Objective: To explore the the effects of 2-Me, DTT, papain, pineapple protease and ZZAP on the antigenicity of JMH antigen of human red blood cells (RBC) surface.
Methods: Firstly, human RBC were treated with 2-Me, DTT, pineapple protease, papain and ZZAP reagents, respectively. The antigenicity of JMH antigen on human RBC surface was detected and analyzed by flow cytometry.
Results: Flow cytometric analysis found that compared with level before treatment, the antigenicity of JMH antigen on RBC surface was significantly reduced after 2-Me treatment, the positive rate of JMH antigen: 69.5%±4.5% vs 56.5%±3.4% (t=12.44, P＜0.01); fluorescence intensity: 4906±317 vs 3003±165 (t=11.84, P＜0.01). The antigenicity of JMH antigen on RBC surface significantly increased after DTT treatment, showing the positive rate of JMH antigen: 61.7%±3.8% vs 75.5±4.9% (t=16.57, P＜0.01), fluorescence intensity: 4044±294 vs 4854±319 (t=15.46, P＜0.01). However, both bromelain and papain could significantly reduce the antigenicity of JMH antigen on the RBC surface, Bromelain: the positive rate of JMH antigen: 62.2%±3.8% vs 8.8%±1.2% (t=26.44, P＜0.01), fluorescence intensity: 4263±273 vs 1444±212 (t=19.27, P＜0.01); Papain: the positive rate of JMH antigen: 62.8%±3.6% vs 8.8%±1.5% (t=21.38, P＜0.01), fluorescence intensity: 4389±284 vs 1458±230 (t=17.49, P＜0.01). The flow cytometric analysis revealed that ZZAP treatment significantly reduced the antigenicity of JMH antigen on the RBC surface, the positive rate of JMH antigen: 62.2%±4.4% vs 48.2%±4.1% (t=14.87, P＜0.01), fluorescence intensity: 4106±263 vs 2063±175 (t=17.49, P＜0.01).
Conclusion: The treatment with 2-Me can reduce the antigenicity of JMH antigen on human RBC surface. The antigenicity of JMH antigen on human RBC surface increased after DTT treatment. The antigenicity of JMH antigen on human RBC surface significantly reduces after the treatment with pineapple protease or papain. ZZAP treatment can reduce the antigenicity of JMH antigen on the RBC surface.
题目: 2Me、DTT、ZAAP及酶等介质的处理对 红细胞JMH抗原的影响.
目的: 探讨血型鉴定、抗体鉴定及交叉配血过程中常用工具试剂2Me、DTT、木瓜蛋白酶、菠萝蛋白酶及ZZAP对人红细胞表面JMH抗原抗原性的影响.
方法: 首先分别用2-Me、DTT、菠萝蛋白酶、木瓜蛋白酶及ZZAP试剂对人红细胞进行处理，然后通过流式细胞术检测并分析处理前后人红细胞表面JMH抗原的变化.
结果: 流式细胞术分析发现，与未处理组相比，2-Me处理组人红细胞表面JMH抗原的抗原性显著减弱，抗原的阳性率：69.5%±4.5% vs 56.5%±3.4%（t=12.44，P＜0.01）；荧光强度：4906±317 vs 3003±165（t=11.84，P＜0.01）。与2-Me不同，DTT处理可显著提高红细胞表面JMH抗原的抗原性，抗原阳性率：61.7%±3.8% vs 75.5±4.9%（t=16.57，P＜0.01）；荧光强度：4044±294 vs 4854±319（t=15.46，P＜0.01）;而菠萝蛋白酶与木瓜蛋白酶均可显著减弱红细胞表面JMH抗原的抗原性。菠萝蛋白酶处理后抗原阳性率：62.2%±3.8% vs 8.8%±1.2%（t=26.44，P＜0.01）；荧光强度：4263±273 vs 1444±212（t=19.27，P＜0.01）；木瓜蛋白酶处理后抗原阳性率：62.8%±3.6% vs 8.8%±1.5%（t=21.38，P＜0.01）；荧光强度：4389±284 vs 1458±230（t=17.49，P＜0.01）。进一步通过流式细胞术分析发现，ZZAP的处理可显著减弱红细胞表面JMH抗原的抗原性（抗原阳性率：62.2%±4.4% vs 48.2%±4.1%（t=14.87，P＜0.01）；荧光强度：4106±263 vs 2063±175（t=17.49，P＜0.01）.
结论: 2-Me处理可减弱人红细胞表面JMH抗原的抗原性；DTT处理后可增强人红细胞表面JMH抗原的抗原性；菠萝蛋白酶与木瓜蛋白酶的处理均可大幅度减弱人红细胞表面JMH抗原的抗原性；ZZAP可降低人红细胞表面JMH抗原的抗原性.