HLA: matching and donor
selection
Dr Bronwen Shaw
Consultant in haematopoietic cell transplantation
Royal Marsden Hospital
Anthony Nolan Trust
Overview
• Why does HLA matter in transplantation?
• HLA
•
•
•
•
Where is it found?
What does it do?
Tissue typing?
What does the nomenclature mean and how do we
classify ‘matching’ and ‘mismatching’?
• Polymorphism
• Some examples
HCT Component Data
Survival for CML Unrelated HCT: JMDP Tally Effect
What is HLA? What is it for?
• Human Leukocyte Antigen
• Discovered: in mice (1937), humans (1954)
• Function: to present peptides to T cells, thus
allowing elimination of foreign particles and
recognition of self (so in transplants this has to be
modulated)
4 Mb
HLA-A
3 Mb
HLA-C
HLA-B
2 Mb
1 Mb
HLA-DR
HLA-DQ
HLA-DP
0 Mb
The HLA Family
A1
A2
Cw7 Cw5
B8
B44
The HLA Family
A1
A2
Cw7 Cw5
B8
B44
A3
A26
Cw7 Cw8
B7
B14
The HLA Family
A1
A2
Cw7 Cw5
B8
B44
A1
A3
Cw7
Cw7
B8
B7
A3
A26
Cw7 Cw8
B7
B14
The HLA Family
A1
A2
A3
Cw7 Cw5
B8
B44
A1
A3
Cw7
Cw7
B8
B7
A26
Cw7 Cw8
B7
A1
A26
Cw7 Cw8
B8
B14
B14
The HLA Family
A1
A2
A3
Cw7 Cw5
B8
B44
A1
A3
Cw7
Cw7
B8
B7
A26
Cw7 Cw8
B7
A1
A26
Cw7 Cw8
B8
B14
A2
A3
Cw5 Cw7
B44
B7
B14
The HLA Family
A1
A2
A3
Cw7 Cw5
B8
B44
A1
A3
Cw7
Cw7
B8
B7
A26
Cw7 Cw8
B7
A1
A26
Cw7 Cw8
B8
B14
A2
A3
A2
A26
Cw5 Cw7
Cw5 Cw8
B44
B44
B7
B14
B14
The HLA Family
A1
A2
A3
Cw7 Cw5
B8
B44
A1
A3
Cw7
Cw7
B8
B7
A26
Cw7 Cw8
B7
A1
A26
Cw7 Cw8
B8
B14
A2
A3
A2
A26
Cw5 Cw7
Cw5 Cw8
B44
B44
B7
B14
B14
A1
A3
Cw7
Cw7
B8
B7
The HLA Family
A1
A2
A3
Cw7 Cw5
B8
B44
A1
A3
Cw7
Cw7
B8
B7
A26
Cw7 Cw8
B7
A1
A26
Cw7 Cw8
B8
B14
A2
A3
A2
A26
Cw5 Cw7
Cw5 Cw8
B44
B44
B7
B14
B14
A1
A3
Cw7
Cw7
B8
B7
The HLA Family
A1
A2
A3
Cw7 Cw5
B8
B44
A1
A3
Cw7
Cw7
B8
B7
A26
Cw7 Cw8
B7
A1
A26
Cw7 Cw8
B8
B14
A2
A3
A2
A26
Cw5 Cw7
Cw5 Cw8
B44
B44
B7
B14
B14
A1
A3
Cw7
Cw7
B8
B7
The HLA Family
A1
A2
A3
Cw7 Cw5
B8
B44
A1
A3
Cw7
Cw7
B8
B7
A26
Cw7 Cw8
B7
A1
A26
Cw7 Cw8
B8
B14
A2
A3
A2
A26
Cw5 Cw7
Cw5 Cw8
B44
B44
B7
B14
B14
The HLA Family
A1
A2
A3
Cw7 Cw5
B8
Cw7 Cw8
B44
A1
A3
Cw7
Cw7
B8
B7
A26
B7
A1
A26
Cw7 Cw8
B8
B14
A2
A3
A2
A26
Cw5 Cw7
Cw5 Cw8
B44
B44
B7
B14
B14
A1
A3
Cw7
Cw7
B8
B7
MHC Sequencing Consortium, 1999
• One of the most gene-dense regions of the genome
• 224 genes identified within the 3.6 Mb
• 40% of the genes have functions within the immune
response
• HLA is the most polymorphic region in the human genome
HN RP A 1 { }
DR8
DR1
6
DR51
5 6
DR53
487
Classical Class II
(0.8 Mb)
DR52
Extended Class II
(0.3 Mb)
NO T CH 4
G18
PB X2
RA G E
LPAA T
G16
NG 3
P P T2
NG 5
NG 7
T NX B
CR E B L 1
P 4 5 0 - C 2 1B
C4 B
ST K19
DO M 3 L
S K I2W
RD
BF
{ }
C4AQO, C4B1
C4A, C4B,C4B, C4B
C4A,C4B, C4B
C4A,C4B
Cen
Classical Class I (1.8 Mb)
{ }
K IA A O O 5 5 { }
{ }
M IC B
H C G IX - 1 {  }
3 .8 -1 .1
P 5 -1
M IC A
HL A -X
H C G IX - 2 {  }
N O B 1 { }
N O B 3 { }
N O B 2 { }
Z F4 7 L
h s 6 M 1 - 2 2 { }
h s 6 M 1 - 2 3 { }
Z NF 5 7 -L
M OG
G A B B R1
S M T 3 B { }
h s 6 M1 -1 2
r P L 1 3 { }
F A T1 0
h s 6 M 1 - 1 3 { }
h s 6 M 1 - 1 4 { }
M a s P O { }
r P S 1 7 { }
T RE { }
M a s -G P C R - L
M a s -G P C R - L {  }
h s 6 M1 -1 6
F A T 1 0 { }
h s 6 M1 -1 7
h s 6 M1 -1 8
h s 6 M1 -2 0
h s 6 M 1 - 1 9 { }
h s 6 M1 -2 7
h s 6 M1 -2 1
DD X 6 -L { }
h s 6 M1 -2 8
ZNFs
h s 6 M 1 - 2 4 { }
h s 6 M 1 - 2 5 { }
h s 6 M 1 - 7 { }
h s 6 M1 -6
h s 6 M 1 - 5 { }
h s 6 M1 -3
h s 6 M 1 - 2 { }
h s 6 M1 -4
h s 6 M1 -1
h s S A R A { }
h s 6 M1 -1 5
h s 6 M 1 - 2 6 { }
Z NF9 1 L
h s 6 M 1 - 8 { }
C Y T O K E R A T IN
RF P
N O P 5 6- L
B B C 1 { }
Z F P 3 8 -L
GPX 5
Z NF 9 1 -L
h s 6 M 1 - 9 { }
G P X 5 { }
S R E -Z B P
Z N F 2 0 - L { }
Z NF 4 7 -L
Z N L 43 - L { }
P R D5 1
T O B 4 { }
ORs
P 5 - 8 { }
P R D 5 1 { }
Z NF1 6 5
S R E - Z B F { }
HL A -B
H C G IV - 1 {  }
DH FR P { }
H L A - 1 7 { }
H C G II- 1 {  }
h s 6 M 1 - 3 1 { }
h s 6 M 1 - 2 9 { }
h s 6 M 1 - 3 5 { }
R P L3 - h o m {  }
h s 6 M 1 - 3 0 { }
K IA A 0 0 36
Extended Class I (4 Mb)
h s 6 M1 -3 2
h s 6 M 1 - 3 4 { }
Tel
HL A -C
H C G IV - 2 {  }
h s 6 M1 -1 0
h s 6 M 1 - 3 3 { }
H2 A F N
HS B F N-L
H3 F J
H1 F 5
H3 F F
H4 F K
H2 B F D
H 2 B F ID
H2 A F I
H2 A F E
H2 A F D
H 3 F 1K
H 4 F 2i i i
H 4 F 2i v
HB FE
HISTONEs
N O B 5 { }
H C G IX - 3 {  }
H C G II- 2 {  }
P O U 5 F1
N O B 4 { }
T CF1 9
HC R
S P R1
SE EK1
CD S N
ST G
P52
DD R1
F LO T IL L IN
IE R 3
H2 B F C
H2 A F C
r P L 2 4 { }
H 4 F F P { }
P O M 1 2 1 -L
PR SS16
Z NF1 8 4
H B 15 { }
r P L 2 { }
h s 6 V 1 - 5 { }
h s 6 V 1 - 4 { }
h s 6 V 1 - 3 { }
h s 6 V 1 - 1 { }
C 1 1 H 1 .2 - L {  }
h s 6 V 1 - 2 { }
r P L 1 0 { }
H2 A F A
H2 B F Q ii
h n R N P A 1 -L { }
H2 B F A iii
H 4 F 2i i
H 3 { }
H2 A F A L ii
Z N F - 1 8 4 -L { }
S M A 3 -L
M P - L { }
G A P DH-L { }
B T N2 A 1
BT N
b G L U - L { }
B T N3 A 2
B T N3 A 1
B T N2 A 2
B T N2 A 3
B T N3 A 3
H3 n 1
H 1 { }
H2 B F G
H2 A F G
H2 B F A ii
H4 F J
R P S 1 0 { }
H3 F D
H2 A F O L ii
H1 F 3
H 2 A { }
H4 n 2
H3 F I
H4 n 1
H3 F H
H2 B F J
H2 B _ n o ve l
H 4 _ n o v el
H2 B F K
H 2 A F K { }
H2 B _ n o v e l
H _ n o v e l { }
H2 B _ n o ve l
H2 A _ n o ve l
H 1 _ n o v el
S L C1 7 A 2
H 4 _ n o v el
H 1 _ n o v el
H4 F 2
H1 F 1
Ro Re t
H2 A F O L i
H3 F 2
H3 F C
H2 B F Q i
H1 F 2
VR1 s
K IA A 0 1 70
T UB B
DB P 2
R P L7 A
F B19
P R O A - h om {  }
A B C5 0
CA T5 6
HS R1
HL A -E
T C4
H C G II- 3 {  }
M IC C {  }
H C G II- 4 {  }
H C G II- 5 {  }
G T 25 7
H L A - 3 0 { }
CA T7 5 X
Z NF1 7 3
H L A - 9 2 { }
Z NFB 7
RF B 3 0
HC G I
HC G V
HT E X 4
H C G V III- 1
H C G V II
H L A - 5 9 { }
H C G IV - 3 {  }
P 5 - 2 { }
3 .8 -1 .2 {  }
H C G IX - 4
M IC D {  }
H C G II- 6 {  }
H L A - 8 0 { }
H C G IV - 4 {  }
HL A -A
P 5 - 3 { }
P 5 - 4 { }
H C G IV - 5 {  }
HL A -7 0
H L A - 2 1 { }
P 5 - 5 { }
H C G IV - 6
P 5 - 6 { }
3 .8 -1 .3 {  }
H C G II- 7
H L A - 1 6 { }
P 5 - 7 { }
H L A - 5 4 { }
P 5 - 9 { }
H C G IV - 7 {  }
P 5 - 1 0 { }
3 .8 -1 .4 {  }
M IC F {  }
HL A -G
H C G V III- 2 {  }
H C G IV - 8 {  }
P 5 - 1 1 { }
H C G II- 8 {  }
P 5 - 1 2 { }
R P L7 B
H L A - 9 0 { }
H C G IV - 9 {  }
P 5 - 1 3 { }
H L A - 7 5 { }
H 4 F 2i
HF E
H2 B F A 1
H1 F T
H2 A F A L 1
S L C1 7 A 1
S L C1 7 A 3
S L C1 7 A 2
BTNs
HS E T
A P T -2
r P L 1 2 -L { }
r P L 3 5 A - L { }
DA X X
B IN G 3 { }
T APB P
B IN G 1
RG L 2
HK E 2
B IN G 4
RP S 1 8
B 3 G A LT 4
B IN G 5 { }
HS A CM 2 L
T A T - S F 1 - L { }
Class III (0.7 Mb)
R IN G 1
Z N F - L { }
R IN G 2
C2
G10
NG 3 6
NG 2 2
G9a
NE U
G8
HS P A 1 B
HS P A 1 A
HS P A 1 L
s n RN P
G7c
V a l-T R S
NG 2 3
M S H5
C L IC 1
DD A H
G6c
G6b
G6d
G6e
G6f
B A T5
G5c
G5b
CS K 2 B
B A T4
G4
ApoM
B A T3
B A T2
H C G IV - 1 0 {  }
P 5 - 1 4 { }
3 .8 -1 .5 {  }
H C G IX - 5 {  }
HL A -F
M IC E { }
H C G IV - 1 1 {  }
HISTONEs
R IN G 5
RX RB
CO L 1 1 A 2
H L A - D P A 3 { }
H L A - D P B 2 { }
C O L 1 1 A 2 { }
HL A -D P B 1
H L A - D P A 2 { }
HL A -D P A 1
R P L3 2 - L { }
HL A -D O A
R IN G 3
HL A -D MA
HL A -D MB
R IN G 1 3 {  }
H L A - Z 1 { }
R IN G 1 4 {  }
IP P 2 {  }
LMP2
R IN G 8 {  }
T AP1
R IN G 9 {  }
LMP7
T AP2
HL A -D O B
H L A - D Q B 2 { }
H L A - D Q A 2 { }
H L A - D Q B 3 { }
C O X 3 -L {  }
G L N - tR N A {  }
HL A -D Q B 1
HL A -D Q A 1
1 C7
A IF 1
L TB
LST 1
T NFa
L TA
P 5 - 1 5 { }
IE
HL A -D RB 1
HL A -D RB 3
HL A -D RB 2 {  }
B A T1
N F K B IL1
AT P6G
R
HL A -D RA
HL A -D RB 9 { }
B T L -II
T SBP
B 3 0 .2 - L { }
C
A
R
R
s
ORs
Map of the human MHC on chromosome 6
HLA Class I
- found on all nucleated cells
Polymorphism located in exons 2 & 3
exon
1
2
3
4
5
6 7
8
protein
domain
L
a
a
a
TM
CYT
3'UT
Class I HLA-A
HLA Class II
- restricted to cells of the
immune system
Polymorphism located in exon 2
exon
1
2
3
4
5
a-chain gene
protein
domain
L
exon
1
a
2
a
3
4 5
TM/CYT
3'UT
6
b-chain gene
protein
domain
L
b
b TM CYT 3'UT
Class II HLA-DR
HLA and Tissue Typing
Cell
HLA and Tissue Typing
Cell
Serology-low resolution
e.g. A2
HLA and Tissue Typing
Medium resolution ‘string’
e.g. A*0201/0205/0209/0240
SSP, SSOP
Cell
High resolution (definitive)
e.g. A*02010101
SBT
HLA Nomenclature
• Gene names
HLA-A or HLA-DRB1
• Antigen names
A2 or DR1
• Allele names
A*020101 or DRB1*01010101
HLA Allele Nomenclature
HLA - A * 24 02 01 01
HLA - A * 24 02 01 02 L
Locus
Asterisk Allele family
(serological
where
possible)
Amino
Non-coding
Intron,
N = null
acid
(silent)
3’ or 5’
L = low
difference polymorphism polymorphism S = Sec.
A = Abr.
Level of resolution
Antigen matched
Low level of resolution
A*02
Medium level (string)
A*0201/0205/0209/0240
Type is ONE OF these four: Antigen matched
BUT do not know if allele matched
High level
A*020101
Allele matched
Ambiguity
•
Using medium level resolution typing it is possible to exclude some but
not all alleles from a group, hence the National Marrow Donor Program
(NMDP) codes.
B*1501 or B*1502
=
B*15AB
B*1501/1502/1505/1515/1521/1545/1556/1570=
•
B*15FGR
This is important for donor selection i.e. you may be able to tell if a donor is
definitely MISMATCHED but not matched
Numbers of HLA antigens and alleles 1968 - 2004
120 New HLA Class I alleles per year
50 New HLA Class II alleles per year
Number of HLA Alleles June 2004/2006/2007
(http://www.ebi.ac.uk/imgt/hla/stats.html)
HLA- A
HLA- B
HLA-C
325 (24)
489/617
592 (49)
830/960
175 (9)
210/335
DRB
DQA1
DQB1
DPA1
DPB1
458 (20)
545
(DRB1 463/542)
28
57 (7)
22
103
34
78/87
23
125/127
MICA
TAP
56
11
Figures in parenthesis indicate the number of serologically defined antigens at each locus.
Linkage Disequilibrium/ Haplotypes
• LD: Alleles occur together with a greater frequency than
would be expected by chance
– B/C strong, DR/DQ strong, A less strong, DP weak
– e.g. B*0801 - 99% will be Cw*0701
– But B*1801 either Cw*0501, *0701
– e.g DRB1*1501 - will be DQB1*0602
– But DRB1*0401 either Cw*0301, *0302
• Haplotype: A group of genes inherited together
– e.g. A*0101,B*0801,Cw*0701,DRB1*0301,DQB1*0201
Examples for search coordinators
and data managers
Example 1: VUD search
• Patient:
– A*0101, B*0801, DRB1*0301
• Finding a donor:
– Common haplotype - therefore likely
– Usually in strong LD - therefore ‘predictable’
– Even at low resolution (A1, B8, DR3) good
chance of being matched
Example 1: Data entry
• Patient:
– A*0101, B*0801, DRB1*0301
• Donor:
– A1, B8, DRB1 03: Antigenic match for A, B,
DRB1
• No data can be entered for C, DQB1, DPB1
• No data can be entered for allele level matching
– A*0101, B*0801, DRB1*0301: Allelic and
antigenic match for A, B, DRB1
• No data can be entered for C, DQB1, DPB1
Example 2: VUD search
• Patient:
– A*0201, B*1801, DRB1*0401
• Finding a donor:
– Less strong LD - therefore ‘ NOT predictable’
i.e. equal chance of C being *0701, *0501,
*1203; DQB1 50/50 chance of *0301, *0302
– At low resolution (A2, B18, DRB4) unable to
predict this will match
Example 2: VUD search (cont)
• Donor
A*02
Example 2: VUD search (cont)
• A*02
Example 2: VUD search (cont)
• A*02
Example 2: VUD search (cont)
• A*02
Example 2: VUD search (cont)
• A*02
Example 2: VUD search (cont)
• A*02
Example 2: VUD search (cont)
• A*02
Example 2: Data entry
• Patient:
– A*0201, B*1801, DRB1*0401
• Donor:
– A2, B18, DRB1 04: Antigenic match for A, B, DRB1
• No data can be entered for C, DQB1, DPB1
• No data can be entered for allele level matching
– A*0201, B*1801, DRB1*0401: Allelic and antigenic
match for A, B, DRB1
• No data can be entered for C, DQB1, DPB1
– A*0201/05, B*1801, DRB1*0401: Allelic and
antigenic match for B, DRB1
• Antigenic match for A BUT cannot say that this is an allelic
match
Example 2: Data entry
• Patient:
– A*0201, B*1801, DRB1*0401
• Donor:
– A2, B18, DRB1 04: Antigenic match for A, B, DRB1
• No data can be entered for C, DQB1, DPB1
• No data can be entered for allele level matching
– A*0201, B*1801, DRB1*0401: Allelic and antigenic
match for A, B, DRB1
• No data can be entered for C, DQB1, DPB1
– A*0201/05, B*1801, DRB1*0401: Allelic and
antigenic match for B, DRB1
• Antigenic match for HLA-A BUT cannot say that this is an
allelic match
Example 3: search and data
patient
donor
search
data
A*0201/9207/920 A*0201/9207/920 Probable match
9
9
(rare)
Select donor
Antigen match
(probable
allelic)
B*3503
B*4403
B*3503
B*4403
Match
Select donor
Antigenic and
allelic match
Cw*0401/24
Cw*1601
Cw*0401/20/24
Cw*1601
Probable match
(rare)
Select donor
Antigen match
(probable
allelic)
DRB1*0701,
DRB1*1501/18
DRB1*0701,
DRB1*1501/18
Probable match
(rare)
Select donor
Antigen match
(probable
allelic)
Match
Antigenic and
allelic match
Rare alleles: A*9207, *9209, Cw*0420, Cw*0424, DRB1*1518
DQB1*0202
DQB1*0202
Example 3: search and data
patient
donor
search
data
A*0201/9207/920 A*0201/9207/920 Probable match
9
9
(rare)
Select donor
Antigen match
(probable
allelic)
B*3503
B*4403
B*3503
B*4403
Match
Select donor
Antigenic and
allelic match
Cw*0401/24
Cw*1601
Cw*0401/20/24
Cw*1601
Probable match
(rare)
Select donor
Antigen match
(probable
allelic)
DRB1*0701,
DRB1*1501/18
DRB1*0701,
DRB1*1501/18
Probable match
(rare)
Select donor
Antigen match
(probable
allelic)
Match
Antigenic and
allelic match
Rare alleles: A*9207, *9209, Cw*0420, Cw*0424, DRB1*1518
DQB1*0202
DQB1*0202
Example 3: search and data
patient
donor
search
data
A*0201/9207/920 A*0201/9207/920 Probable match
9
9
(rare)
Select donor
Antigen match
(probable
allelic)
B*3503
B*4403
B*3503
B*4403
Match
Select donor
Antigenic and
allelic match
Cw*0401/24
Cw*1601
Cw*0401/20/24
Cw*1601
Probable match
(rare)
Select donor
Antigen match
(probable
allelic)
DRB1*0701,
DRB1*1501/18
DRB1*0701,
DRB1*1501/18
Probable match
(rare)
Select donor
Antigen match
(probable
allelic)
Match
Antigenic and
allelic match
Rare alleles: A*9207, *9209, Cw*0420, Cw*0424, DRB1*1518
DQB1*0202
DQB1*0202
Example 4: search and data
patient
donor
search
A*0101-03/06
A*0201-14/16-22/2431/33/36-41
A*0101-03/06/07
A*0201-14/16-22/2431/33/36-42/44/45
Probable match - but
not certain
Antigen match
Cannot report allelic status
B*0801/05/07/10
B*4402/05/11/14
B*0801/05/07/10/12
B*4402/05/11/14
Probable match - but
not certain
Antigen match
Cannot report allelic status
Cw*0701/05/06/08/14
Cw*0401/03/04/05/06
/07
Cw*0701/05/06/08/14
Cw*0501-04
Probable match (one
antigen)- but not
certain
Definite mismatch
Antigen match x1
Cannot report allelic status
Antigen and allelic mm x1
DRB1*1301/40
DRB1*1401
DRB1*1301
DRB1*1401
Probable match - but
not certain
Antigen match x1
Cannot report allelic status
Antigen and allelic mm x1
DQB1*0603/14
DQB1*050301
DQB1*0603/14
DQB1*050301
Probable match - but
not certain
Antigen match x1
Cannot report allelic status
Antigen and allelic mm x1
Rare alleles: ???????????
data
Useful websites
• Anthony Nolan Trust:
www.anthonynolan.org.uk
• NMDP allele codes:
http://bioinformatics.nmdp.org/HLA/Allele_Codes/Allele_Code_Lists/
index.html
• World Marrow Donor association:
www.worldmarrow.org
• Bone marrow donor worldwide:
www.bmdw.org
• HLA database:
http://www.ebi.ac.uk/imgt/hla/