HLA 교차시험에서 AHG-CDC의 유세포 측정: 예비연구 (Flow Cytometric AHG-CDC for HLA Crossmatch: A Pilot Study)

서지정보

· 발행기관 : 대한수혈학회
· 수록지 정보 : 대한수혈학회지 / 21권 / 2호 / 122 ~ 131페이지
· 저자명 : 원동일

초록

배경: 장기이식을 위한 HLA 교차시험에서 보체의존세포독성법(CDC)은 기증자특이 HLA 항체의 존재와 보체고정 능력을 평가하는데 매우유용한 방법이다. 본 예비연구에서는 anti-human globulin (AHG)-증강 CDC (AHG-CDC) 반응 유발후 죽은 세포를 유세포분석법(flow cytometry, FC)으로 측정할 수 있는지 조사하였다(‘FC AHGCDC’).
방법: 본 FC AHG-CDC는 삼색상 유세포분석법으로서, 7-aminoactinomycin D로 염색하여 죽은세포 비율(% dead cells)을 측정하였다. flow cytometry crossmatch (FCXM) 총 45예(양성 12예, 음성 33예)에서 FC AHG-CDC를 시행하여 비교하였다.
결과: FC AHG-CDC의 % dead cells는 FCXM의mean fluorescent intensity ratio와 유의한 상관관계가 있었다(T세포: r=0.613, P=7.45×10−6; B세포:r=0.404, P=0.006). FCXM 양성예중 FC AHG-CDC 양성률은 T, B세포 각각 80% (8/10), 75% (9/12)로비교적 높았다. FCXM 음성예중 FC AHG-CDC 음성률은 T, B세포 각각 100% (35/35), 91%(30/33)이었다.
결론: 본 예비연구에서, FC AHG-CDC는 T와 B 세포의 물리적 분리와 혈청 배수 희석 없이도 각세포에 대한 보체고정 세포독성 항체의 농도에비례하는 정량적인 측정치인 % dead cells을 얻을수 있었다.

영어초록

Background: For HLA crossmatch in organ transplantation, complement-dependent cytotoxicity (CDC) is a very useful methodology to detect donor-specific HLA antibodies and their complement fixing ability. In this preliminary pilot study, we investigated whether dead cells induced in anti-human globulin (AHG)-augmented CDC (AHG-CDC) reaction could be measured by flow cytometry (‘FC AHG-CDC’).
Methods: This FC AHG-CDC measured the percentage of dead cells (% dead cells) after 7-aminoactinomycin D staining using 3 color flow cytometry. A total 45 flow cytometry crossmatch (FCXM) cases of 12 positives and 33 negatives were further tested using this FC AHG-CDC.
Results: The % dead cells of FC AHG-CDC was significantly correlated with the mean fluorescent intensity ratio of FCXM (T cells: r=0.613, P=7.45×10-6; B cells: r=0.404, P=0.006). The positivity rate of FC AHG-CDC among FCXM positive cases was relatively high: 80% (8/10) for T cells and 75% (9/12) for B cells. The negativity rate of FC AHG-CDC among FCXM negative cases was 100% (35/35) for T cells and 91% (30/33)for B cells.
Conclusion: In this pilot study, FC AHG-CDC could yield quantitative values, % dead cells, which was proportional to the level of complement-fixing cytotoxic antibodies against T and B cells, respectively, even without physical separation of cells or serial dilution of serum. (Korean J Blood Transfus 2010;21:122-131)

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