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Calcium-dependent HLA-DQ epitope revealed by EDTA mediated inhibition of antibody reactions in the Luminex single antigen bead assay.
de Marco, Renato; Noronha, Isaú H; Bottino, Luiza Zainotti Miguel Fahur; Dos Silva, Alan Andrew Santos; Liwski, Robert; Gerbase-DeLima, Maria.
Afiliação
  • de Marco R; Immunogenetics Institute-IGEN, Associação Fundo de Incentivo à Pesquisa (AFIP), São Paulo, Brazil.
  • Noronha IH; Immunogenetics Institute-IGEN, Associação Fundo de Incentivo à Pesquisa (AFIP), São Paulo, Brazil.
  • Bottino LZMF; Immunogenetics Institute-IGEN, Associação Fundo de Incentivo à Pesquisa (AFIP), São Paulo, Brazil.
  • Dos Silva AAS; Immunogenetics Institute-IGEN, Associação Fundo de Incentivo à Pesquisa (AFIP), São Paulo, Brazil.
  • Liwski R; Department of Pathology and Laboratory Medicine, Dalhousie University, Halifax, Canada.
  • Gerbase-DeLima M; Immunogenetics Institute-IGEN, Associação Fundo de Incentivo à Pesquisa (AFIP), São Paulo, Brazil.
HLA ; 104(2): e15628, 2024 Aug.
Article em En | MEDLINE | ID: mdl-39132721
ABSTRACT
Complement mediated interference with the detection of antibodies targeting HLA is a known limitation of the single antigen bead (SAB) Luminex assay. Ethylenediaminetetraacetic acid (EDTA) is currently the serum treatment of choice in most histocompatibility laboratories to block complement activation by chelating calcium. The purpose of this study was to investigate a serum with an antibody reactivity to HLA-DQ6, 7, 8 and 9 molecules, in the Luminex SAB assay, that was inhibited by treatment with EDTA. Serum was from a 55-year-old highly sensitised female renal transplant candidate that contained, among others, antibodies to an epitope containing the 74EL eplet, shared by HLA-DQ6, DQ7, DQ8 and DQ9 molecules. Serum samples were treated with EDTA, dithiothreitol (DTT), or heat prior to testing by SAB assay. EDTA-treated serum was also tested after the addition of calcium chloride (CaCl2). HLA-DQ-specific antibodies were isolated by adsorption/elution method using three informative donor cells and were tested in the absence or presence of EDTA. The antibody reactivity against HLA-DQ6, DQ7, DQ8 and DQ9 in the SAB assay was significantly inhibited by treating serum and eluates with EDTA and was restored by addition of CaCl2. The study represents the first description of a calcium-dependent epitope in HLA molecules. The relevance of this finding is that the treatment of sera with EDTA could lead to false-negative reactions in the SAB assay, which may compromise virtual crossmatching.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Teste de Histocompatibilidade / Antígenos HLA-DQ / Cálcio / Ácido Edético / Epitopos Limite: Female / Humans / Middle aged Idioma: En Revista: HLA Ano de publicação: 2024 Tipo de documento: Article País de afiliação: Brasil País de publicação: Reino Unido
Texto completo
Adicionar na Minha BVS
Imprimir
XML
PubMed Links
Buscar no Google

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Teste de Histocompatibilidade / Antígenos HLA-DQ / Cálcio / Ácido Edético / Epitopos Limite: Female / Humans / Middle aged Idioma: En Revista: HLA Ano de publicação: 2024 Tipo de documento: Article País de afiliação: Brasil País de publicação: Reino Unido