HLA: matching and donor selection Dr Bronwen Shaw Consultant in haematopoietic cell transplantation Royal Marsden Hospital Anthony Nolan Trust
Jan 09, 2016
HLA: matching and donor selection
Dr Bronwen ShawConsultant 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
Survival for CML Unrelated HCT: JMDP Tally EffectHCT Component DataHCT Component Data
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
3 Mb
2 Mb
1 Mb
0 Mb
HLA-A
HLA-C
HLA-B
HLA-DR
HLA-DQ
HLA-DP
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
Cw7
B8
A1 A2
Cw7 Cw5
B8 B44
A3 A26
Cw7 Cw8
B7 B14
A3
Cw7
B7
The HLA Family
A1
Cw7
B8
A1 A2
Cw7 Cw5
B8 B44
A3 A26
Cw7 Cw8
B7 B14
A1 A26
Cw7 Cw8
B8 B14
A3
Cw7
B7
The HLA Family
A1
Cw7
B8
A1 A2
Cw7 Cw5
B8 B44
A3 A26
Cw7 Cw8
B7 B14
A1 A26
Cw7 Cw8
B8 B14
A2
Cw5
B44
A3
Cw7
B7
A3
Cw7
B7
The HLA Family
A1
Cw7
B8
A1 A2
Cw7 Cw5
B8B44A3 A26
Cw7 Cw8
B7 B14
A1 A26
Cw7 Cw8
B8 B14
A2
Cw5
B44
A3
Cw7
B7
A3
Cw7
B7
A2
Cw5
B44
A26
Cw8
B14
The HLA Family
A1
Cw7
B8
A1 A2
Cw7 Cw5
B8 B44
A3 A26
Cw7 Cw8
B7 B14
A1 A26
Cw7 Cw8
B8 B14
A2
Cw5
B44
A3
Cw7
B7
A3
Cw7
B7
A2
Cw5
B44
A26
Cw8
B14
A1
Cw7
B8
A3
Cw7
B7
The HLA Family
A1
Cw7
B8
A1 A2
Cw7 Cw5
B8 B44
A3 A26
Cw7 Cw8
B7 B14
A1 A26
Cw7 Cw8
B8 B14
A2
Cw5
B44
A3
Cw7
B7
A3
Cw7
B7
A2
Cw5
B44
A26
Cw8
B14
A1
Cw7
B8
A3
Cw7
B7
The HLA Family
A1
Cw7
B8
A1 A2
Cw7 Cw5
B8 B44
A3 A26
Cw7 Cw8
B7 B14
A1 A26
Cw7 Cw8
B8 B14
A2
Cw5
B44
A3
Cw7
B7
A3
Cw7
B7
A2
Cw5
B44
A26
Cw8
B14
A1
Cw7
B8
A3
Cw7
B7
The HLA Family
A1
Cw7
B8
A1 A2
Cw7 Cw5
B8 B44
A3 A26
Cw7 Cw8
B7 B14
A1 A26
Cw7 Cw8
B8 B14
A2
Cw5
B44
A3
Cw7
B7
A3
Cw7
B7
A2
Cw5
B44
A26
Cw8
B14
The HLA Family
A1
Cw7
B8
A1 A2
Cw7 Cw5
B8 B44
A3 A26
Cw7 Cw8
B7 B14
A1 A26
Cw7 Cw8
B8 B14
A2
Cw5
B44
A3
Cw7
B7
A3
Cw7
B7
A2
Cw5
B44
A26
Cw8
B14
A1
Cw7
B8
A3
Cw7
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
P5-15
{
}
HLA-
FHC
GIV-11
{
}
MICE {
}
HCGIX
-5 {
}
P5-14
{
}HC
GIV-1
0 {
}HL
A-75
{
}P5
-13 {
}
HCGIV
-9 {
}
HLA-
90 {
}
RPL7
B {
}HC
GII-8
{
}P5
-12 {
}
HCGIV
-8 {
}
P5-11
{
}HL
A-G
HCGV
III-2 {
}
MICF {
}
3.8-1
.4 {
}P5
-10 {
}
P5-9
{
}HL
A-54
{
}P5
-7 {
}
HLA-
16 {
}
HCGII
-73.8
-1.3 {
}
P5-6
{
}HC
GIV-6
P5-5
{
}HL
A-70
HLA-
21 {
}
HCGIV
-5 {
}
P5-4
{
}HL
A-A
P5-3
{
}HC
GIV-4
{
}HL
A-80
{
}HC
GII-6
{
}MIC
D {
}HC
GIX-4
3.8-1
.2 {
}P5
-2 {
}
HCGIV
-3 {
}
HLA-
59 {
}
HCGV
III-1HC
GVII
HTEX
4
3.8-1
.5 {
}
HCGIV
-7 {
}
HCGV
RFB3
0HC
GIZN
FB7
ZNF1
73
GT25
7HL
A-30
{
}HC
GII-5
{
}HC
GII-4
{
}MIC
C {
}HC
GII-3
{
}
HLA-
92 {
}
CAT7
5X
Tel
Cen
Classical Class I (1.8 Mb)
BTL-I
IHL
A-DR
AHL
A-DR
B3HL
A-DR
B2 {
}
HLA-
DRB1
HLA-
DQA1
HLA-
DQB1
GLN-
tRNA {
}
COX3
-L {
}
HLA-
DQB3
{
}HL
A-DQ
A2 {
}
HLA-
DQB2
{
}HL
A-DO
BTA
P2LM
P7TA
P1RIN
G9 {
}
LMP2
RING8
{
}IPP
2 {
}RIN
G14 {
}
HLA-
Z1 {
}
HLA-
DMB
RING1
3 {
}HL
A-DM
ARIN
G3HL
A-DO
AHL
A-DP
A1RP
L32-L
{
}HL
A-DP
B1HL
A-DP
A2 {
}
COL11
A2 {
}
HLA-
DPB2
{
}HL
A-DP
A3 {
}
COL11
A2RX
RBRIN
G5RIN
G2RIN
G1ZN
F-L {
}
TAT-S
F1-L
{
}HS
ACM2
LRP
S18
B3GA
LT4BIN
G5 {
}
HLA-
DRB9
{
}
Classical Class II (0.8 Mb)
Extended Class II (0.3 Mb)
BAT1
NFKB
IL1AT
P6G
LTA TNFa
LST1
1C7
AIF1
BAT2
BAT3
ApoM
G4 BAT4
CSK2
BG5
bG5
cBA
T5G6
fG6
eG6
dG6
cG6
bDD
AHCL
IC1
MSH5
NG23
G7c
Val-T
RSsn
RNP
HSPA
1LHS
PA1A
HSPA
1BG8 NE
UNG
22G9
aNG
36G1
0C2 BF RD SK
I2WDO
M3L
STK1
9C4
BP4
50-C
21B
TNXB
CREB
L1NG
7
LTB NG5
NG3
PPT2
LPAA
TG1
6
G18
NOTC
H4
RAGE
PBX2
Class III (0.7 Mb)
TC4
HSR1
HLA-
E
CAT5
6AB
C50
PROA
-hom {
}
DBP2
RPL7
A {
}KIA
A017
0TU
BBFL
OTILL
INIER
3DD
R1P5
2
TCF1
9PO
U5F1
NOB4
{
}HC
GII-2
{
}HC
GIX-3
{
}
HLA-
CHC
GIV-2
{
}KIA
AOO5
5 {
}RP
L3-ho
m {
}HC
GII-1
{
}HL
A-B
HCGIV
-1 {
}
DHFR
P {
}HL
A-17
{
}P5
-8 {
}
NOB2
{
}NO
B3 {
}
NOB1
{
}HC
GIX-2
{
}MIC
AHL
A-X {
}
P5-1
3.8-1
.1HC
GIX-1
{
}MIC
B
FB19
NOB5
{
}
B30.2
-L {
}
TSBP
HNRP
A1 {
}
BING4
HKE2
RGL2
TAPB
PBIN
G1DA
XXBIN
G3 {
}
rPL35
A-L {
}
rPL12
-L {
}
APT-2
DR8
6DR1
65
74 8DR53
DR51
C4A, C4B,C4B, C4BC4A,C4B, C4BC4A,C4B
DR52
C4AQO, C4B1
Extended Class I (4 Mb)
HFE
RFP
hs6M
1-8 {
}
hs6M
1-15
hs6M
1-26 {
}
hsSA
RA {
}
hs6M
1-1hs
6M1-4
hs6M
1-3hs
6M1-2
{
}hs
6M1-6
hs6M
1-5 {
}
hs6M
1-7 {
}
hs6M
1-25 {
}
hs6M
1-24 {
}
hs6M
1-23 {
}
hs6M
1-22 {
}
hs6M
1-21
hs6M
1-20
hs6M
1-19 {
}
hs6M
1-18
hs6M
1-17
hs6M
1-16
FAT1
0 {
}Ma
s-GP
CR-L
{
}Ma
s-GP
CR-L
rPS17
{
}Ma
sPO {
}
hs6M
1-14 {
}
FAT1
0hs
6M1-1
3 {
}rPL
13 {
}
hs6M
1-12
GABB
R1SM
T3B
{}
MOG
ZNF5
7-L
DDX6
-L {
}
HSET
H2AF
AL1
H2BF
A1H1
FTH4
F2i
H1F2
H3FC
H2BF
QiH2
AFOL
iH3
F2H4
F2H1
F1Ro
Ret
SLC1
7A2
H2BF
KH2
AFK {
}
H3FH
H2BF
JH3
FIH4
n1H4
n2H1
F3H2
A {
}H3
FDH2
AFOL
iiH2
BFAii
H4FJ
RPS1
0 {
}H2
BFG
H2AF
GH3
n1H1
{
}BT
N3A3
BTN2
A3BT
N3A1
BTN2
A2BT
N3A2
BTN2
A1bG
LU-L
{
}SM
A3-L
MP-L
{
}GA
PDH-
L {
}
H2AF
ALii
H2BF
AiiiH4
F2ii
H3 {
}
H2AF
AH2
BFQii
hs6V
1-2 {
}
rPL1
0 {
}
ZNF-
184-L
{
}
hs6V
1-1 {
}
POM1
21-L
PRSS
16ZN
F184
hnRN
PA1-L
{
}
HB15
{
}rPL
2 {
}
C11H
1.2-L
{
}
rPL24
{
}H4
FFP {
}
H2BF
CH2
AFC
H3F1
KH4
F2iii
H4F2
iv
H4_n
ovel
H1_n
ovel
H2B_
nove
lH2
A_no
vel
H1_n
ovel
H2B_
nove
lH_
nove
l {
}H2
B_no
vel
H4_n
ovel
HBFE
H2AF
EH2
AFD
H2BF
DH2
BFID
H2AF
IH1
F5H3
FFH4
FKH3
FJH2
AFN
HSBF
N-L
hs6M
1-10
KIAA0
036
hs6M
1-30 {
}
hs6M
1-31 {
}
hs6M
1-29 {
}
hs6M
1-35 {
}
ZNF1
65SR
E-ZB
F {
}PR
D51 {
}
ZF47
LPR
D51
TOB4
{
}ZN
L43-L
{
}ZN
F47-L
ZNF2
0-L {
}
ZNF9
1-LSR
E-ZB
Phs
6M1-9
{
}GP
X5 {
}
GPX5
ZFP3
8-LNO
P56-L
BBC1
{
}
CYTO
KERA
TINZN
F91L
hs6M
1-28
hs6M
1-27
TRE
{}
STG
CDSN
SEEK
1SP
R1HC
R
hs6M
1-33 {
}
hs6M
1-32
hs6M
1-34 {
}
BTN
HISTONEs HISTONEsBTNs ZNFsORs ORs
hs6V
1-3 {
}
hs6V
1-4 {
}
hs6V
1-5 {
}
SLC1
7A1
SLC1
7A3
SLC1
7A2
VR1s
Map of the human MHC on chromosome 6
1
L
exon
proteindomain
5
TM
8
3'UT
6 7
CYT
2 3 4
Polymorphism located in exons 2 & 3
HLA Class I- found on all nucleated cells
Class I HLA-A
1
L
exon
proteindomain
5
3'UT
-chain gene
2 3 4
TM/CYT
1
L
exon
proteindomain
6
3'UT
-chain gene
3 42 5
TM CYT
Polymorphism located in exon 2
HLA Class II - restricted to cells of the immune system
Class II HLA-DR
HLA and Tissue Typing
Cell
HLA and Tissue Typing
CellSerology-low resolution
e.g. A2
HLA and Tissue Typing
Cell
Medium resolution ‘string’e.g. A*0201/0205/0209/0240
SSP, SSOP
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
Locus Asterisk Allele family(serological
where possible)
Aminoacid
difference
Non-coding(silent)
polymorphism
Intron, 3’ or 5’
polymorphism
N = nullL = low
S = Sec.A = Abr.
HLA - A * 24 02 01 02 L
Low level of resolution A*02
Medium level (string) A*0201/0205/0209/0240
High level A*020101
Level of resolution
Antigen matched
Allele matched
Type is ONE OF these four: Antigen matched BUT do not know if allele matched
• 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
Ambiguity
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) 592 (49) 175 (9)489/617 830/960 210/335
DRB DQA1 DQB1 DPA1 DPB1
458 (20) 28 57 (7) 22 103545 (DRB1 463/542) 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 dataA*0201/9207/9209 A*0201/9207/9209 Probable match (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)
DQB1*0202
DQB1*0602
DQB1*0202
DQB1*0602
Match
Select donor
Antigenic and allelic match
Rare alleles: A*9207, *9209, Cw*0420, Cw*0424, DRB1*1518
Example 3: search and data
patient donor search dataA*0201/9207/9209 A*0201/9207/9209 Probable match (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)
DQB1*0202
DQB1*0602
DQB1*0202
DQB1*0602
Match
Select donor
Antigenic and allelic match
Rare alleles: A*9207, *9209, Cw*0420, Cw*0424, DRB1*1518
Example 3: search and data
patient donor search dataA*0201/9207/9209 A*0201/9207/9209 Probable match (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)
DQB1*0202
DQB1*0602
DQB1*0202
DQB1*0602
Match
Select donor
Antigenic and allelic match
Rare alleles: A*9207, *9209, Cw*0420, Cw*0424, DRB1*1518
Example 4: search and data
patient donor search dataA*0101-03/06
A*0201-14/16-22/24-31/33/36-41
A*0101-03/06/07
A*0201-14/16-22/24-31/33/36-42/44/45
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 certainDefinite 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: ???????????
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/