【摘要】 目的:建立分级规避HLA抗体MFI阈值对应抗原方法，联合应用HLAMatchmaker表位计算法，选择供患者表位最小错配评分值，评估两种方法为免疫性血小板输注无效(Immune platelet transfusion refractoriness, IPTR)患者选择HLA相容性血小板供者，在提升血小板输注后校正增加值(CCI)的应用价值。方法:采用SPRCA法完成51例IPTR患者的7 807次血小板交叉配型实验，判断其免疫反应阴/阳性结果。采用Luminex单抗原流式微珠法检测患者的HLA-I类抗体，获得不同特异性抗体对应HLA-I类抗原MFI值，并将其分组及分级，强阳性组(MFI>4 000,1级)、中阳性组(1 000<MFI≤4 000,2级)、弱阳性组(500<MFI≤1 000,3级)和1个阴性对照组(MFI≤500),分组统计SPRCA实验在四组中免疫反应的阳性及阴性结果检出数，比较四组之间统计学差异。对2例患者多次申请血小板配型选择HLA相容性供者事件，采用HLAMatchmaker计算供患者HLA表位错配评分数，选择最小HLA表位错配评分数供者，同时规避1和2级HLA-I类抗体对应抗原的供者，为患者输注适合供者血小板，输注之后计算CCI值，统计临床输注效果。结果:51例IPTR患者的不同特异性HLA-I类抗体及对应抗原在强阳性、中阳性和弱阳性三组之间的SPRCA实验免疫反应阳性结果检出数均存在统计学差异(均P<0.001),阳性结果检出数呈现由高到低格局(强阳性组>中阳性组>弱阳性组)。强阳性和中阳性组与阴性对照组之间的SPRCA实验免疫反应阳性结果检出数存在统计学差异(P<0.001),弱阳性位组和阴性对照组之间的SPRCA实验免疫反应阳性结果检出数无统计学差异(P>0.05)。设置强阳性组为相应特异性HLA位点对应抗原1级规避阈值，中阳性组为2级规避阈值，弱阳性组为3级规避阈值，在供者血小板紧缺情况下，可以不需要规避弱阳性组。规避1和2级HLA-I类抗体对应供者抗原及选择HLAMatchmaker表位错配评分数≤7血小板供者策略，24 h内CCI值均>4.5×10~9/L,均可获得临床血小板输注有效。结论:在为IPTR患者选择HLA-I类相容性供者时，分级规避HLA-I类抗体对应供者抗原，综合选择供受者HLAMatchmaker表位错配评分数≤7,经血小板交叉配型实验确认为阴性结果的供者选择策略，对提升IPTR患者血小板计数具有一定实际应用价值。更多还原

【Abstract】 Objective: 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). Methods: A total 7 807 PLT cross-matching compatible were performed by the solidphase red cell adherence( SPRCA) method for 51 IPTR patients. The Luminex single antigen flow cytometry w as used to detect HLA Class I antibodies in patients,and detected the M FI value for different specificity antigens of HLA Class I antibodies,w as graded into strong positive group( M FI > 4 000,level 1),medium positive group( 1 000 < M FI ≤4 000,2),w eak positive group( 500 < M FI≤1 000,3),and one negative control group( M FI≤500). The results of7 807 SPRCA their negative/positive reaction w ells w ere enrolled and statistically analyzed in different grades and the four groups,the statistical differences betw een the four groups w ere compared. M ultiple applications for the select HLA Class I compatible donor events w ere made for patients in tw o cases,and HLAM atchmaker program w as used to calculate the number of HLA Class I epitopes mismatches betw een the donors and patients. The donor w ith the minimum number of epitopes mismatches w as selected,w hile 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 w as calculated,and the clinical evaluation of the transfusion effect w as obtained through statistical analysis. Results: There w ere statistically significant differences in the positive results of SPRCA immunoassay among the strong positive group,medium positive group,and w eak positive group of 51 IPTR patients w ith different specific of HLA-I class antibodies and corresponding antigens( all P < 0. 001). The positive results show ed a range from high to low,w ith strong positive group > medium positive group > w eak positive group. There w ere a statistical difference among betw een the strongly positive or moderately positive groups and the negative control group( P< 0. 001). There w as no statistical difference betw een the w eakly positive group and the negative control group( P >0. 05). The strong positive group w as set as the corresponding specific HLA Class I site corresponding antigen grade 1avoidance threshold,the medium positive group as the grade 2 avoidance thresholds,and the w eak positive group as the grade 3 avoidance threshold. In the case of donor platelet shortage,it is not necessary to avoid the w eak positive group.Avoiding the strategy of donor antigens and HLAM atchmaker program scores ≤ 7 corresponding to HLA Class I antibodies of levels 1 and 2,w ith CCI values > 4. 5 × 109/L w ithin 24 hours,can obtain effective clinical platelet transfusion conclusions. Conclusion: 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 w ith the minimum scores of HLAM atchmaker program is comprehensively selected. The negative result confirmed by platelet crossmatching experiments has certain practical application value for improving platelet count in IPTR patients.更多还原