Personalized peptide arrays for detection of HLA alloantibodies in organ transplantation

Pan Liu, Tomokazu Souma, Andrew Zu Sern Wei, Xueying Xie, Xunrong Luo, Jing Jin*

*Corresponding author for this work

Research output: Contribution to journalArticle

Abstract

In organ transplantation, the function and longevity of the graft critically rely on the success of controlling immunological rejection reactivity against human leukocyte antigens (HLA). Histocompatibility guidelines are based on laboratory tests of anti-HLA immunity, which presents either as pre-existing or de novo generated HLA antibodies that constitute a major transplantation barrier. Current tests are built on a single-antigen beads (SAB) platform using a fixed set of ~100 preselected recombinant HLA antigens to probe transplant sera. However, in humans there exist a far greater variety of HLA types, with no two individuals other than identical twins who can share the same combination of HLA sequences. While advanced technologies for HLA typing and direct sequencing can precisely capture any mismatches in DNA sequence between a donor’s and recipient’s HLA, the SAB assay, due to its limited variety in sequence representation, is unable to precisely detect alloantibodies specifically against the donor HLA mismatches. We sought to develop a complementary method using a different technology to detect and characterize antidonor HLA antibodies on a personalized basis. The screening tool is a custom peptide array of donor HLA-derived sequences for probing posttransplant sera of the organ recipient to assess the risk for antibody-mediated rejection. On a single array for one donor-recipient pair, up to 600 unique peptides are made based on the donor’s HLA protein sequences, each peptide carrying at least one mismatched residue in a 15-amino acid sequence. In our pilot experiments to compare antigen patterns for pre- and post-transplant sera on these arrays, we were able to detect anti-HLA signals with the resolution that also allowed us to pinpoint the immune epitopes involved. These personalized antigen arrays allow highresolution detection of donor-specific HLA epitopes in organ transplantation.

Original languageEnglish (US)
Article numbere56278
JournalJournal of Visualized Experiments
Volume2017
Issue number127
DOIs
StatePublished - Sep 6 2017

Fingerprint

Transplantation (surgical)
Isoantibodies
Organ Transplantation
Antigens
HLA Antigens
Peptides
Transplants
Tissue Donors
Antibodies
Epitopes
Serum
Technology
Histocompatibility
Monozygotic Twins
DNA sequences
Grafts

Keywords

  • Alloantibody
  • Antibody mediated transplant rejection (AMR)
  • Antigen array
  • Biochemistry
  • HLA mismatch
  • Human leukocyte antigen (HLA)
  • Issue 127
  • Organ transplantation
  • SPOT synthesis

ASJC Scopus subject areas

  • Neuroscience(all)
  • Chemical Engineering(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

Cite this

Liu, Pan ; Souma, Tomokazu ; Wei, Andrew Zu Sern ; Xie, Xueying ; Luo, Xunrong ; Jin, Jing. / Personalized peptide arrays for detection of HLA alloantibodies in organ transplantation. In: Journal of Visualized Experiments. 2017 ; Vol. 2017, No. 127.
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abstract = "In organ transplantation, the function and longevity of the graft critically rely on the success of controlling immunological rejection reactivity against human leukocyte antigens (HLA). Histocompatibility guidelines are based on laboratory tests of anti-HLA immunity, which presents either as pre-existing or de novo generated HLA antibodies that constitute a major transplantation barrier. Current tests are built on a single-antigen beads (SAB) platform using a fixed set of ~100 preselected recombinant HLA antigens to probe transplant sera. However, in humans there exist a far greater variety of HLA types, with no two individuals other than identical twins who can share the same combination of HLA sequences. While advanced technologies for HLA typing and direct sequencing can precisely capture any mismatches in DNA sequence between a donor’s and recipient’s HLA, the SAB assay, due to its limited variety in sequence representation, is unable to precisely detect alloantibodies specifically against the donor HLA mismatches. We sought to develop a complementary method using a different technology to detect and characterize antidonor HLA antibodies on a personalized basis. The screening tool is a custom peptide array of donor HLA-derived sequences for probing posttransplant sera of the organ recipient to assess the risk for antibody-mediated rejection. On a single array for one donor-recipient pair, up to 600 unique peptides are made based on the donor’s HLA protein sequences, each peptide carrying at least one mismatched residue in a 15-amino acid sequence. In our pilot experiments to compare antigen patterns for pre- and post-transplant sera on these arrays, we were able to detect anti-HLA signals with the resolution that also allowed us to pinpoint the immune epitopes involved. These personalized antigen arrays allow highresolution detection of donor-specific HLA epitopes in organ transplantation.",
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Personalized peptide arrays for detection of HLA alloantibodies in organ transplantation. / Liu, Pan; Souma, Tomokazu; Wei, Andrew Zu Sern; Xie, Xueying; Luo, Xunrong; Jin, Jing.

In: Journal of Visualized Experiments, Vol. 2017, No. 127, e56278, 06.09.2017.

Research output: Contribution to journalArticle

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T1 - Personalized peptide arrays for detection of HLA alloantibodies in organ transplantation

AU - Liu, Pan

AU - Souma, Tomokazu

AU - Wei, Andrew Zu Sern

AU - Xie, Xueying

AU - Luo, Xunrong

AU - Jin, Jing

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AB - In organ transplantation, the function and longevity of the graft critically rely on the success of controlling immunological rejection reactivity against human leukocyte antigens (HLA). Histocompatibility guidelines are based on laboratory tests of anti-HLA immunity, which presents either as pre-existing or de novo generated HLA antibodies that constitute a major transplantation barrier. Current tests are built on a single-antigen beads (SAB) platform using a fixed set of ~100 preselected recombinant HLA antigens to probe transplant sera. However, in humans there exist a far greater variety of HLA types, with no two individuals other than identical twins who can share the same combination of HLA sequences. While advanced technologies for HLA typing and direct sequencing can precisely capture any mismatches in DNA sequence between a donor’s and recipient’s HLA, the SAB assay, due to its limited variety in sequence representation, is unable to precisely detect alloantibodies specifically against the donor HLA mismatches. We sought to develop a complementary method using a different technology to detect and characterize antidonor HLA antibodies on a personalized basis. The screening tool is a custom peptide array of donor HLA-derived sequences for probing posttransplant sera of the organ recipient to assess the risk for antibody-mediated rejection. On a single array for one donor-recipient pair, up to 600 unique peptides are made based on the donor’s HLA protein sequences, each peptide carrying at least one mismatched residue in a 15-amino acid sequence. In our pilot experiments to compare antigen patterns for pre- and post-transplant sera on these arrays, we were able to detect anti-HLA signals with the resolution that also allowed us to pinpoint the immune epitopes involved. These personalized antigen arrays allow highresolution detection of donor-specific HLA epitopes in organ transplantation.

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KW - Issue 127

KW - Organ transplantation

KW - SPOT synthesis

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