HLA Class II 항체 검출 시 C3d 검사법과 SAB 검사법의 상관관계 평가
Correlation Between C3d Assay and Single Antigen Bead Assay for Detection of Human Leukocyte Antigen Class II Antibodies
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배경: 장기이식 전에 공여자 선택을 위해 사전에 공여자 HLA에 대한 항체 유무를 확인하는 것이 중요하고, 이식 후의 급성 또는 만성 거부반응을 예측하기 위해서도 HLA 항체에 대한 주기적인 추적 검사가 필요하다. 본 연구에서는 보체 활성화를 유발하는 HLA 항체를 검출할 수 있는 Luminex 검사법 중 C3d 검사를 시행하여 기존의 SAB 검사법과 비교하였다. 방법: HLA 항체 검사를 위해 검사실로 의뢰된 검체 중 class II SAB 검사(Immucor Transplant Diagnostics, USA)에서 양성인 혈액 검체 중 최종적으로 DSA 양성인 66개의 검체를 대상으로 하였다. 각 검체를 SAB 검사법과 C3d 검사법(Immucor Transplant Diagnostics)으로 검사를 시행한 후 MATCH IT! Antibody v1.3.1.5 (Immucor Transplant Diagnostics) 프로그램으로 HLA 항체 종류, MFI 값, 양성여부를 파악한 후 두 검사의 상관관계를 분석하였다. 결과: SAB 검사의 항체수는 11-20개인 구간에서 24개(36.4%)로 가장 많았고, C3d 검사는 항체수가 2-5개인 구간에서 23개(34.8%)로 가장 많았다. SAB 검사에서 양성인 항체 중에서 MFI≤3,000인 항체 440개 중 28개(6.4%)만이 C3d 검사 양성이었고, MFI≥3,001인 항체 556개 중 341개(61.3%)가 C3d 검사 양성이었다. 반면, C3d 검사에서 양성인 442개의 항체 중에서는 MFI≤500, 1,001≤MFI≤10,000인 구간에서 각각 32개(7.2%), 41개(9.3%)를 제외하고는 모두 SAB 검사도 양성이었다. C3d 검사 양성 검체가 C3d 검사 음성 검체보다 SAB의 최대 MFI 값이 더 큰 것으로 나타났다. 결론: HLA class II 항체의 SAB 검사에서의 MFI 값은 C3d 검사 음성인 경우보다 양성인 경우에 더 큰 것으로 나타났으며, C3d 검사법의 임상적 중요성에 대해서는 추가적인 연구가 필요할 것이다.
Abstract
Background: Detection of anti-human leukocyte antigen (HLA) antibodies is important during the selection of an appropriate donor prior to organ transplantation and also for monitoring the patients after transplantation. In this study, we compared antibodies detected via C3d assays, which monitors C3d complement-binding activities of HLA antibodies with those detected via single antigen bead (SAB) assays. Methods: A total of 66 serum samples were tested in parallel by SAB assays (Immucor Transplant Diagnostics, USA) and C3d assays (Immucor) for the detection of HLA class II antibodies. The relationship between these two methods was analyzed based on the types, numbers, median fluorescent intensity (MFI) values, and positivity of the antibodies using MATCH IT! Antibody (Immucor) program. Results: The number of antibodies obtained based on SAB and C3d assays was the highest with 24 samples (36.4%) in the 11–20 range and 23 (34.8%) in the 2–5 range detected via each assay. Among the SAB-positive antibodies, only 28 (6.4%) of the 440 antibodies with MFI ≤3,000 were C3d-positive, and 341 (61.3%) of the 556 antibodies with MFI ≥3,001 were C3d-positive. Whereas, among the 442 C3d-positive antibodies, SAB assays were positive except for 32 (7.2%) and 41 (9.3%) antibodies in the sections of MFI ≤500 and 1,001 ≤MFI ≤10,000, respectively. C3d-positive samples had higher maximum MFI values based on SAB assays, compared with C3d-negative samples. Conclusions: MFI values of HLA class II antibodies detected through SAB assays in C3d-positive samples were higher than those in C3d-negative samples.
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