器官移植之流式細胞儀交叉配對試驗方法的建立

交叉配對試驗是器官移植前必須進行的檢驗，而流式細胞儀交叉配對法（flow cytometric cross-matching，FCXM）是目前最敏感的方法。本研究為建立FCXM方法，共收集11對欲進行器官移植之捐贈者和受贈者血液檢體，以流式細胞分析技術檢測受贈者血清抗體與捐贈者T細胞和B細胞的反應，並將其結果與傳統的補體依賴性細胞毒殺法（complement-dependent cytotoxicity，CDC）進行比較。其中，第1－7例檢體的結果顯示：T細胞之配對試驗可清楚呈現陽性與陰性結果，而B細胞因為發生非特異性反應而無法明確判讀。因此，針對第8－11例檢體，增加捐贈者細胞以pronase酵素進行前處理，此步驟對T細胞反應無影響，而可以解決B細胞高背景值問題，並提高FCXM方法的特異性與敏感度。此11例檢體中，9例FCXM法的結果與CDC法一致，然而，有2例檢體在兩種方法的結果不相符合，因此也進一步探討其可能的原因。

關鍵字

交叉配對試驗；流式細胞分析技術； T細胞； B細胞； pronase酵素

並列摘要

Crossmatching is an essential procedure to determine preformed anti-donor HLA antibodies in recipient sera before organ transplantation, and the assay using flow cytometry is currently recognized as the most sensitive method. To establish flow cytometric crossmatching (FCXM) at our laboratory, blood samples collected from 11 pairs of donors and recipients were used to setup the protocol. Meanwhile, the results from FCXM were compared with that from complement- dependent cytotoxicity (CDC) method. In the first 7 cases, as to T cell FCXM (T-FXCM), positive and negative results could be clearly distinguished. However, in B cell FCXM (B-FCXM), the interpretation was inconclusive and uncomfortable because of B cells’ non-specific reaction with serum antibodies. Therefore, in the cases 8-11, we pretreated donor cells with pronase enzyme to remove Fc receptors on cell surfaces before FCXM assay. The pronase treatment had no apparent effect on T-FCXM, but it could dramatically resolve the high background problem on B-FCXM and further improve both the specificity and sensitivity of FCXM. Nine out of 11 cases demonstrated the same results in FCXM and CDC assays. However, incompatible results were observed in 2 cases and the possible causes were further discussed.

並列關鍵字

Crossmatching ； flow cytometry ； T cell ； B cell ； pronase

國際替代計量

器官移植之流式細胞儀交叉配對試驗方法的建立

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