnoy, RJ and Askar, M: “HLAMatchmaker: A Molecularly Based Algorithm
for Histocompatibility Determination. V. Eplet Matching for HLA-DR, HLA-DQ
and HLA-DP“, Human Immunology, 68: 12-25, 2007
New HLAMatchmaker Version
• Based on molecular structures of protein-antibodies
complexes and identification of contact residues with the
specificity-determining CDRs such as CDR-H3
• Considers all surface-exposed polymorphic residues
and residues within a 3.0-3.5 Angstrom radius
• These polymorphic residues clusters will be referred to
as “eplets” rather than triplets because eplets may
include nonlinear sequences with more than three
residues
• Requirement: All well-defined serological determinants
should have corresponding eplets
Polymorphic Residue Positions in 3.0 Angstrom Patches of HLA Class I Antigens
Sequence Class I
Position Locus
1
C
6
C
9
ABC
12
BC
14
C
17
A
30
B
32
B
35
C
41
B
44
A
45
B
56
A
62
A
63
ABC
65
ABC
66
ABC
69
B
70
ABC
71
B
73
AC
76
A
77
ABC
79
A
80
ABC
82
AB
83
AB
90
ABC
94
BC
Molecular
Location
side
side
side
underside
side
side
underside
underside
side
side
side
side
side
top
top
top
top
top
top
side
top
top
top
top
top
top
top
side
underside
Surface
Exposure
++
+
-/+
+
++
++
+
+
+
++
++
+
+
+
-/+
++
++
++
+
+
++
++
+
++
++
++
++
++
-/+
Polymorphic Positions within
3.0 Angstroms
1
6
113
9
11 12
14 17
14 17
30
32
35
41
43 44
44 45
56
62 63
62 63 66
65 66 67
63 65 66 67 70 P2
69 70
66 69 70 71
70 71
69 73 74 77
76 77 80
73 76 77 P8 P9
79 80
76 78 80 81
81 82 83
82 83
90
94
Sequence Class I
Position Locus
103
BC
105
A
107
A
109
A
113
BC
114
ABC
116
ABC
127
A
131
B
138
C
142
A
143
B
144
A
145
AB
147
BC
149
A
150
A
151
A
152
ABC
158
AB
161
A
163
A
166
A
167
AB
173
C
177
ABC
178
B
180
B
184
AC
186
A
193
AC
194
ABC
199
B
207
A
211
AC
219
AC
245
A
248
C
253
AC
261
C
267
C
268
C
270
C
275
C
276
A
Molecular
Location
side
side
side
side
underside
underside
underside
side
side
side
top
top
side
top
top
top
top
top
top
top
top
top
top
top
side
side
side
side
side
side
bottom
side
bottom
side
side
bottom
side
side
bottom
side
side
side
side
side
side
Surface
Exposure
-/+
++
++
+
+
-/+
-/+
++
++
++
++
+
+
++
+
++
++
++
+
++
++
++
++
++
++
++
++
++
++
++
++
++
+
+
-/+
++
+
++
++
+
++
++
++
++
++
Polymorphic Positions within
3.0 Angstroms
103
105
107
109
6
113 114
113 114
116
127
131
138
142 143
143 145 147
143 144 145
144 145 149
143 147
145 149 150
149 150 151
150 151 152
151 152
158
161
163 167
166 167
163 166 167
173
177 178
177 178
180
184
186
193 194
193 194
199
207
211
219
245
248
253
261
267 268
267 268
270
275
276
HLA-ABC Eplets
• Provisional repertoire: 203 eplets, 125 on top, 76 on
side and 32 in less accessible positions
• Many eplets are equivalent to triplets
• Excellent correlations between eplets and
serologically defined private and public determinants
Class II Eplet Version of
HLAMatchmaker
Duquesnoy, RJ and Askar, M: “HLAMatchmaker: A Molecularly Based Algorithm
for Histocompatibility Determination. V. Eplet Matching for HLA-DR, HLA-DQ
and HLA-DP“, Human Immunology, 68: 12-25, 2007
New Class II Version Considers
DR, DQ and DP
• DRB1 and DRB3/4/5
– Patients can make antibodies to DR51, DR52 and DR53
• DQA1 and DQB1
– Patients can make antibodies to DQB and DQA
epitopes
Topography of Polymorphic Residues on
HLA-DR and HLA-DQ Molecules
HLA-DQ
HLA-DR
a1
a1
b1
b1
a2
b2
a2
b2
Class II Version Considers DR, DQ and DP
• DRB1 and DRB3/4/5
– Patients can make antibodies to DR51, DR52 and DR53
• DQA1 and DQB1
– Patients can make antibodies to DQB and DQA
epitopes
• DPA1 and DPB1
– Clinical relevance of anti-DP antibodies in
transplantation
Sequence Molecular Surface
Position Location Exposure 3.0 Angstrom Patches
3.0 Angstrom
Patches on
DRB1,3,4,5
Polymorphic
positions are
underlined
4
6
12
14
16
18
25
26
31
32
33
34
40
41
44
47
48
51
57
58
59
60
67
70
Side
Side
Underside
Underside
Underside
Side
Side
Side
Underside
Underside
Underside
Side
Side
Side
Side
Side
Side
Side
Top
Top
Top
Top
Top
Top
++
+
+
+
+
+
++
+
+
+
+
++
+
+
+
+
++
+
+
+
++
++
+
+
3
5
11
13
15
17
16
25
10
31
8
33
28
40
41
28
47
37
56
54
58
59
66
67
4
6
12
14
16
18
24
26
29
32
32
34
39
41
43
46
48
50
57
57
59
60
67
69
5
7 A15 A17
13 29
15 16 27 29
17 18 25
19 23
25 26 43
27 42
30 31 32
33 35
33 34
35 A83
40 41
42 43 44 45
44 45
47 48 62
49
51 52
58 61 A76 P13
58 59 62
60
61 63
68 71
70 71 73 P11
Sequence Molecular Surface
Position Location Exposure 3.0 Angstrom Patches
DRB1,3,4,5
Contd
71
73
74
76
77
81
85
86
96
98
104
105
108
112
120
133
135
140
142
149
180
181
183
187
189
Top
Top
Top
Top
Top
Top
Top
Side
Side
Side
Side
Side
Side
Bottom
Side
Bottom
Bottom
Side
Side
Side
Side
Side
Side
Side
Bottom
+
+
+
++
++
+
+
+
++
++
+
++
++
++
+
++
++
++
++
++
+
++
+
++
++
70
69
70
73
73
80
84
82
95
97
103
104
107
108
98
132
134
139
138
148
96
180
182
186
188
71
72
71
75
76
81
85
85
96
98
104
105
108
111
119
133
135
140
141
149
177
181
183
187
189
72
73
72
76
77
82
86
86
97
99
105
106
109
112
120
134
136
141
142
150
179
182
184
188
190
73
74
73
77
78
85
76
74
P6
P4
87 90
180
120
107 114
107
113
121
143
180 181
77
79
P8
P9
Polymorphic Residues in 3 Angstrom DRB Patches
4
6
11
13
16
18
16
25
10
31
8
33
28
40
41
28
47
51
57
57
58
59
67
12 13
14 16
18 25
25
26
30
32
32
34
40
41
44
47
48
26
31 32
33
33 34
41
44
48
58
58 59
59 60
60
71
67
70
73
70
73
81
85
96
98
104
108
112
120
133
135
140
142
149
180
183
187
189
70
71
74
71
76
85
86
180
120
105
181
71 73
73
76 77
73 74
77
DQA
2
18
21
25 26
40 41
44 45
34 47
48 51
50 51
48 50
51 52
51 52
51 53
54 55
53 55
59 61
64
66
69
75 76
75 76
75 79
107
129 130
138 139
153
156
160 161
163
175
48
51 52 53
53
53 54
54 55
56
56
79
80
DQB
3
13
23
26
30
45
45
46
49
52
52
55
56
66
66
70
67
74
76
84
84
86
85
86
116
125
130
135
140
167
182
14
37
46
46
47
53
55
56
57
67
67
71
70
75
77
85
85
87
86
89
126
168
38
47
56
57
71
71
86 89
89 90
90
DPA
18
28
50 51
72 73
83
111
127
160
DPB
8
11
28
33
35
43
56
64
64
65
43
76
84
84
85
86
91
96
171
173
36
70
57
65
65 69
69 70
69 70
85
85 86
86 87
87
Polymorphic Residues in 3 Angstrom
DQ and DP Patches
Determination of the Polymorphic
Residue Composition of Each Patch
• HLA Patch Generator: a Microsoft Excel
Macro developed by Grzegorz Dudek
(Medigen Molecular Diagnostics, Warszawa ,
Poland)
Eplet Numbers on HLA Class II Molecules
Locus # Eplets
DRB
149
DQB
74
DQA
58
DPB
45
DPA
19
Serologically Defined DRB Antigens and Corresponding Unique Eplets
DRB1 Antigen
DR1
DR2
DR3
DR4
DR7
DR8
DR9
DR10
DR11
DR12
DR14
Unique Eplets
12LKF
14REH
73GRDN
14HEH
14YKH
25YRF
12DKF
12VKF
59EEY
25YRL
60EH
31QCIY
133L
142M
34HQ
96YL
98EN 180LT
25HQF
31QLFY 76GDT
73ALDT
70FRRA
32VHN
40EYD 74RRAA
31YHFH
47EFR
183A
51R
44NL
104AR
48YQ
108T
DRB3,4,5 Antigen
DRB3 (DR52)
DRB3*01 (DR52a)
DRB3*02 (DR52b)
DRB4 (DR53)
DRB5 (DR51)
DRB5*01(DR51a)
DRB5*02(DR51b)
98QS
12RKS
31LHFH
25HWN
12DKY
31QDIY
6C
81YV 96QM 180MM 187Q
Serologically Defined DQB Antigens and Corresponding Unique Eplets
DQ Antigen
DQ1
DQ5
DQ6
DQ2
DQ3
DQ7
DQ8
DQ4
Unique Eplets
52PQ
30HYV 71VGA 87AY 116I
87AF 125GQ
30SIV 45GE 52LL 67DIK
55PPP
45EV
56PPA
55PRL
DQA Alleles and Unique Corresponding Eplets
Unique Eplets
DQA1*01
18F
48WF 56KGG 59PG 69A 76IM 129QS 175Q
DQA1*02
47EKL 55LR.
DQA1*03
25YS 47EQL 52FRR 75IVR
DQA1*04/06 69T
DQA1*05
76SL 107I
156L
163S 175K
DQA1*0602 138TR
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 DR Compatibility
Patient
DRB1*1101 DRB1*1301
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
Epitope-Based DR Compatibility
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
DQB1*0501
DQB1*0201
DQB1*0402
DQB1*0301
DQB1*0201
DQB1*0402
DQB1*0303
DQB1*0501
DQB1*0301
DQB1*0301
DQB1*0603
DQB1*0502
DQB1*0602
DQB1*0502
DQA
DQA1*0101
DQA1*0501
DQA1*0401
DQA1*0302
DQA1*0201
DQA1*0401
DQA1*0302
DQA1*0101
DQA1*0501 Self
DQA1*0501
DQA1*0103 Self
DQA1*0104
DQA1*0102
DQA1*0102
Patient
DRB1*1101 DRB1*1301 DRB3*0202
DQB1*0603 DQB1*0301 DQA1*0103
TotEp
21
14
16
39
47
15
39
25
0
9
5
19
18
30
DRB
7
5
5
26
29
4
25
11
0
9
5
4
16
18
DQB
10
9
7
0
9
7
1
10
0
0
0
10
0
10
x
DQA1*0501
DQA
4
0
4
13
9
4
13
4
0
0
0
5
2
2
Epitope-Based DR/DQ Compatibility for DR7,DR18 Patient
Patient
DRB1*0302 DRB1*0701 DRB3*0101
DQB1*0202 DQB1*0402 DQA1*0301
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
DQB1*0501
DQB1*0201
DQB1*0402
DQB1*0301
DQB1*0201
DQB1*0402
DQB1*0303
DQB1*0501
DQB1*0301
DQB1*0301
DQB1*0603
DQB1*0502
DQB1*0602
DQB1*0502
DQA
DQA1*0101
DQA1*0501
DQA1*0401
DQA1*0302
DQA1*0201
DQA1*0401
DQA1*0302
DQA1*0101
DQA1*0501
DQA1*0501
DQA1*0103
DQA1*0104
DQA1*0102
DQA1*0102
Self
Self
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
DRB4*0101
DQA1*0401
DQA
14
7
0
1
3
0
1
14
7
7
16
15
14
14
Epitope-Based DR Compatibility for DR7,DR18 Patient
Patient
DRB1*0302 DRB1*0701 DRB3*0101
DQB1*0202 DQB1*0402 DQA1*0301
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
DQB1*0501
DQB1*0201
DQB1*0402
DQB1*0301
DQB1*0201
DQB1*0402
DQB1*0303
DQB1*0501
DQB1*0301
DQB1*0301
DQB1*0603
DQB1*0502
DQB1*0602
DQB1*0502
DQA
DQA1*0101
DQA1*0501
DQA1*0401
DQA1*0302
DQA1*0201
DQA1*0401
DQA1*0302
DQA1*0101
DQA1*0501
DQA1*0501
DQA1*0103
DQA1*0104
DQA1*0102
DQA1*0102
Self
Self
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
DRB4*0101
DQA1*0401
DQB
12
0
0
10
0
0
5
12
10
10
11
12
10
12
Are DR8, DR9 and DR17 more “desirable” mismatches?
DQA
14
7
0
1
3
0
1
14
7
7
16
15
14
14
Epitope-based HLA Class II Compatibility
• Epitope compatibility information requires highresolution 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
To be Continued:
First Movement Practicale,
Fine-tuning the Instrument