Buabut B, Chiewsilp P, Patanapanyasat K, Jirasiritham S, Mavichak V, Sujirachato K, Mongkolsuk T
Department of Pathology, Pramongkutklao Hospital, Bangkok, Thailand.
J Med Assoc Thai. 1997 Sep;80 Suppl 1:S55-61.
Accelerated acute cellular rejection (AR) continues to be a serious problem in kidney transplantation (KT), suggesting that undetected presensitization may be encountered. The purpose of this study was to determine the most sensitive crossmatching (XM) technique to detect the preformed antibody (Ab) which may cause AR. One hundred and twenty two sera from 98 patients, on the waiting list for KT at Ramathibodi Hospital were XMed with 23 cadaveric splenic lymphocytes including 2 living related KT (LR-KT). The XM was performed by 3 different techniques namely, standard microlymphocytotoxicity test (standard NIH), antihuman globulin microlymphocytotoxicity test (AHG) and flow cytometric XM (FCXM). The XM results revealed that 8 out of 75 (10.7%) tests were negative by standard NIH, i.e., 5 tests were positive by AHG only and 1 test was positive by FCXM only and 2 tests were positive by both AHG and FCXM. In addition, the patients who had the AHG technique were not done, 5 out of 47 (10.7%) tests were also negative by standard NIH but were positive by FCXM. The sensitivity of the techniques was done by titrations of anti HLA-A2. It was found that FCXM was the most sensitive technique, followed by AHG and standard NIH, consecutively. In the retrospective study of LR-KT, case #1, the standard NIH for XM using pre-KT blood sample was negative while AHG and FCXM were strongly positive. The patient had AR at day 2 post-KT which confirmed by needle biopsy. The serum at day 11 and day 116 post-KT were tested again and were positive by the 3 techniques. Case #2, pre-KT blood sample showed negative T-XM by the 3 techniques while auto-B and B-XM were positive by standard NIH and AHG but negative by FCXM. This patient had rejection at day 16 after KT. The post-KT blood sample at day 30 showed positive auto T/B and T/B-XM by standard NIH and AHG whereas it was still negative by FCXM. It was also noted that Ab to donor B cell was better detected by standard NIH and AHG than FCXM. In conclusion, FCXM is more sensitive than standard NIH and AHG, however this technique is limited in detecting IgM T and B cell Ab. AHG technique can detect both IgG and IgM antidonor T and B cell Abs. In addition, AHG technique is more sensitive than standard NIH and does not require sophisticated equipment. AHG technique should be appropriate for routine XM, especially, in LR-KT and sensitized patients.
加速性急性细胞排斥反应(AR)仍是肾移植(KT)中的一个严重问题,这表明可能存在未被检测到的预致敏情况。本研究的目的是确定最敏感的交叉配型(XM)技术,以检测可能导致AR的预先形成的抗体(Ab)。对拉玛蒂博迪医院等待肾移植的98例患者的122份血清,与23份尸体脾脏淋巴细胞进行交叉配型,其中包括2例活体亲属肾移植(LR-KT)。交叉配型采用3种不同技术,即标准微量淋巴细胞毒性试验(标准NIH法)、抗人球蛋白微量淋巴细胞毒性试验(AHG法)和流式细胞术交叉配型(FCXM法)。交叉配型结果显示,75次检测中有8次(10.7%)标准NIH法结果为阴性,即5次仅AHG法为阳性,1次仅FCXM法为阳性,2次AHG法和FCXM法均为阳性。此外,未进行AHG技术检测的患者中,47次检测中有5次(10.7%)标准NIH法结果也为阴性,但FCXM法为阳性。通过抗HLA-A2滴定来确定这些技术的敏感性。结果发现,FCXM法是最敏感的技术,其次是AHG法和标准NIH法。在LR-KT的回顾性研究中,病例1,肾移植前血样的标准NIH交叉配型为阴性,而AHG法和FCXM法均为强阳性。该患者在肾移植术后第2天发生AR,经穿刺活检证实。肾移植术后第11天和第116天的血清再次检测,3种技术均为阳性。病例2,肾移植前血样3种技术的T-XM均为阴性,而自身B细胞和B-XM标准NIH法和AHG法为阳性,但FCXM法为阴性。该患者在肾移植术后第16天发生排斥反应。肾移植术后第30天的血样标准NIH法和AHG法的自身T/B细胞和T/B-XM为阳性,而FCXM法仍为阴性。还注意到,标准NIH法和AHG法比FCXM法能更好地检测到针对供体B细胞的抗体。总之,FCXM法比标准NIH法和AHG法更敏感,然而该技术在检测IgM T细胞和B细胞抗体方面存在局限性。AHG技术可检测IgG和IgM抗供体T细胞和B细胞抗体。此外,AHG技术比标准NIH法更敏感,且不需要复杂设备。AHG技术适用于常规交叉配型,尤其是在LR-KT和致敏患者中。