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Graft Immunocomplex Capture Fluorescence Analysis to Detect Donor-Specific Antibodies and HLA Antigen Complexes in the Allograft.
Nakamura, Tsukasa; Ushigome, Hidetaka; Watabe, Kiyoko; Imanishi, Yui; Masuda, Koji; Matsuyama, Takehisa; Harada, Shumpei; Koshino, Katsuhiro; Iida, Taku; Nobori, Shuji; Yoshimura, Norio.
Afiliación
  • Nakamura T; a Department of Organ Transplantation and General Surgery , Kyoto Prefectural University of Medicine , Kyoto , Japan.
  • Ushigome H; a Department of Organ Transplantation and General Surgery , Kyoto Prefectural University of Medicine , Kyoto , Japan.
  • Watabe K; b Department of Blood Transfusion and Cell Therapy , Kyoto Prefectural University of Medicine , Kyoto , Japan.
  • Imanishi Y; b Department of Blood Transfusion and Cell Therapy , Kyoto Prefectural University of Medicine , Kyoto , Japan.
  • Masuda K; a Department of Organ Transplantation and General Surgery , Kyoto Prefectural University of Medicine , Kyoto , Japan.
  • Matsuyama T; a Department of Organ Transplantation and General Surgery , Kyoto Prefectural University of Medicine , Kyoto , Japan.
  • Harada S; a Department of Organ Transplantation and General Surgery , Kyoto Prefectural University of Medicine , Kyoto , Japan.
  • Koshino K; a Department of Organ Transplantation and General Surgery , Kyoto Prefectural University of Medicine , Kyoto , Japan.
  • Iida T; a Department of Organ Transplantation and General Surgery , Kyoto Prefectural University of Medicine , Kyoto , Japan.
  • Nobori S; a Department of Organ Transplantation and General Surgery , Kyoto Prefectural University of Medicine , Kyoto , Japan.
  • Yoshimura N; a Department of Organ Transplantation and General Surgery , Kyoto Prefectural University of Medicine , Kyoto , Japan.
Immunol Invest ; 46(3): 295-304, 2017 Apr.
Article en En | MEDLINE | ID: mdl-28151033
BACKGROUND: Immunocomplex capture fluorescence analysis (ICFA) is an attractive method to detect donor-specific anti-HLA antibodies (DSA) and HLA antigen complexes. Currently, antibody-mediated rejection (AMR) due to DSA is usually diagnosed by C4d deposition and serological DSA detection. Conversely, there is a discrepancy between these findings frequently. Thereupon, our graft ICFA technique may contribute to establish the diagnosis of AMR. METHODS: Graft samples were obtained by a percutaneous needle biopsy. Then, the specimen was dissolved in PBS by the lysis buffer. Subsequently, HLA antigens were captured by anti-HLA beads. Then, DSA-HLA complexes were detected by PE-conjugated anti-human IgG antibodies, where DSA had already reacted with the allograft in vivo, analyzed by a Luminex system. RESULTS: A ratio (sample MFI/blank beads MFI) was calculated: ≥ 1.0 was determined as positive. We found that DSA-HLA complexes in the graft were successfully detected from only slight positive 1.03 to 79.27 in a chronic active AMR patient by graft ICFA. Next, positive graft ICFA had predicted the early phase of AMR (MFI ratio: 1.38) even in patients with no serum DSA. Finally, appropriate therapies for AMR deleted DSA deposition (MFI ratio from 0.3 to 0.7) from allografts. CONCLUSIONS: This novel application would detect early phase or incomplete pathological cases of AMR, which could lead to a correct diagnosis and initiation of appropriate therapies. Moreover, graft ICFA might address a variety of long-standing questions in terms of DSA. ABBREVIATIONS: AMR: Antibody-mediated rejection; DSA: Donor-specific antibodies; ICFA: Immunocomplex capture fluorescence analysis.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Técnica del Anticuerpo Fluorescente / Trasplante de Riñón / Rechazo de Injerto Tipo de estudio: Prognostic_studies Límite: Adult / Aged / Female / Humans / Male / Middle aged Idioma: En Revista: Immunol Invest Asunto de la revista: ALERGIA E IMUNOLOGIA Año: 2017 Tipo del documento: Article País de afiliación: Japón Pais de publicación: Reino Unido
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Técnica del Anticuerpo Fluorescente / Trasplante de Riñón / Rechazo de Injerto Tipo de estudio: Prognostic_studies Límite: Adult / Aged / Female / Humans / Male / Middle aged Idioma: En Revista: Immunol Invest Asunto de la revista: ALERGIA E IMUNOLOGIA Año: 2017 Tipo del documento: Article País de afiliación: Japón Pais de publicación: Reino Unido