HLA Selected Platelets

Dr Helen North PhD

Clinical Scientist

Department of Histocompatibility & Immunogenetics

NHS Blood and Transplant Colindale

Helen North
 Poor increment (<10 x 109/L or CCI <7.5 ) after at
least two consecutive transfusions of random
donor platelets
Platelet count taken from one hour to 24hours
post transfusion.
2011 NHSBT recognised count could be taken
after 10 minutes

Helen North
 Platelet Refractoriness
Causes:

immunological and non-immunological

Helen North
 Old/poorly stored platelets, small dose
Splenomegaly, hepatomegaly
DIC (infection, septicaemia, malignancy)
Infection (CMV)
Fever
Antibiotics, amphotericin B, ambisome,
vancomycin, ciprofloxacin,

Helen North
 Immunological causes
Preformed antibodies in recipient to antigens on
platelets:
HLA - class I specific antibodies (HLA-A,-B)
Most common cause of immune refractoriness
Rarely HLA -C
HPA antibodies (HPA-1a, -2b etc)
Incompatibility for HPA is uncommon
ABO - antibodies
Incompatible platelets transfused into patients with
high titre anti-A or anti-B have a decreased survival
Helen North
 patient
antibodies SPLEEN
Y
Y Y phagocytosis
YY Y
Y YY
Y

removal from
circulation

transfused
platelets complement and Immunoglobulin
Fc Receptors
Helen North
 Transfusion with non leucodepleted blood
History of pregnancy
Multiple transfusions

Seftel et al 2004; Blood 103(1) : 333-339

Helen North
 Role of H&I laboratory
Investigate for immune refractoriness
Identify the HLA and HPA antibody specificity
HLA (HPA) typing of patients
Maintain a panel of HLA and HPA typed donors
Select and issue HLA compatible platelets for
immune refractory patients
Essential to evaluate post-transfusion increments
Advice on patient management

Helen North
 Laboratory tests prior to provision of
HLA selected platelets
HLA class I typing
1x 6ml EDTA anti-coagulated sample
HLA-A, B,C type (HLA class I type)
using PCR-SSP or luminex SSO technology
Antigen level of typing
HLA antibody identification
1x 6ml clotted serum sample
Luminex technology to identify antibody specificity

Helen North
 Provision of HLA Selected
Platelets (HSP)
The provision of HSP is based on the
patient s:-
HLA type
HLA class I antibody specificity.
If present, HPA and ABO antibody specificity
If requested, CMV and RhD status is
taken into consideration.

Helen North
 Jan 2012

Alleles Antigens

HLA-A 1,729 28
HLA-B 2,329 61
HLA-C 1,291 10

To provide the best matched unit:

- Require at least 24 hours notice for routine orders of
HLA selected platelets

Helen North
 HLA selected platelets
Apheresis collection from a single donor
Platelet count >240x109/dose
Leucocyte depleted
Leucocyte count <5x106/dose
Irradiated
Known HLA and HPA genotype
In a stable patient the expected
increment after 1 dose = 30-40 x 109/L
Helen North
 The donor and patient are matched at the
antigen level of the HLA-A and B loci.

Patient A*01,A*02; B*08,B*44

Donor 1 A*01,A*02; B*08,B*44

Donor 2* A*01,A*01; B*08,B*08

*homozygous donor
Helen North
 B match grades (B1 B2 B3 B4)
Anywhere from one to four mismatched HLA
antigens (HLA-A and/or B loci) are transfused
into the patient.
Avoid mismatched antigen(s) to which the patient has
antibodies against
ideally the mismatched antigen(s) will be similar to the
patient s HLA antigens (reduces risk of further
sensitisation).
Patient s type: HLA-A1,A2;B8,B44
Donor s type: HLA-A1,A68*;B8,B44 (B1 match)
* = cross-reactive with HLA-A2

Helen North
Helen North
 Availability of HLA selected
platelets
14,000 typed platelet apheresis donors
Currently ~80% of platelet stock are from
apheresis donors
Patients with common HLA types
a good chance of finding a well matched platelet
from the daily platelet stock

Helen North
 2013 - HLA selected platelet units
In 2013:
15,783 HLA selected platelets issued by NHSBT
8,773 issued to London and SE region (55.6%)
Tooting and Colindale H&I lab

75% A and B1 match grade units (Colindale

2013)

Helen North
 Value of increments to NHSBT
To ensure that NHSBT is providing selected
platelets that are beneficial
Patients are reviewed weekly
If post transfusion increments are poor
(<10x109):
Investigate further cause of poor increments
presence of anti-HPA or ABO antibodies.
Patients actively being transfused:
Monitor their antibody profile monthly
Ensure no further sensitisation

Helen North
 For difficult to match patients:
Not all HLA antibodies cause refractoriness
Identify acceptable mismatches
Currently only receive 50% of increments
from hospitals
Varies greatly between hospitals

Helen North
 Current clinical trial:
Epitope matched HLA selected platelets

Currently HSP are matched based on antigens

However, antibodies bind to epitopes
HLAMatchmaker (Duquesnoy) identifies eplets
within antigens thought to represent epitopes

Helen North
 Current match Eplet match and priority
grade and priority
1st choice G052514302071N G072414175858*
A Grade (19 Eplet) (1 Eplet)
2nd choice G072414175973U G072414175973U
A Grade (17 Eplet) (17 Eplet)
3rd choice G072414175858* G052514302071N
B2 Grade (1 Eplet) (19 Eplet)

Data from Collette Pigden

Helen North
 Increase the number of acceptable
mismatched units
Without further sensitisation
Possibly increase our knowledge of
acceptable mismatching
Still recruiting patients to enrol in the trial

Helen North
 Case 1: Platelet refractoriness due to
HLA antibodies
Patient:: 54 yr old male, diagnosis: Myelodysplasia
Referred to H&I Colindale:
Poor response to ABO compatible pooled and
single donor platelets
HLA type HLA-A*30, A*32; B*18, B*35
HLA antibodies detected:
All HLA-A locus antigens except A30 and A32
B7 CREG antigens
(B7,27,40,42,48,54,55,56,73,81)
Also B8,B44,B45,B57,B58

Helen North
 Case 1
Due to the patient's antibody profile:
predicted not to increment with over 90% of
random platelets
Pre transfusion count: 13 x 109/L
Post transfusion count on random platelets:
18 x 109/L

Helen North
 Case 1: Increments following HLA
selected platelets

Patient's HLA type is A30,A32;B18,B35

A match grade HLA selected platelets
incremented from 16 to 43 x109/L
However, only 5 A matched donors
available in the UK
B2 match grade HLA selected platelets
A32,X;B14,B52 incremented from 24 to 43
x109/L
Helen North
 Patient management
Patient with MDS requires lifelong platelet support
As an outpatient, requires two HLA selected
platelet units/week
to maintain an adequate platelet count.
As only a small selection of donors are suitable
Search for suitably matched donors (A, B1,B2
match) and arrange for them to be bled to meet
expected requests.
Difficult to manage this patient at short notice

Helen North