Tissue Typing in Aotearoa

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Tissue Typing in Aotearoa
Paul Dunn Clinical Scientist ASHI Director New
Zealand Blood Service Auckland paul.dunn_at_nzblood.c
o.nz ASHI American Society for
Histocompatibility Immunogenetics
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• ASHI Accreditation (03-09-NZ-01-1)
• Technologies/Methods Accredited
• Serology Typing class I and II
• DNA Typing class I/II low/medium
  microarrays, SSP
• DNA Typing class I/II high SSP
• Antibody screening/identification class I CDC,
  microarrays
• Antibody screening/identification class II
  microarrays
• Crossmatching T cells and B cells CDC, Flow
• Areas of Accreditation
• HSC/BM Transplantation related donor
• HSC/BM Transplantation unrelated donor
• Solid Organ Transplantation living donor
• Solid Organ Transplantation deceased donor
• Histocompatibility Testing for other
  non-clinical purposes
• Transfusion support

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Histotrac (SystemLink)
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HLA TESTING (and technology) 1/11/07 31/10/08
MagNA PURE DNA Extraction Robot
5,800 DNA extractions 8,000 HLA typing
(DNA) 2,500 PRA 5,100 HLA antibody screens 1,500
HLA antibody IDs 266 Flow Cytometry
crossmatches 305 CDC crossmatches
Hamilton Starlet
Luminex 1. SSO DNA Typing 2. HLA antibody
screening
Flow Cytometer patient/ live donor crossmatches
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TRANSPLANTATION STATISTICS 2005-06
MagNA PURE DNA Extraction Robot
Luminex - SSO DNA Typing (One Lambda)
- HLA antibody screening
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Test Menu 1 New Zealand Tissue Typing Laboratory

• BMT (or HSCT) patient HLA-A, -B, -DRB(x2)
  Luminex SSO typing (low/medium
• resolution)
• - potential related donor HLA-A, -B.
• Matching sib HLA-DRB and take 2nd sample
  (confirm types, patient
• donor).
• - No match, high resolution HLA-DRB type
  patient (i.e. DRB11101),
• search for unrelated donor (Bone Marrow
  Donors Worldwide 12M
• potential URDs 330,000 cord units).
• - High resolution type patient and donor
  (HLA-A,-B,-C, -DQB1,-DRB1)
• SSP/SBT.
• Aim match patient and unrelated donor, allele
  level 5 HLA loci.
• Chance of finding a matching urd 75-90

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Test Menu 2 New Zealand Tissue Typing Laboratory
Solid Organ patient new patient HLA-A, -B,
-DRB (x2). ABO 2x. Luminex SSO typing (low
resolution) - Luminex HLA antibody screen and
CDC PRA every 3 months. Joins waiting list
for deceased donor organ. - living donor
(kidney). HLA and ABO tests as above. Also, CDC
and flow crossmatch (patient serum donor
cells). Negative crossmatch, then repeat
crossmatch lt 1 week before transplant. -
deceased donor. HLA type (serology and SSP), ABO
. Matching run. Crossmatch (CDC) vs top 15
ABO-compatible patients.
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• RENAL TRANSPLANT PATIENT
• HLA Typing
• Relation HLA-A
  HLA-B HLA-DRB1
  HLA-DQB1 HLA-DPB1
• Patient TC 02, 02 15, 3502
  11, 12 02,
  0302 0201, 0402
• Antibody
• Test
  Result PRA No
  Tested Specificity
• B cell PRA
  POS 23 30
  IgM A3
• Luminex class I PRA
  POS 18
  A25, A26, A34, A66 (IgG), A3 (IgM)
• Luminex class II single antigen id POS
  
  DQ5, DQ6, DP1, DP3, DP5, DP9, DP10, DP11,
  DP13, DP14, DP17, DP19
• Donor Typing
• Donor Relation HLA-A
  HLA-B HLA-DRB1
  HLA-DQB1 HLA-DPB1
• ED brother 02, 11 35, 44 04,
  15 03, 06 0401
• DR brother 02, 03 15, 57 12,
  13 03, 06 0401, 0402
• BJG unknown 02, 11 35, 44
  01, 04 03, 0501 0401,
  1001
• LAW unknown 11, - 08,
  35 03, 0407 02, 03 0301,
  0401

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Test Menu 3 New Zealand Tissue Typing
Laboratory

• Bone Marrow Donors and Platelet Donors
  HLA-A,-B,-DRB1
• BM donors recruited mainly from
  Maori/Polynesian communities
• Random apheresis donors ABDR typed for B cell
  PRA
• Disease Association HLA type
• Transfusion support HLA antibody screen,
  crossmatch (TRALI). HLA and
• Human Platelet Antigen (HPA) antibody screen,
  HPA genotype (NAIT).
• Platelet provision for refractory patients.
• Pharmacogenomics
• HLA-B5701 and hypersensitivity to abacavir
• HLA-B1502 and hypersensitivity to carbamezapine
• Luminex SSO typing (low resolution)

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Test Menu 3 New Zealand Tissue Typing
Laboratory

• Bone Marrow Donors and Platelet Donors
  HLA-A,-B,-DRB1
• Disease Association HLA type
• Transfusion support HLA antibody screen,
  crossmatch (TRALI). HLA and
• Human Platelet Antigen (HPA) antibody screen,
  HPA genotype (NAIT).
• Platelet provision for refractory patients.
• Pharmacogenomics
• HLA-B5701 and hypersensitivity to abacavir
• HLA-B1502 and hypersensitivity to carbamezapine
• Luminex SSO typing (low resolution)

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HLA and disease associations
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Test Menu 3 New Zealand Tissue Typing
Laboratory

• Bone Marrow Donors and Platelet Donors
  HLA-A,-B,-DRB1
• Disease Association HLA type
• Transfusion support
• TRALI (transfusion-related acute lung
  injury) HLA antibody screen,
• crossmatch, neutrophil antibody screen.
• Dunn Dinesh (NZ Med J 2008 121(1276)42-7)
  17 cases (06/04-06/07) 29/67 donors HLA Ab
  86 female. 15 cases HLA Ab against patient
  TT.
• NAIT (neonatal alloimmune thrombocytopenia)
  HLA and
• Human Platelet Antigen (HPA) antibody screen,
  HPA genotype.
• Platelet provision for refractory patients
  (HLA matched/compatible).
• Pharmacogenomics HLA-B5701 and
  hypersensitivity to abacavir
• Luminex SSO typing (low resolution)

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Patient Refractory to Platelets HLA
Typing Patient VM HLA-A24, 33 B07, 14
DRB10102, 15 Antibody Test
Result
PRA Specificity Luminex HLA
class I id Positive
84 Multispecific Luminex
single antigen class I Positive
A11, A29, B13,
B2708, B27, B35,

B37, B38, B39, B4005,
B41, B44,

B45, B47, B48, B49, B50,
B51,

B52, B53, B56, B57, B58, B59,


B60, B61, B62, B63, B67, B71,


B72, B75, B77 Result select platelet donors
with acceptable mismatches. Select local ones.
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Test Menu 3 New Zealand Tissue Typing
Laboratory

• Bone Marrow Donors and Platelet Donors
  HLA-A,-B,-DRB1
• Disease Association HLA type
• Transfusion support HLA antibody screen,
  crossmatch (TRALI). HLA and
• Human Platelet Antigen (HPA) antibody screen,
  HPA genotype (NAIT).
• Platelet provision for refractory patients.
• Pharmacogenomics HLA-B5701 and
  hypersensitivity to abacavir
• Testing since November 2006 100 samples, 5
  B57 pos (5), 1 B5701
• HLA-B1502 carbamezapine (Stevens-Johnson
  syndrome)
• HLA-B5801 allopurinol
• Luminex SSO typing (low resolution)

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EQAs

• UCLA Cell Exchange PBLs class I serology. DNA
  extract (Luminex).
• UCLA Cell Exchange DNA extracts class I high
  res (SSP/SBT).
• UCLA sera antibody specificity id (CDC,
  Luminex).
• International DNA exchange (UCLA) class II
  high res (SSP).
• ASEATTA PBLs class I and II serology (local
  and commercial sera), DNA extract.
• ASEATTA DNA Exchange class I and II low res
  (SSP) (Laboratory sendout).
• ASEATTA sera antibody specificity id (CDC,
  Luminex).
• ASEATTA Flow Crossmatch (Laboratory sendout).
• ASHI Crossmatch/antibody id (CDC, Flow/CDC,
  Luminex).
• NEQAS Scheme 1B.

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• Research Projects (ethics approval)
• Donor Specific HLA antibody and chronic rejection
  in heart and lung transplant patients.
• B27 subtyping
• cDNA Project
• Maori DNA Typing
• Fetal Genotyping
• Molecular blood grouping
• Microarrays

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New Zealand Tissue Typing Laboratory - staff