Prediction of crossmatch outcome of highly sensitized patients by single and/or multiple antigen bead luminex assay

Smita Vaidya, David Partlow, Brian Susskind, Maryam Noor, Titus Barnes, Kristene Gugliuzza

Research output: Contribution to journalArticle

75 Citations (Scopus)

Abstract

BACKGROUND. We used a solid-phase assay to identify human leukocyte antigen (HLA) Class I and II specificities in highly reactive sera, and applied this information to predict crossmatch outcome with greater than 90% accuracy. METHODS. Sera from 20 highly sensitized end-stage renal disease patients reactive to 70-100% of HLA Class I and II antigen panel were analyzed by single and/or multiple antigen flow microparticle bead assay using Luminex platform (Luminex assay). These sera contained antibodies directed against high frequency public HLA class I and/or II epitopes accounting for 70-100% of serum's total reactivity. More than 2,000 complement dependent cytotoxicity (CDC) and 200 flow crossmatches (FLXM) were performed using sera from these patients and deceased donor T and B lymphocytes. RESULTS. Luminex serum analysis was able to correctly predict outcomes of 95% of T and B cell FLXM. In contrast, predictive values for the CDC T and B cell crossmatches by Luminex serum analysis were only 77% and 75%, respectively. The use of serum analysis in donor selection would have reduced the total number of required FLXM by more than 50%. The frequency of negative T cell FLXM was 56% when donors were selected randomly; however, if serum antibody analysis had been used to preselect the donors by excluding donors that were likely to be positive, the frequency of corresponding negative crossmatches would have increased up to 93%. CONCLUSION. This approach to donor selection may allow wider geographic sharing of cadaver donor organs without actually performing the crossmatch.

Original languageEnglish (US)
Pages (from-to)1524-1528
Number of pages5
JournalTransplantation
Volume82
Issue number11
DOIs
StatePublished - Dec 2006

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Antigens
Serum
Tissue Donors
HLA Antigens
T-Lymphocytes
Donor Selection
B-Lymphocytes
Complement C2
Histocompatibility Antigens Class I
Antibodies
Histocompatibility Antigens Class II
Cadaver
Chronic Kidney Failure
Epitopes

Keywords

  • Complement dependent cytotoxicity crossmatch
  • Flow cytometry crossmatch

ASJC Scopus subject areas

  • Transplantation
  • Immunology

Cite this

Prediction of crossmatch outcome of highly sensitized patients by single and/or multiple antigen bead luminex assay. / Vaidya, Smita; Partlow, David; Susskind, Brian; Noor, Maryam; Barnes, Titus; Gugliuzza, Kristene.

In: Transplantation, Vol. 82, No. 11, 12.2006, p. 1524-1528.

Research output: Contribution to journalArticle

Vaidya, Smita ; Partlow, David ; Susskind, Brian ; Noor, Maryam ; Barnes, Titus ; Gugliuzza, Kristene. / Prediction of crossmatch outcome of highly sensitized patients by single and/or multiple antigen bead luminex assay. In: Transplantation. 2006 ; Vol. 82, No. 11. pp. 1524-1528.
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abstract = "BACKGROUND. We used a solid-phase assay to identify human leukocyte antigen (HLA) Class I and II specificities in highly reactive sera, and applied this information to predict crossmatch outcome with greater than 90{\%} accuracy. METHODS. Sera from 20 highly sensitized end-stage renal disease patients reactive to 70-100{\%} of HLA Class I and II antigen panel were analyzed by single and/or multiple antigen flow microparticle bead assay using Luminex platform (Luminex assay). These sera contained antibodies directed against high frequency public HLA class I and/or II epitopes accounting for 70-100{\%} of serum's total reactivity. More than 2,000 complement dependent cytotoxicity (CDC) and 200 flow crossmatches (FLXM) were performed using sera from these patients and deceased donor T and B lymphocytes. RESULTS. Luminex serum analysis was able to correctly predict outcomes of 95{\%} of T and B cell FLXM. In contrast, predictive values for the CDC T and B cell crossmatches by Luminex serum analysis were only 77{\%} and 75{\%}, respectively. The use of serum analysis in donor selection would have reduced the total number of required FLXM by more than 50{\%}. The frequency of negative T cell FLXM was 56{\%} when donors were selected randomly; however, if serum antibody analysis had been used to preselect the donors by excluding donors that were likely to be positive, the frequency of corresponding negative crossmatches would have increased up to 93{\%}. CONCLUSION. This approach to donor selection may allow wider geographic sharing of cadaver donor organs without actually performing the crossmatch.",
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N2 - BACKGROUND. We used a solid-phase assay to identify human leukocyte antigen (HLA) Class I and II specificities in highly reactive sera, and applied this information to predict crossmatch outcome with greater than 90% accuracy. METHODS. Sera from 20 highly sensitized end-stage renal disease patients reactive to 70-100% of HLA Class I and II antigen panel were analyzed by single and/or multiple antigen flow microparticle bead assay using Luminex platform (Luminex assay). These sera contained antibodies directed against high frequency public HLA class I and/or II epitopes accounting for 70-100% of serum's total reactivity. More than 2,000 complement dependent cytotoxicity (CDC) and 200 flow crossmatches (FLXM) were performed using sera from these patients and deceased donor T and B lymphocytes. RESULTS. Luminex serum analysis was able to correctly predict outcomes of 95% of T and B cell FLXM. In contrast, predictive values for the CDC T and B cell crossmatches by Luminex serum analysis were only 77% and 75%, respectively. The use of serum analysis in donor selection would have reduced the total number of required FLXM by more than 50%. The frequency of negative T cell FLXM was 56% when donors were selected randomly; however, if serum antibody analysis had been used to preselect the donors by excluding donors that were likely to be positive, the frequency of corresponding negative crossmatches would have increased up to 93%. CONCLUSION. This approach to donor selection may allow wider geographic sharing of cadaver donor organs without actually performing the crossmatch.

AB - BACKGROUND. We used a solid-phase assay to identify human leukocyte antigen (HLA) Class I and II specificities in highly reactive sera, and applied this information to predict crossmatch outcome with greater than 90% accuracy. METHODS. Sera from 20 highly sensitized end-stage renal disease patients reactive to 70-100% of HLA Class I and II antigen panel were analyzed by single and/or multiple antigen flow microparticle bead assay using Luminex platform (Luminex assay). These sera contained antibodies directed against high frequency public HLA class I and/or II epitopes accounting for 70-100% of serum's total reactivity. More than 2,000 complement dependent cytotoxicity (CDC) and 200 flow crossmatches (FLXM) were performed using sera from these patients and deceased donor T and B lymphocytes. RESULTS. Luminex serum analysis was able to correctly predict outcomes of 95% of T and B cell FLXM. In contrast, predictive values for the CDC T and B cell crossmatches by Luminex serum analysis were only 77% and 75%, respectively. The use of serum analysis in donor selection would have reduced the total number of required FLXM by more than 50%. The frequency of negative T cell FLXM was 56% when donors were selected randomly; however, if serum antibody analysis had been used to preselect the donors by excluding donors that were likely to be positive, the frequency of corresponding negative crossmatches would have increased up to 93%. CONCLUSION. This approach to donor selection may allow wider geographic sharing of cadaver donor organs without actually performing the crossmatch.

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