Gao Su-Qing, Xu Yun-Ping, Luo Chang-Ru, Li Da-Cheng, Pen Long, Liu Tong, Zou Qiong-Cai
Shenzhen Blood Center Institute of Transfusion Medicine;Shenzhen 518040, Guangdong Province, China.E-mail:
Shenzhen Blood Center Institute of Transfusion Medicine;Shenzhen 518040, Guangdong Province, China.
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2024 Feb;32(1):242-249. doi: 10.19746/j.cnki.issn.1009-2137.2024.01.039.
To establish a graded method to avoid mean fluorescence intensity (MFI) threshold of HLA Class I antibodies corresponding antigen, and the HLAMatchmaker program has been used to select the minimum mismatch value of donor-patient epitopes. Evaluate the application value of combining both methods in selecting HLA compatible platelets (PTL) for patients with immune platelet transfusion failure (IPTR) in improving platelet the corrected count increment (CCI).
A total 7 807 PLT cross-matching compatible were performed by the solid-phase red cell adherence (SPRCA) method for 51 IPTR patients. The Luminex single antigen flow cytometry was used to detect HLA Class I antibodies in patients, and detected the MFI value for different specificity antigens of HLA Class I antibodies, was graded into strong positive group (MFI>4 000, level 1), medium positive group (1 000< MFI≤4 000, 2), weak positive group (500< MFI≤1 000, 3), and one negative control group (MFI≤500). The results of 7 807 SPRCA their negative/positive reaction wells were enrolled and statistically analyzed in different grades and the four groups, the statistical differences between the four groups were compared. Multiple applications for the select HLA Class I compatible donor events were made for patients in two cases, and HLAMatchmaker program was used to calculate the number of HLA Class I epitopes mismatches between the donors and patients. The donor with the minimum number of epitopes mismatches was selected, while avoiding the corresponding antigens of HLA Class I antibodies in levels 1 and 2, the provision of HLA compatible platelets for IPTR. After the transfusions, the CCI value of the platelet transfusion efficacy evaluation index was calculated, and the clinical evaluation of the transfusion effect was obtained through statistical analysis.
There were statistically significant differences in the positive results of SPRCA immunoassay among the strong positive group, medium positive group, and weak positive group of 51 IPTR patients with different specific of HLA -I class antibodies and corresponding antigens(all <0.001). The positive results showed a range from high to low, with strong positive group>medium positive group>weak positive group. There were a statistical difference among between the strongly positive or moderately positive groups and the negative control group( <0.001). There was no statistical difference between the weakly positive group and the negative control group( >0.05). The strong positive group was set as the corresponding specific HLA Class I site corresponding antigen grade 1 avoidance threshold, the medium positive group as the grade 2 avoidance thresholds, and the weak positive group as the grade 3 avoidance threshold. In the case of donor platelet shortage, it is not necessary to avoid the weak positive group. Avoiding the strategy of donor antigens and HLAMatchmaker program scores ≤7 corresponding to HLA Class I antibodies of levels 1 and 2, with CCI values>4.5×10/L within 24 hours, can obtain effective clinical platelet transfusion conclusions.
When selecting HLA Class I compatible donors for IPTR patients, the grading avoids HLA Class I antibodies corresponding to donor antigens, and the donor selection strategy with the minimum scores of HLAMatchmaker program is comprehensively selected. The negative result confirmed by platelet cross-matching experiments has certain practical application value for improving platelet count in IPTR patients.
建立一种分级方法以避免HLAⅠ类抗体对应抗原的平均荧光强度(MFI)阈值,采用HLAMatchmaker程序选择供者 - 患者表位的最小错配值。评估两种方法联合应用于为免疫性血小板输注无效(IPTR)患者选择HLA相合血小板(PTL)以提高血小板校正计数增加值(CCI)的应用价值。
采用固相红细胞吸附(SPRCA)法对51例IPTR患者进行了共7807次血小板交叉配型相合检测。采用Luminex单抗原流式细胞术检测患者的HLAⅠ类抗体,检测HLAⅠ类抗体不同特异性抗原的MFI值,分为强阳性组(MFI>4000,1级)、中度阳性组(1000<MFI≤4000,2级)、弱阳性组(500<MFI≤1000,3级)和一个阴性对照组(MFI≤500)。将7807次SPRCA结果及其阴性/阳性反应孔按不同分级和四组进行登记并统计分析,比较四组间的统计学差异。对两例患者多次进行选择HLAⅠ类相合供者事件,应用HLAMatchmaker程序计算供者与患者之间HLAⅠ类表位错配数。选择表位错配数最少的供者,同时避免1级和2级HLAⅠ类抗体对应的抗原,为IPTR患者提供HLA相合血小板。输血后,计算血小板输注疗效评价指标CCI值,通过统计分析获得输血效果的临床评价。
51例不同特异性HLA -Ⅰ类抗体及对应抗原的IPTR患者,其强阳性组、中度阳性组和弱阳性组SPRCA免疫分析阳性结果有统计学差异(均<0.001)。阳性结果呈从高到低的趋势,强阳性组>中度阳性组>弱阳性组。强阳性或中度阳性组与阴性对照组之间有统计学差异(<0.001)。弱阳性组与阴性对照组之间无统计学差异(>0.05)。将强阳性组设为相应特异性HLAⅠ类位点对应抗原1级避免阈值,中度阳性组设为2级避免阈值,弱阳性组设为3级避免阈值。在供者血小板短缺的情况下,不必避免弱阳性组。避免供者抗原与HLAMatchmaker程序评分≤7对应1级和2级HLAⅠ类抗体的策略,24小时内CCI值>4.5×10/L,可获得有效的临床血小板输注结论。
为IPTR患者选择HLAⅠ类相合供者时,分级避免供者抗原对应的HLAⅠ类抗体,并综合选择HLAMatchmaker程序评分最小的供者选择策略。血小板交叉配型实验确认的阴性结果对提高IPTR患者血小板计数有一定的实际应用价值。